Enzymatic Assay of CREATININASE (EC ) From Pseudomonas species
|
|
- Abel Osborne Underwood
- 5 years ago
- Views:
Transcription
1 PRINCIPLE: Creatinine + H 2 O Creatininase > Creatine Creatine + ATP CPK > Creatine-P + ADP ADP + PEP PK > ATP + Pyruvate Pyruvate + ß-NADH LDH > L-Lactate + ß-NAD Abbreviations used: ATP = Adenosine 5'-Triphosphate CPK = Creatine Phosphokinase Creatine-P = Phosphocreatine ADP = Adenosine 5'-Diphosphate PEP = Phospho(enol)pyruvate PK = Pyruvate Kinase ß-NADH = ß-Nicotinamide Adenine Dinucleotide, Reduced Form LDH = Lactic Dehydrogenase ß-NAD = ß-Nicotinamide Adenine Dinucleotide, Oxidized Form CONDITIONS: T = 25 C, ph = 8.0, A 340nm, Light path = 1 cm METHOD: Continuous Spectrophotometric Rate Determination REAGENTS: A. 100 mm Glycylglycine Buffer, ph 8.0 at 25 C (Prepare 100 ml in deionized water using Gly-Gly, Free Base, Sigma Prod. No G Adjust to ph 8.0 at 25 C with 1 M NaOH.) B. 12 mm ß-Nicotinamide Adenine Dinucleotide, Reduced Form, Solution (ß-NADH) (Dissolve the contents of one 10 mg vial of ß-Nicotinamide Adenine Dinucleotide, Reduced Form, Disodium Salt, Sigma Stock No in the appropriate volume of Reagent A. PREPARE FRESH.) SPCREA01 Page 1 of 4
2 REAGENTS: (continued) C. 16 mm Adenosine 5'-Triphosphate Solution (ATP) (Prepare 5 ml in Reagent A using Adenosine 5'-Triphosphate, Disodium Salt, Sigma Prod. No. A ) D. 20 mm Phospho(enol)pyruvate Solution (PEP) (Prepare 5 ml in Reagent A using Phospho(enol)pyruvate, Tri(cyclohexylammonium) Salt, Sigma Prod. No. P-7252.) E. 440 mm Creatinine Solution (Substrate) (Prepare 5 ml in deionized water using Creatinine, Free Base, Anhydrous, Sigma Prod. No. C-4255.) F. 100 mm Magnesium Chloride Solution (MgCl 2 ) (Prepare 5 ml in deionized water using Magnesium Chloride, Hexahydrate, Sigma Prod. No. M-0250.) G. PK/LDH Mixed Enzymes 1 (PK/LDH) (Use PK/LDH Enzymes Suspension, Sigma Stock No ) H. 100 mm Glycylglycine Buffer with 200 mm Potassium Chloride, ph 8.0 at 25 C (CPK Enz Diluent) (Prepare 25 ml in deionized water using Gly-Gly, Free Base, Sigma Prod. No. G-1002 and Potassium Chloride, Sigma Prod. No. P Adjust to ph 8.0 at 25 C with 1 M NaOH.) I. Creatine Phosphokinase Enzyme Solution (CPK) (Immediately before use, prepare a solution containing 900 units/ml of Creatine Phosphokinase, Sigma Prod. No. C-3755 in Reagent H.) J. Creatininase Enzyme Solution (Creatininase) (Immediately before use, prepare a solution containing unit/ml of Creatininase in cold Reagent A.) PROCEDURE: Pipette (in milliliters) the following reagents into suitable cuvettes: Test Blank SPCREA01 Page 2 of 4
3 Reagent A (Buffer) Reagent B (ß-NADH) Reagent C (ATP) SPCREA01 Page 3 of 4
4 PROCEDURE: (continued) Test Blank Reagent D (PEP) Reagent E (Substrate) Reagent F (MgCl 2 ) Reagent G (PK/LDH) Reagent I (CPK) Mix by inversion and equilibrate to 25 C. Monitor the A 340nm until constant, using a suitably thermostatted spectrophotometer. Then add: Reagent J (Creatininase) Reagent A (Buffer) Immediately mix by inversion and record the decrease in the A 340nm for approximately 5 minutes. Obtain the ra 340nm /minute using the maximum linear rate for both the Test and Blank. CALCULATION: Units/ml enzyme = (ra 340nm /min Test - ra 340nm /min Blank)(3.19)(df) (6.22)(0.05) 3.19 = Total Volume (in milliliters) of assay df = Dilution factor 6.22 = Millimolar extinction coefficient of ß-NADH at 340 nm 0.05 = Volume (in milliliter) of enzyme used units/ml enzyme Units/mg solid = mg solid/ml enzyme units/ml enzyme Units/mg protein = mg protein/ml enzyme UNIT DEFINITION: One unit will hydrolyze 1.0 µmole of creatinine to creatine per minute at ph 8.0 at 25 C. SPCREA01 Page 4 of 4
