CHAPTER 6 IN-VITRO PHARMACOLOGICAL STUDIES
|
|
- Iris Ursula Miller
- 6 years ago
- Views:
Transcription
1 95 CHAPTER 6 IN-VITRO PHARMACOLOGICAL STUDIES The biological experiments like in-vitro and in-vivo screening models for various diseases may be applied to discover new molecules. These contemporary techniques include bio evaluation, clinical trials, toxicological studies etc. The use of disease induced animal models or the genetically transformed disease models are in use to validate the claims of these medicines, but ethical issues and cost effectiveness of these models have discouraged their common use. Besides, these disease induced models very often get reversed to the normal condition in due course of time as a part of natural healing, which also makes it difficult in interpreting the result of a test drug. Therefore, in-vitro mechanism based screening of herbal medicine is mandatory in the initial phases of plant drug research before taking them to in-vivo study to evaluate their efficacy. Special statistical tools must be used to evaluate their efficacy of the collected data, otherwise it may be difficult to reach any conclusion or to frame a hypothesis [50]. Symplocos cochinchinensis (Lour.) is traditionally used to treat various diseases. Since, no literature is available on the biological effects of this plant, the present study was carried out to screen the plant extracts for various in-vitro pharmacological studies like in-vitro cytotoxic activity, in-vitro anti-inflammatory activity and in-vitro anti-snake venom activity for n-hexane, chloroform, ethyl acetate and methanol extracts.
2 IN-VITRO CYTOTOXIC ACTIVITY Introduction Cancer continues to represent the largest cause of mortality in the world and claims over 6 million lives every year [51]. Drug development programmes involve preclinical screening of a vast number of chemicals for their specific and non specific cytotoxicity against many types of cells. Use of in-vitro assay system for the screening of potential anticancer drug has been a common practice almost since the beginning of cancer therapy in The National Cancer Institute now routinely measure the growth inhibitory properties of every compound under test against a panel of human tumor cell lines which are representative of major human tumor types. There are number of advantages in in-vitro test using cell cultures which includes analysis of species specificity, feasibility of using only small amount of test substance and facility to do mechanistic studies. A novel anticancer drug should possess cytotoxicity at low concentration against cancerous cell lines and should be safe against normal cell lines even at higher concentration [52]. The direct antitumor activity of the plant extracts can be tested under in-vitro conditions using cultured or fresh preparation of various cancer cell types. Once the activity has been detected, the study has to be followed up vigorously to establish therapeutic efficacy and safety [53] Materials and Methods Method : MTT Assay methanol extracts Plant materials used: n-hexane, chloroform, ethyl acetate and
3 97 Cell culture Human breast cancer- MDA-MB-231, Colon cancer-sw 620, Liver cancer Hep G 2 were obtained from National centre for cell science (Pune, India). Chemicals Dubelcco s Modified Eagle s Minimum Essential Medium (DMEM), Foetal Calf Serum (FCS), trypsin (0.25%) were from D. Dutscher (Brumath, France). 3- (4, 5 dimethyl thiazo l -2-yl)- 2,5 diphenyl tetrazolium bromide (MTT) and dimethylsulfoxide (DMSO). Principle This colorimetric assay is based on the capacity of mitochondria succinate dehydrogenase enzymes in living cells to reduce the yellow water soluble substrate 3- (4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) into an insoluble, blue colored formazan product which is measured spectrophotometrically. Only viable cells with active mitochondria reduce significant amounts of MTT [54] since reduction of MTT can only occur in metabolically active cells, the level of activity is a measure of the viability of the cells[55]. Cell lines Human breast cancer- MDA-MB-231, Colon cancer-sw 620, Liver cancer Hep G 2 were obtained from National centre for cell science (Pune, India). Stock culture of these cell lines were cultured in RPMI or DMEM supplemented with 10% inactivated newborn calf serum, Penicillin (100 IU/ml), Streptomycin (100ìg/ml) and Amphotericin (5 ìg/ml) under humidified atmosphere of 5% CO 2 at 37 0 C until confluent. The cells were
4 98 dissociated in 0.2% trypsin and 0.02% EDTA in phosphate buffer saline solution. The stock culture was grown in 25cm tissue culture flasks and cytotoxicity experiments were carried out in 96 well microtiter plates (Tarsons India, Kolkata, India). Procedure Cell lines in the exponential growth phase were washed, trypsinized and suspended in complete culture media. Cells were plated at 10,000 cells/well in 96 well microtiter plates and incubated for 24hrs during which a partial monolayer was formed. They were then exposed to various concentrations of the extract (1 100mcg/ml). Control wells received only maintenance medium. The plates were incubated at 37 C in a humidified incubator with 5% CO 2 for a period of 72 hrs and cells were periodically checked for granularity, shrinkage and swelling. After 72 hrs, the sample solution in wells was flicked off and 50 l of MTT dye was added to each well. The plates were gently shaken and incubated for 4 hrs at 37 0 C in 5% CO 2 incubator. The supernatant was removed and 50 l of propanol was added. The plates were gently shaken to solubilise the formed formazan. The absorbance was measured at 540nm [56]. The percentage growth inhibition was calculated using the following formula, Mean OD of individual test group Percentage growth inhibition = 100 (6.2) Mean OD of control group Values of absorbance were converted into percentage of residual viability. Usually, inhibition concentration 50% (GI 50 ) is chosen as the best biological marker of cytotoxicity. The GI 50 value represents the concentration of the tested extracts that caused 50% of cell inhibition.
5 Results The in-vitro cytotoxic activity of n-hexane, chloroform, ethyl acetate and methanol extracts was given in Table 6.1 Table 6.1 The in-vitro cytotoxic activity of various extracts of leaves of Symplocos cochinchinensis (Lour.) MDA-MB-231 ( g/ml) SW 620 ( g/ml) Hep G 2 ( g/ml) Extracts GI 50 TGI GI 50 TGI GI 50 TGI N-Hexane >100 >100 >100 >100 >100 >100 Chloroform >100 >100 >100 >100 >100 >100 Ethyl acetate 50 > >100 Methanol Average of 3 determinations, 3 replicates GI 50 - Drug concentration inhibiting 50% cellular growth TGI- Total cellular growth inhibition Discussion In in-vitro cytotoxic study of both ethyl acetate and methanol extracts showed significant activity against the three human cancer cell lines namely Human breast cancer- MDA-MB-231, Colon cancer-sw 620, Liver cancer Hep G 2. The n-hexane and chloroform extracts did not show any activity. Methanol extract showed greater cytotoxic effect against colon cancer cell lines SW 620 and HepG2 (liver cancer cell) with a GI 50 value below 20 g/ml when compared to ethyl acetate extract.
6 IN-VITRO ANTI-INFLAMMATORY ACTIVITY Introduction The inflammatory response involves a complex array of enzyme activation, mediator release, cell migration, tissue breakdown and repair[57] which are aimed at host defense and usually activated in most disease condition. Currently much interest have been paid in the searching of medicinal plants with anti-inflammatory activity which may lead to the discovery of new therapeutic agent that is not only used to suppress the inflammation but also used in diverse disease conditions where the inflammation response is amplifying the disease process. In this work the various extracts of Symplocos cochinchinensis (Lour.) were studied for its in-vitro anti-inflammatory activity Materials and Method Plant material n-hexane, chloroform, ethyl acetate and methanol extracts Method The human red blood cell membrane stabilization method Procedure [58] The blood was collected from healthy human volunteers who have not taken any NSAIDS for 2 weeks prior to the experiment and mixed with equal volume of Alsever s solution(2% dextrose, 0.8% sodium citrate, 0.5% citric acid and 0.42% NaCl) and centrifuged at 3,000 rpm. The packed cells were washed with isosaline and a 10% suspension was made. Various concentrations of extracts were prepared (, and 1000 mcg/ml) using distilled water and to each concentration 1 ml of phosphate buffer, 2 ml hyposaline and 0.5 ml of HRBC suspension were added. It was incubated at 37 0 C for 30 min and centrifuged at 3,000 rpm for 20 min. The haemoglobin
7 101 content of the supernatant solution was estimated spectrophotometrically at 560 nm. Diclofenac (50 mcg/ml) was used as reference standard and a control was prepared omitting the extracts. The percentage inhibition of lysis was calculated Results The results of in-vitro anti-inflammatory activity of extracts of Symplocos cochinchinensis (Lour.) by HRBC membrane stabilization method was tabulated in Table 6.2 and Figure 6.1. Table 6.2 In-vitro anti-inflammatory activity of extracts of Symplocos cochinchinensis(lour.) by HRBC membrane stabilization method S.No Treatment Conc(mcg/ml) Absorbance(540nm) %Inhibition 1. Control ± n-hexane Chloroform Ethyl acetate Methanol ± ± 0.01 ** 0.38 ± ± *** 0.33 ± *** 0.35 ± *** 0.25 ± *** 0.28 ± *** 0.29 ± *** 0.15 ± *** 0.19 ± *** 0.20 ± Diclofenac ± *** 73.9 Values are expressed as mean ± SEM. n=5 ***P<0.001 **P< 0.01 compared to control group
8 102 (%) Inhibition n-hexane Chloroform Ethyl Acetate Methanol Diclofenac Figure 6.1 Effect of various extracts on HRBC membrane stabilization method Discussion Among all the extracts tested the methanolic extract showed significant anti-inflammatory activity in a concentration dependent manner. Methanol extract at a concentration of 1000 mcg/ml showed 69.9% protection of HRBC in hypotonic solution. All the results were compared with standard Diclofenac which showed 73.9% protection. It is well known that the vitality of the cells depends on the integrity of their membranes. Exposure of red blood cell to hypotonic medium results in lysis of its membrane accompanied by haemolysis and oxidation of haemoglobin [59,60]. The haemolytic effect of hypotonic solution is related to excessive accumulation of fluid within the cell resulting in the rupturing of its membrane. It is therefore expected that compounds with membrane-stabilizing properties, should offer significant protection of cell membrane against injurious substances. The lysosomal enzymes released during inflammation produces a variety of disorders. The extracellular activity
9 103 of these enzymes is said to be related to acute or chronic inflammation. There is increasing evidence that lysosomal enzymes play an important role in the development of acute and chronic inflammation [61]. Most of the anti-inflammatory drugs exert their beneficial effects by inhibiting either release of these enzymes or by stabilizing lysosomal membrane, which is one of the major event responsible for the inflammatory process [62]. The extracts exhibited membrane stabilization effect by inhibiting hypotonicity induced lysis of erythrocyte membrane. The erythrocyte membrane is analogous to the lysosomal membrane [63] and its stabilization implies that the extract may as well stabilize the lysosomal membranes. Stabilization of lysosomal membrane is important in limiting the inflammatory response by preventing the release of lysosomal constituents of activated neutrophil such as bactericidal enzymes and proteases, which cause further tissue inflammation and damage upon extra cellular release [64]. Some of the NSAIDs are known to possess membrane stabilization properties which may contribute to the potency of their anti-inflammatory effect. Though the exact mechanism of the membrane stabilization by the extract is not known yet, hypotonicity-induced haemolysis may arise from shrinkage of the cells due to osmotic loss of intracellular electrolyte and fluid components. From this result, it is suggested that anti-inflammatory activity observed in this study, may be due to the ability of the extracts to interfere with the early phase of inflammatory reactions, which may stimulate or enhance the efflux of these intracellular components [65] there by exhibits the anti-inflammatory activity.
10 IN-VITRO ANTI-SNAKE VENOM ACTIVITY Introduction Snake envenoming is a major public health issue in the rural tropics with large numbers of envenoming and deaths. In India there are about 216 different snake species of which 53 species are reported to be poisonous. Due to high cost and unavailability of anti venoms, the use of plant remedies for the treatment is the most common practice in India. Extracts from plants have been used among traditional healers, especially in tropical areas for snakebite for a long time [66]. Several medicinal plants, which appear in old drug recipes or which have been passed on by oral tradition are believed to be snakebite antidotes [67]. In almost any part of the world, where venomous snakes occur, numerous plant species are used as folk medicine to treat snakebite. However many of the reported studies lacks detailed scientific investigation, which is needed in the development of medicinal agents from plant sources. Symplocos cochinchinensis (Lour.) is traditionally used in the treatment of snake bites. Since, the anti-snake venom properties of this plant is not scientifically proved, the present study has been undertaken to investigate the in- vitro anti-snake venom activity of various extracts of the leaves Materials and Methods Plant extract n-hexane, chloroform, ethyl acetate and methanol extracts Snake venom Lyophilized snake venom of Russell s viper (Daboia russelli) was obtained from CSIR Centre for Biochemical s, New Delhi and preserved at
11 C. The snake venom was dissolved in 0.9%(w/v) saline, centrifuged and the supernatant was used whenever required. The venom concentration was expressed in terms of dry weight (mg/ml) of the stock venom. Method - Inhibition of in-vitro Human Red Blood Corpuscles lysis method Procedure n-hexane, chloroform, ethyl acetate and methanolic extracts at a concentration of 100,, and 1000mcg/ml of the leaves of Symplocos cochichinensis (Lour.) were used for the study. The hypo saline induced haemolysis was evaluated in-vitro by the method of Roelofsen et al and Balu et al [68,69]. This method was modified in the present study by venominduced haemolysis. Blood was collected from healthy human volunteers by vein puncture and heparin was used as an anti coagulant. The collected blood was washed three times with physiological saline solution to make a stock solution of 100mcg/ml. 1 ml of the venom, 1 ml phosphate buffer(ph 7.4) and 1 ml of 1% HRBC were taken in various tubes. Different concentrations of n-hexane, chloroform, ethyl acetate and methanolic extracts of the leaves (100,, and 1000mcg/ml) were added. The extracts were prepared by using saline and carboxy methyl cellulose (CMC) as suspending agent and the control was prepared omitting the extracts. The anti-snake venom serum(100mcg/ml) was used as standard. These mixtures were incubated at 37 0 C for 30 min and then centrifuged at 1000 rpm for 3 min. The absorbance of the supernatant was measured at 540 nm using spectrophotometer. The percentage inhibition of haemolysis was calculated by using the following formula A (Control) A (test) % inhibition = X 100 (6.2) A (Control)
12 Results The in-vitro anti-snake venom activity of various extracts of the leaves of Symplocos cochinchinensis (Lour.) were studied against Russell s viper venom. The results were tabulated in the Table 6.3 and Figure 6.2. Table 6.3 In-vitro anti-snake venom activity (Inhibition of human red blood cell lysis method) S.No Treatment Conc.(mcg/ml) % inhibition 1. Contorl Snake venom antiserum n-hexane extract Chloroform extract Ethyl acetate extract Methanol extract
13 (%) Inhibition n-hexane Chloroform Ethyl Acetate Methanol Snake venom antiserum Figure 6.2 In-vitro anti-snake venom activity Discussion In-vitro anti-snake venom activity was carried out by prevention of HRBC membrane lysis method. Haemolysis is one of the major causes of snake venomation. Most of the snake venom contains phospholipase and haemolysin[70], which acts on membrane associated phospholipids to liberate lysolecithin. Lysolecitihin acts on the membrane of HRBC causing haemolysis [71]. The extracts inhibit the haemolysis induced by Russell s viper venom in a concentration dependent manner. The percentage inhibition of haemolysis activity was found to be significant in methanol extract at a concentration of 1000mcg/ml. It showed 69.5% protection against venom induced haemolysis which may be due to the stabilization of the protein in the membrane of HRBC [72]. Hence, it may be suggested that the extract may interact with Russell s viper venom and stabilize the protein in the membrane.
IN VITRO ANTICANCER ACTIVITY OF FLOWER EXTRACTS OF COUROUPITA GUIANENSIS
CHAPTER 3 IN VITRO ANTICANCER ACTIVITY OF FLOWER EXTRACTS OF COUROUPITA GUIANENSIS 3. INTRODUCTION Plants are the basic source of knowledge of modern medicine. Almost all the parts of the plant, namely
More informationInternational Journal Of Recent Scientific Research
International Journal Of Recent Scientific Research ISSN: 0976-3031 Volume: 7(5) May -2016 NEW ERA ABOUT TREATMENT - EVOLUATION OF ANTICANCER PROPERTIES OF EVERGREEN MEDICINAL PLANT ANNONA MURICATA (GRAVIOLA)
More information8. CHAPTER IV. ANTICANCER ACTIVITY OF BIOSYNTHESIZED SILVER NANOPARTICLES
8. CHAPTER IV. ANTICANCER ACTIVITY OF BIOSYNTHESIZED SILVER NANOPARTICLES 8.1. Introduction Nanobiotechnology, an emerging field of nanoscience, utilizes nanobased-systems for various biomedical applications.
More informationJournal of Chemical and Pharmaceutical Research
Available on line www.jocpr.com Journal of Chemical and Pharmaceutical Research ISSN No: 0975-7384 CODEN(USA): JCPRC5 J. Chem. Pharm. Res., 2011, 3(2):144-149 In vitro hepatoprotective activity of ethanolic
More informationIn-vitro assay for Cytotoxicity activity in ethonolic extract of fruit rind of Couropita Guianensis aubl
ISSN: 2319-7706 Volume 3 Number 10 (2014) pp. 169-176 http://www.ijcmas.com Original Research Article In-vitro assay for Cytotoxicity activity in ethonolic extract of fruit rind of Couropita Guianensis
More informationAnti-cancer activity of Aya Thambira Chendooram (ATC) in in-vitro cell line against Breast Carcinoma
International Journal of Advanced Research in Biological Sciences ISSN: 2348-8069 www.ijarbs.com DOI: 10.22192/ijarbs Coden: IJARQG(USA) Volume 5, Issue 1-2018 Research Article DOI: http://dx.doi.org/10.22192/ijarbs.2018.05.01.010
More informationSupplementary Files S1 Isolation of Monocytes S2 Haemolysis study Reagents Procedure S3 Cytotoxicity studies Trypan blue dye exclusion method
Electronic Supplementary Material (ESI) for RSC Advances. This journal is The Royal Society of Chemistry 2014 Supplementary Files S1 Isolation of Monocytes A 3 ml volume of Histopaque 1083 solution was
More informationJournal of Chemical and Pharmaceutical Research, 2013, 5(5): Research Article. Anticancer properties of Cissus quandrangularis
Available online wwwjocprcom Journal of Chemical and Pharmaceutical Research, 13, 5(5):135-139 Research Article ISSN : 975-7384 CODEN(USA) : JCPRC5 Anticancer properties of Cissus quandrangularis Aayush
More informationAsian Journal of Research in Pharmaceutical Sciences and Biotechnology
Research Article ISSN: 2349 7114 Asian Journal of Research in Pharmaceutical Sciences and Biotechnology Journal home page: www.ajrpsb.com IN VITRO ANTI-TUMOUR ACTIVITY OF VITEX LEUCOXYLON LINN USING MTT
More informationJournal of Chemical and Pharmaceutical Research, 2017, 9(12): Research Article
Available online www.jocpr.com Journal of Chemical and Pharmaceutical Research, 2017, 9(12):30-34 Research Article ISSN : 0975-7384 CODEN(USA) : JCPRC5 Evaluation of Anticancer Activity of Alphonsea Sclerocarpa
More informationCHAPTER 4 IMMUNOLOGICAL TECHNIQUES
CHAPTER 4 IMMUNOLOGICAL TECHNIQUES Nitroblue Tetrazolium Chloride (NBT) Reduction test NBT reduction test was evaluated by employing the method described by Hudson and Hay,1989 based upon principle that
More informationMTS assay in A549 cells
Project: VIGO MTS assay in A549 cells Detection of cell viability/activity AUTHORED BY: DATE: Cordula Hirsch 20.01.2014 REVIEWED BY: DATE: Harald Krug 10.04.2014 APPROVED BY: DATE: DOCUMENT HISTORY Effective
More informationEvaluation of Biological Activity (In-Vitro) of Some 2-Phenyl Oxazoline Derivatives
Human Journals Research Article April 2016 Vol.:6, Issue:1 All rights are reserved by V.Niraimathi et al. Evaluation of Biological Activity (In-Vitro) of Some 2-Phenyl Oxazoline Derivatives Keywords: Antimicrobial
More informationCHAPTER 11 INVITRO ANTI-OXIDANT ACTIVITY
152 CHAPTER 11 INVITRO ANTI-OXIDANT ACTIVITY 11.1 INTRODUCTION In living systems, free radicals are generated as part of the body's normal metabolic process and the free radical chain reactions are usually
More informationIn Vitro Antioxidant and Cytotoxic Analysis of Boerhaavia diffusa Linn.
Ethnobotanical Leaflets 13: 263-68. 2009. In Vitro Antioxidant and Cytotoxic Analysis of Boerhaavia diffusa Linn. Teepica Priya Darsini D* 1, J.M. Sasikumar 1 and Kulandhaivel M. 2 1 Department of Industrial
More informationSUPPLEMENTARY MATERIAL
SUPPLEMENTARY MATERIAL Acetone and methanol fruit extracts of Terminalia paniculata inhibit HIV-1 infection in vitro Ankita Durge a, Pratiksha Jadaun a, Ashish Wadhwani a, Ashish A. Chinchansure b, Madhukar
More informationAnti Cancer Activity of Moringa Oleifera (Flowers) Against Human Liver Cancer
4 IJISET - International Journal of Innovative Science, Engineering & Technology, Vol. 4 Issue 6, June 17 Anti Cancer Activity of Moringa Oleifera (Flowers) Against Human Liver Cancer 1* 3 4 A. Rajeshkanna,
More informationMTS assay in THP-1 cells
Project: VIGO MTS assay in THP-1 cells Detection of cell viability/activity AUTHORED BY: DATE: Cordula Hirsch 20.01.2014 REVIEWED BY: DATE: Harald Krug 10.04.2014 APPROVED BY: DATE: DOCUMENT HISTORY Effective
More informationScholars Research Library J. Nat. Prod. Plant Resour., 2017, 7(2): (
Available online www.scholarsresearchlibrary.com Scholars Research Library J. Nat. Prod. Plant Resour., 2017, 7(2): 74-78 (http://scholarsresearchlibrary.com/archive.html) Evaluation of Anti-Inflammatory
More informationSupplementary Information. Sonorensin: A new bacteriocin with potential of an anti-biofilm agent and a food
Supplementary Information Sonorensin: A new bacteriocin with potential of an anti-biofilm agent and a food biopreservative Lipsy Chopra, Gurdeep Singh, Kautilya Kumar Jena and Debendra K. Sahoo* Biochemical
More informationAppendix A: Preparation of Media and Chemicals. Malt Extract Agar (MEA) weighing g was dissolved in 400 ml of distilled water
Appendix A: Preparation of Media and Chemicals Preparation of Malt Extract Agar (MEA) Malt Extract Agar (MEA) weighing 13.44 g was dissolved in 400 ml of distilled water in an Erlenmeyer flask using a
More information3. RESULTS AND DISCUSSION
3. RESULTS AND DISCUSSION 3.1. PHYTOCHEMICAL INVESTIGATIONS OF F.RACEMOSA BARK The course powder of Ficus racemosa was sequentially extracted by Pet. Ether, Ethyl acetate along with Alcohol by Soxhlet
More informationInvestigations on its antioxidative and anti-inflammatory potential
- 1 - CITROZINE Investigations on its antioxidative and CITROFRESH SUPERCONCENTRATE anti-inflammatory potential Investigator and responsible for the correctness of the test protocol, results, conclusions
More informationSeparation of Plasma and Serum and Their Proteins from Whole Blood
Separation of Plasma and Serum and Their Proteins from Whole Blood BCH 471 [Practical] BLOOD COMPOSITION Other names to blood cells Red blood cells (erythrocytes) White blood cells (leukocytes) Platelets
More informationB16-F10 (Mus musculus skin melanoma), NCI-H460 (human non-small cell lung cancer
Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 2017 Experimental Methods Cell culture B16-F10 (Mus musculus skin melanoma), NCI-H460 (human non-small
More informationProtease Assay. (Cat. # ) think proteins! think G-Biosciences
389PR 01 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Protease Assay (Cat. # 786 028) think proteins! think G-Biosciences www.gbiosciences.com
More informationLankenau Institute for Medical Research Annual Progress Report: 2011 Formula Grant
Lankenau Institute for Medical Research Annual Progress Report: Formula Grant Reporting Period January, June, Formula Grant Overview The Lankenau Institute for Medical Research received $6,99 in formula
More informationPreparation of stock solution and reagents for DPPH assay
A1 Preparation of stock solution and reagents for DPPH assay i. Plant sample (Stock solution) A stock solution of 20 mg/ml of each extract was prepared and wrapped in aluminium foil. The crude methanol
More informationWork-flow: protein sample preparation Precipitation methods Removal of interfering substances Specific examples:
Dr. Sanjeeva Srivastava IIT Bombay Work-flow: protein sample preparation Precipitation methods Removal of interfering substances Specific examples: Sample preparation for serum proteome analysis Sample
More informationBIOL 347L Laboratory Three
Introduction BIOL 347L Laboratory Three Osmosis in potato and carrot samples Osmosis is the movement of water molecules through a selectively permeable membrane into a region of higher solute concentration,
More informationLow Cell Binding Property of LIPIDURE -COAT
Technical Note_1 ver.1 Low Cell Binding Property of LIPIDURE -COAT 1. LIPIDURE -COAT MULTI DISH A-6MD (Cat. No. 51011617) 2. Cell; NIH 3T3 (Fibroblast, mouse) 1. 10 %CS-DMEM; DMEM (Dulbecco's Modified
More informationAnticancer Properties of Phytochemicals Present in Indian Medicinal Plants.
Anticancer Properties of Phytochemicals Present in Indian Medicinal Plants. Dr.Prathiba H.D* Department Of Biochemistry, Central College Campus, Bangalore University. Bangalore(Karnataka), INDIA. ABSTRACT
More informationTEST REPORT & SPECIFIC INFORMATION
Page 1 (5) Dartsch Scientific GmbHAuf der Voßhardt 25 D-49419 Wagenfeld Firma LuKo Pharm GmbH Mayrwiesstrasse 25-27 A-5300 Hallwang Auf der Voßhardt 25 D-49419 Wagenfeld, Germany Fon: +49 5444 980 1322
More informationEffect of Organically Grown Curcuma longa (Turmeric) on Leukemic and MCF-7 Cell Lines
ISSN: 2319-7706 Special Issue-2 (May-2015) pp. 182-186 http://www.ijcmas.com Original Research Article Effect of Organically Grown Curcuma longa (Turmeric) on Leukemic and MCF-7 Cell Lines Priyanka Argade
More informationIn-vitro Cytotoxic Activity of Indianthus virgatus (Roxb.) Suksathan and Borchs. On A549, A431, CaCo2, U87 and L929 Cell Lines
Pharmacogn J. 2018; 10(6):1216-1220 A Multifaceted Journal in the field of Natural Products and Pharmacognosy www.phcogj.com www.journalonweb.com/pj www.phcog.net Original Article In-vitro Cytotoxic Activity
More informationAZO-XYLAN (BIRCHWOOD)
ASSAY OF endo-1,4-ß-xylanase using AZO-XYLAN (BIRCHWOOD) S-AXBP S-AXBL 10/07 Megazyme International Ireland 2007 PRINCIPLE: This assay procedure is specific for endo-1,4-ß-d-xylanase activity. On incubation
More informationBIOCHEMISTRY OF BLOOD
BCH 471 BIOCHEMISTRY OF BLOOD Amal Alamri Experiment 1 Separation of Plasma and Serum from Whole Blood Whole Blood It is living tissue that circulates through the heart, arteries, veins, and capillaries
More informationAnti-Aging Activity Of Cucurbita moschata Ethanolic Extract Towards NIH3T3 Fibroblast Cells Induced By Doxorubicin
Indonesian Journal of Cancer Chemoprevention, 2016, 7(2): 49-53 ISSN: 2088 0197 Anti-Aging Activity Of Cucurbita moschata Ethanolic Extract Towards NIH3T3 Fibroblast Cells Induced By Doxorubicin Laeli
More informationPFK Activity Assay Kit (Colorimetric)
PFK Activity Assay Kit (Colorimetric) Catalog Number KA3761 100 assays Version: 02 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 General Information...
More informationEPIGENTEK. EpiQuik Global Histone H3 Acetylation Assay Kit. Base Catalog # P-4008 PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE
EpiQuik Global Histone H3 Acetylation Assay Kit Base Catalog # PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE The EpiQuik Global Histone H3 Acetylation Assay Kit is suitable for specifically measuring global
More informationAnti-Aging Skin Care Regimen. Phyto-Stem Cell: Advanced Plant Cell Culture Technology by Bio-FD&C
Anti-Aging Skin Care Regimen Phyto-Stem Cell: Advanced Plant Cell Culture Technology by Bio-FD&C 01 Plant Cell Culture Technology Highly expression of phytochemicals from plant cell culturing process by
More informationHT Glutathione Assay Kit
Instructions For Research Use Only. Not For Use In Diagnostic Procedures HT Glutathione Assay Kit Colorimetric assay for total, reduced and oxidized glutathione. Sufficient reagents for tests. Table of
More informationEPIGENTEK. EpiQuik Global Histone H4 Acetylation Assay Kit. Base Catalog # P-4009 PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE
EpiQuik Global Histone H4 Acetylation Assay Kit Base Catalog # PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE The EpiQuik Global Histone H4 Acetylation Assay Kit is suitable for specifically measuring global
More informationCoQ10(Coenzyme Q10) ELISA Kit
CoQ10(Coenzyme Q10) ELISA Kit Catalogue No.: EU0196 Size: 48T/96T Reactivity: Universal Detection Range: 0.781-50ng/ml Sensitivity:
More informationCytotoxicity Effect of Zingiber officinale, Curcuma aeruginosa and Curcuma xanthorhiza Extracts on Adipose-Derived Stem Cells (ADSCs)
Cytotoxicity Effect of Zingiber officinale, Curcuma aeruginosa and Curcuma xanthorhiza Extracts on Adipose-Derived Stem Cells (ADSCs) Rilianawati 1, Mutia Hardhiyuna 1, Angelia Yulita 1, Ago Halim 2 1
More informationTitle Revision n date
A. THIN LAYER CHROMATOGRAPHIC TECHNIQUE (TLC) 1. SCOPE The method describes the identification of hydrocortisone acetate, dexamethasone, betamethasone, betamethasone 17-valerate and triamcinolone acetonide
More informationMATERIAL AND METHODS
MATERIAL AND METHODS Material and Methods Glucose induced cataract was chosen as a model for the present study. A total of 210 fresh goat lenses were analyzed. Sample Collection: Goat eyeballs were obtained
More informationHCC1937 is the HCC1937-pcDNA3 cell line, which was derived from a breast cancer with a mutation
SUPPLEMENTARY INFORMATION Materials and Methods Human cell lines and culture conditions HCC1937 is the HCC1937-pcDNA3 cell line, which was derived from a breast cancer with a mutation in exon 20 of BRCA1
More informationBIL 256 Cell and Molecular Biology Lab Spring, Tissue-Specific Isoenzymes
BIL 256 Cell and Molecular Biology Lab Spring, 2007 Background Information Tissue-Specific Isoenzymes A. BIOCHEMISTRY The basic pattern of glucose oxidation is outlined in Figure 3-1. Glucose is split
More informationANTI-INFLAMMATORY ACTIVITY OF MORINDA CITRIFOLIA EXTRACT-AN INVITRO STUDY
ISSN: 0975-766X CODEN: IJPTFI Available Online through Research Article www.ijptonline.com ANTI-INFLAMMATORY ACTIVITY OF MORINDA CITRIFOLIA EXTRACT-AN INVITRO STUDY 1 A.Trishala, 1 T.Lakshmi 1 BDS, Saveetha
More informationSTORE AT 4 o C Version 3
Instruction Manual Advanced Protein Assay Reagent (Cat. # ADV01) ORDERING INFORMATION To order by phone: (303) - 322-2254 To order by Fax: (303) - 322-2257 To order by e-mail: cserve@cytoskeleton.com Technical
More informationTotal Histone H3 Acetylation Detection Fast Kit (Colorimetric)
Total Histone H3 Acetylation Detection Fast Kit (Colorimetric) Catalog Number KA1538 48 assays Version: 02 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use...
More informationRecombinant Trypsin, Animal Origin Free
Recombinant Trypsin, Animal Origin Free PRODUCT INFORMATION: BioGenomics r-trypsin powder is ready to use, animal origin free optimized for cell culture applications. It is derived by r-dna technology.
More informationGlobal Histone H3 Acetylation Assay Kit
Global Histone H3 Acetylation Assay Kit Catalog Number KA0633 96 assays Version: 06 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use... 3 Background... 3 Principle
More informationIN VITRO HORMESIS EFFECTS OF SODIUM FLUORIDE ON KIDNEY CELLS OF THREE-DAY-OLD MALE RATS
292 Tang, An, Du, Zhang, Zhou 292 IN VITRO HORMESIS EFFECTS OF SODIUM FLUORIDE ON KIDNEY CELLS OF THREE-DAY-OLD MALE RATS Qin-qing Tang, a Xiao-jing An, a Jun Du, a Zheng-xiang Zhang, a Xiao-jun Zhou a
More informationSupporting Information Nitric oxide releasing photoresponsive nanohybrids as excellent therapeutic agent for cervical cancer cell lines
upporting Information itric oxide releasing photoresponsive nanohybrids as excellent therapeutic agent for cervical cancer cell lines Priya udhesh a, Kaviyarasan Tamilarasan a, Palaniappan Arumugam a and
More informationAPPENDIX Heparin 2 mg heparin was dissolved in 0.9 % NaCl (10 ml). 200 µl of heparin was added to each 1 ml of blood to prevent coagulation.
APPENDIX 1 Preparation of reagents 1.1. Preparation of dosing solution Nonylphenol 15 mg of Nonylphenol was dissolved in olive oil (10 ml) and used as stock solution. The stock solution was serially diluted
More informationCELLULASE from PENICILLIUM FUNICULOSUM
CELLULASE from PENICILLIUM FUNICULOSUM Prepared at the 55th JECFA (2000) and published in FNP 52 Add 8 (2000), superseding tentative specifications prepared at the 31st JECFA (1987) and published in FNP
More informationTaraxacum officinale Herb as an Antiinflammatory
American Journal Advanced Drug Delivery www.ajadd.co.uk Taraxacum ficinale Herb as an Antiinflammatory Medicine M. Amin Mir* 1, S.S. Sawhney 1 and Manmohan Singh Jassal 2 1 R & D Division, Uttaranchal
More informationEXPERIMENT 13: Isolation and Characterization of Erythrocyte
EXPERIMENT 13: Isolation and Characterization of Erythrocyte Day 1: Isolation of Erythrocyte Steps 1 through 6 of the Switzer & Garrity protocol (pages 220-221) have been performed by the TA. We will be
More informationApoptosis Mediated Cytotoxicity of Curcumin Analogues PGV-0 and PGV-1 in WiDr Cell Line
Apoptosis Mediated Cytotoxicity of Curcumin Analogues PGV-0 and PGV-1 in WiDr Cell Line Endah Puji Septisetyani, Muthi Ikawati, Barinta Widaryanti and Edy Meiyanto* ) Cancer Chemoprevention Research Center,
More informationab Adipogenesis Assay Kit (Cell-Based)
ab133102 Adipogenesis Assay Kit (Cell-Based) Instructions for Use For the study of induction and inhibition of adipogenesis in adherent cells. This product is for research use only and is not intended
More information20X Buffer (Tube1) 96-well microplate (12 strips) 1
PROTOCOL MitoProfile Rapid Microplate Assay Kit for PDH Activity and Quantity (Combines Kit MSP18 & MSP19) 1850 Millrace Drive, Suite 3A Eugene, Oregon 97403 MSP20 Rev.1 DESCRIPTION MitoProfile Rapid Microplate
More information21 Virginiamycin OH O. For chickens (except for broilers) broilers. Added amount 5~15 5~15 10~20 10~20
21 Virginiamycin H H H H H H Virginiamycin M 1 C 28 H 35 3 7 MW: 525.6 CAS o.: 21411-53-0 Virginiamycin S 1 C 43 H 49 7 10 MW: 823.9 CAS o.: 23152-29-6 [Summary of virginiamycin] Virginiamycin (VM) is
More informationEpiQuik Total Histone H3 Acetylation Detection Fast Kit (Colorimetric)
EpiQuik Total Histone H3 Acetylation Detection Fast Kit (Colorimetric) Base Catalog # PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE The EpiQuik Total Histone H3 Acetylation Detection Fast Kit (Colorimetric)
More informationEFFICACY OF APAMARGA KSHARA IN CERVICAL CANCER CELL LINES
INTERNATIONAL AYURVEDIC MEDICAL JOURNAL International Ayurvedic Medical Journal, (ISSN: 2320 5091) (July, 2017) 5(7) EFFICACY OF APAMARGA KSHARA IN CERVICAL CANCER CELL LINES Mrudula K. S 1, Mamatha K.V
More informationProcine sphingomyelin ELISA Kit
Procine sphingomyelin ELISA Kit For the quantitative in vitro determination of Procine sphingomyelin concentrations in serum - plasma - celiac fluid - tissue homogenate - body fluid FOR LABORATORY RESEARCH
More informationAntioxidant Activity of the plant Andrographis paniculata (Invitro)
Chapter 4 Antioxidant Activity of the plant Andrographis paniculata (Invitro) 4.1 INTRODUCTION Antioxidants prevents or repairs the cells against reactive oxygen species, reduces damage caused by free
More informationAlkaline Phosphatase Assay Kit
Alkaline Phosphatase Assay Kit Catalog Number KA0817 500 assays Version: 02 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 General Information... 4 Materials
More informationElectronic Supporting Information for
Electronic Supplementary Material (ESI) for RSC Advances. This journal is The Royal Society of Chemistry 2015 Electronic Supporting Information for Rhodamine based Turn-On Fluorescent Probe for Pb(II)
More informationAMYLOGLUCOSIDASE from ASPERGILLUS NIGER, var.
AMYLOGLUCOSIDASE from ASPERGILLUS NIGER, var. SYNONYMS INS No. 1100 Prepared at the 59 th JECFA (2002) and published in FNP 52 Add 10 (2002), superseding tentative specifications prepared at the 55 th
More informationAsian Journal of Research in Biological and Pharmaceutical Sciences Journal home page:
Research Article ISSN: 2349-4492 Asian Journal of Research in Biological and Pharmaceutical Sciences Journal home page: www.ajrbps.com ASSESSMENT OF IN VITRO ANTI INFLAMMATORY ACTIVITY OF AQUEOUS EXTRACT
More informationASSAY OF USING BETA-GLUCAZYME TABLETS
ASSAY OF endo-β-glucanases USING BETA-GLUCAZYME TABLETS T-BGZ 12/12 Note: Changed assay format for malt β-glucanase Megazyme International Ireland 2012 SUBSTRATE: The substrate employed is Azurine-crosslinked
More informationEPIGENTEK. EpiQuik HDAC Activity/Inhibition Assay Kit(Colorimetric) Base Catalog # P-4002 PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE
EpiQuik HDAC Activity/Inhibition Assay Kit(Colorimetric) Base Catalog # PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE The EpiQuik HDAC Activity/Inhibition Assay Kit (Colorimetric) is very suitable for
More informationSensoLyte Generic MMP Assay Kit *Colorimetric*
SensoLyte Generic MMP Assay Kit *Colorimetric* Revision#1.2 Catalog # Kit Size Last updated: May2017 AS-72095 100 Assays (96-well plate) Optimized Performance: This kit is optimized to detect MMP activity
More informationSensoLyte pnpp Alkaline Phosphatase Assay Kit *Colorimetric*
SensoLyte pnpp Alkaline Phosphatase Assay Kit *Colorimetric* Catalog # 72146 Kit Size 500 Assays (96-well plate) Optimized Performance: This kit is optimized to detect alkaline phosphatase activity Enhanced
More informationData Sheet TIGIT / NFAT Reporter - Jurkat Cell Line Catalog #60538
Data Sheet TIGIT / NFAT Reporter - Jurkat Cell Line Catalog #60538 Background: TIGIT is a co-inhibitory receptor that is highly expressed in Natural Killer (NK) cells, activated CD4+, CD8+ and regulatory
More informationWHEN DOES BLOOD HAEMOLYSE? A Temperature Study
Br. J. Anaesth. (1974), 46, 742 WHEN DOES BLOOD HAEMOLYSE? A Temperature Study C. CHALMERS AND W. J. RUSSELL SUMMARY Incubation of blood in vitro for up to 1 hour at temperatures below 45 C C caused no
More informationIN VITRO CELLULAR RESPONSES TO AUTOLOGOUS TUMOR EXTRACT DETECTED BY INHIBITION OF MACROPHAGE MIGRATION*1
[Gann, 66, 167-174; April, 1975] IN VITRO CELLULAR RESPONSES TO AUTOLOGOUS TUMOR EXTRACT DETECTED BY INHIBITION OF MACROPHAGE MIGRATION*1 Tsuyoshi AKIYOSHI, Akira HATA, and Hideo TSUJI Department of Surgery,
More informationHuman Mammary Luminal Epithelial Cells. Manual
Human Mammary Luminal Epithelial Cell Manual INSTRUCTION MANUAL SHIPPING CONDITIONS ZBM0071.00 Human Mammary Luminal Epithelial Cells Orders are delivered via Federal Express courier. All US and Canada
More informationResearch Article. Cell culture study on the cytotoxic effects of Cureit - a novel bio available curcumin-anti cancer effects
Available online www.jocpr.com Journal of Chemical and Pharmaceutical Research, 2014, 6(9):96-100 Research Article ISSN : 0975-7384 CODEN(USA) : JCPRC5 Cell culture study on the cytotoxic effects of Cureit
More informationASSAY OF using CELLAZYME C TABLETS T-CCZ 01/17
www.megazyme.com ASSAY OF endo-cellulase using CELLAZYME C TABLETS T-CCZ 01/17 Megazyme 2017 SUBSTRATE: The substrate employed is azurine-crosslinked HE-cellulose (AZCL-Cellulose). This substrate is prepared
More informationResearch Article GALLIC ACID AND FLAVONOID ACTIVITIES OF AMARANTHUS GANGETICUS
ISSN 2395-3411 Available online at www.ijpacr.com 238 Research Article GALLIC ACID AND FLAVONOID ACTIVITIES OF AMARANTHUS GANGETICUS G. Jyoti Jain 1* and S. Ramachandra Setty 2 1 Department of Pharmacology,
More informationCHAPTER 6 EVALUATION OF SELECTED PLANT EXTRACTS FOR EVALUATION OF SELECTED PLANT EXTRACTS FOR ANTI-ACNE ACTIVITY
CHAPTER 6 EVALUATION OF SELECTED PLANT EXTRACTS FOR School of Science, SVKM s NMIMS University Page 119 6. EVALUATION OF SELECTED PLANT EXTRACTS FOR 6.1 MATERIALS AND METHODS 6.1.1 Antimicrobial assays
More informationab Cathepsin D Human ELISA Kit
ab119586 Cathepsin D Human ELISA Kit Instructions for Use For quantitative detection of Human Cathepsin D in cell culture supernatants, serum and plasma (heparin, EDTA). This product is for research use
More informationScreening of bacteria producing amylase and its immobilization: a selective approach By Debasish Mondal
Screening of bacteria producing amylase and its immobilization: a selective approach By Debasish Mondal Article Summary (In short - What is your article about Just 2 or 3 lines) Category: Bacillus sp produce
More informationProtocol for A-549 VIM RFP (ATCC CCL-185EMT) TGFβ1 EMT Induction and Drug Screening
Protocol for A-549 VIM RFP (ATCC CCL-185EMT) TGFβ1 EMT Induction and Drug Screening Introduction: Vimentin (VIM) intermediate filament (IF) proteins are associated with EMT in lung cancer and its metastatic
More informationThe effect of insulin on chemotherapeutic drug sensitivity in human esophageal and lung cancer cells
The effect of insulin on chemotherapeutic drug sensitivity in human esophageal and lung cancer cells Published in: Natl Med J China, February 10, 2003; Vol 83, No 3, Page 195-197. Authors: JIAO Shun-Chang,
More informationProtocol. G-LISA Cdc42 Activation Assay Biochem Kit : 24 Assays (Absorbance Based) Cat. # BK127-S. cytoskeleton.com. Cytoskeleton, Inc.
The Protein Experts Protocol Cytoskeleton, Inc. V 9.5 G-LISA Cdc42 Activation Assay Biochem Kit : 24 Assays (Absorbance Based) Cat. # BK127-S cytoskeleton.com Phone: (303) 322.2254 Fax: (303) 322.2257
More informationPROTAZYME AK TABLETS
www.megazyme.com ASSAY OF endo-protease using PROTAZYME AK TABLETS T-PRAK 05/16 Megazyme International Ireland 2016 SUBSTRATE: The substrate employed is Azurine-crosslinked casein (AZCL-casein). This substrate
More informationHT Glutathione Assay Kit
IFU0 Rev Status: RELEASED printed //0 ::0 AM by Trevigen Document Control Instructions For Research Use Only. Not For Use In Diagnostic Procedures HT Glutathione Assay Kit Colorimetric assay for total,
More informationPhospholipid Assay Kit
Phospholipid Assay Kit Catalog Number KA1635 100 assays Version: 06 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use... 3 Background... 3 General Information...
More informationLankenau Institute for Medical Research Annual Progress Report: 2011 Formula Grant
Lankenau Institute for Medical Research nnual Progress Report: 2011 Formula Grant Reporting Period July 1, 2012 December 31, 2012 Formula Grant Overview The Lankenau Institute for Medical Research received
More informationTRACP & ALP Assay Kit
Cat. # MK301 For Research Use TRACP & ALP Assay Kit Product Manual Table of Contents I. Description...3 II. III. IV. Introduction...3 Components...4 Materials Required but not Provided...4 V. Storage...4
More informationCELL MEDIATED IMMUNE RESPONSE
CELL MEDIATED IMMUNE RESPONSE Chapter IV - CELL MEDIATED IMMUNE RESPONSE Sujatha, M. 2013. Evaluation of Immunological changes in Fish, Catla catla administered with bacterial pathogen, Aeromonas hydrophila,
More informationab Lipid Peroxidation (MDA) Assay kit (Colorimetric/ Fluorometric)
Version 10b Last updated 19 December 2018 ab118970 Lipid Peroxidation (MDA) Assay kit (Colorimetric/ Fluorometric) For the measurement of Lipid Peroxidation in plasma, cell culture and tissue extracts.
More informationby CM764 in human SK-N-SH neuroblastoma* Hilary Nicholson, Christophe Mesangeau, Christopher R. McCurdy, and Wayne D. Bowen
JPET #228387 Title: Sigma-2 receptors play a role in cellular metabolism: Stimulation of glycolytic hallmarks by CM764 in human SK-N-SH neuroblastoma* Hilary Nicholson, Christophe Mesangeau, Christopher
More informationBlood Urea Nitrogen Enzymatic Kit Manual Catalog #:
Blood Urea Nitrogen Enzymatic Kit Manual Catalog #: 5602-01 TABLE OF CONTENTS GENERAL INFORMATION... 2 Product Description... 2 Procedure Overview... 2 Kit Contents, Storage and Shelf Life... 3 Required
More information