5 FINAL ASSAY CONCENTRATION: In a 3.19 ml reaction mix, the final concentrations are 88 mm glycylglycine, 0.38 mm ß-nicotinamide adenine dinucleotide, 1.0 mm adenosine 5'-triphosphate, 0.63 mm phospho(enol)pyruvate, 28 mm creatinine, 3.1 mm magnesium chloride, 28 units pyruvate kinase, 40 units lactic dehydrogenase, 90 units creatine phosphokinase, 6.1 mm potassium chloride, and unit creatininase. REFERENCE: NOTES: Bergmeyer, H.U., Gawehn, K. and Grassl, M.(1974) in Methods of Enzymatic Analysis (Bergmeyer, H.U. ed.) Vol. I, 2nd ed., , Academic Press, Inc., New York, NY 1. Contains not less than 700 Pyruvate Kinase units and 1000 Lactic Dehydrogenase units per ml. 2. Pyruvate Kinase Unit Definition: One unit will convert 1.0 µmole of phospho(enol)pyruvate to pyruvate per minute at ph 7.6 at 37 C. 3. L-Lactic Dehydrogenase Unit Definition: One unit will reduce 1.0 µmole of pyruvate to L-lactate per minute at ph 7.5 at 37 C. 4. Creatine Phosphokinase Unit Definition: One unit will transfer 1.0 µmole of phosphate from phosphocreatine to ADP per minute at ph 7.4 at 30 C. 5. This assay is based on the cited reference. 6. Where Sigma Product or Stock numbers are specified, equivalent reagents may be substituted. This procedure is for informational purposes. For a current copy of Sigma s quality control procedure contact our Technical Service Department. SPCREA01 Page 5 of 4
Enzymatic Assay of CHOLINE KINASE (EC )
Enzymatic Assay of CHOLINE KINASE PRINCIPLE: Choline + ATP CK > o-phosphocholine + ADP ADP + PEP PK > ATP + Pyruvate Pyruvate + ß-NADH LDH > Lactate + ß-NAD Abbreviations used: ATP = Adenosine 5'-Triphosphate
More informationEnzymatic Assay of PYRUVATE KINASE (EC ) From Rabbit Liver
Enzymatic Assay of PYRUVATE KINASE PRINCIPLE: Phospho(enol)pyruvate + ADP Pyruvate Kinase > Pyruvate + ATP Mg2 + Pyruvate + ß-NADH Lactic Dehydrogenase > Lactate + ß-NAD Abbreviations used: ADP = Adenosine
More informationEnzymatic Assay of GUANYLATE KINASE (EC )
PRINCIPLE: GMP + ATP Guanylate Kinase > GDP + ADP ADP + PEP Pyruvate Kinase > ATP + Pyruvate GDP + PEP Pyruvate Kinase > GTP + Pyruvate 2 Pyruvate + 2 ß-NADH Lactic Dehydrogenase > 2 Lactate + 2 ß-NAD
More informationEnzymatic Assay of GALACTOSYLTRANSFERASE (EC )
Enzymatic Assay of GALACTOSYLTRANSFERASE PRINCIPLE: UDP-Galactose + D-Glucose Galactosyltransferase > UDP + Lactose UDP + PEP PK > Pyruvate + UTP Pyruvate + ß-NADH LDH > Lactate + ß-NAD Abbreviations used:
More informationEnzymatic Assay of GLUCONATE KINASE (EC ) ß-NADPH = ß-Nicotinamide Adenine Dinucleotide Phosphate,
Enzymatic Assay of GLUCONATE KINASE PRINCIPLE: D-Gluconate + ATP Gluconate Kinase > 6-Phospho-D-Gluconate + ADP 6-Phospho-D-Gluconate + ß-NADP G-PGDH > D-Ribulose-5'-P + ß-NADPH + CO 2 Mg2+ Abbreviations
More informationEnzymatic Assay of PHOSPHODIESTERASE, 3':5'-CYCLIC NUCLEOTIDE Crude Complex
PRINCIPLE: 3':5'-cAMP + H 2 O PDE-3':5'-CN > AMP AMP + ATP Myokinase > 2 ADP 2 ADP + 2 PEP Pyruvate Kinase > 2 ATP + 2 Pyruvate 2 Pyruvate + 2 ß-NADH Lactic Dehydrogenase > 2 Lactate + 2 ß-NAD Abbreviations
More informationEnzymatic Assay of RIBOKINASE (EC )
PRINCIPLE: ATP + D-Ribose Ribokinase > ADP + D-Ribose 5-P D-Ribose 5-P PRI > D-Ribulose 5-P D-Ribulose 5-P Ru-5-P-3-Epim > D-Xylulose 5-P D-Xylulose 5-P + D-Ribose 5-P TK Mg++, Cocarboxylase > GAP + Sedoheptulose
More informationEnzymatic Assay of NAD-PYROPHOSPHORYLASE (EC )
Enzymatic Assay of NAD-PYROPHOSPHORYLASE PRINCIPLE: ß-NMN + ATP NAD-Pyrophosphorylase > ß-NAD + PP ß-NAD + Ethanol ADH > ß-NADH + Acetaldehyde Abbreviations used: ATP = Adenosine 5'-Triphosphate ADH =
More informationEnzymatic Assay of URIDINE 5'-DIPHOSPHOGALACTOSE 4-EPIMERASE (EC )
PRINCIPLE: UDP-Gal UDP-Gal 4-Epimerase > UDPG UDPG + 2 ß-NAD + H 2 O UDPG Dehydrogenase > UDP-Glucuronate + 2 ß-NADH Abbreviations used: UDP-Gal = Uridine 5'-Diphosphogalactose UDPG = Uridine 5'-Diphosphoglucose
More informationRUBISCO > 2 moles of 3-phosphoglycerate Mg +2
PRINCIPLE: RuDP + CO 2 RUBISCO > 2 moles of 3-phosphoglycerate Mg +2 3-Phosphoglycerate + ATP PGK > Glycerate 1,3-Diphosphate + ADP Glycerate 1,3-Diphosphate + ß-NADH GAPDH > Glyceraldehyde 3-Phosphate
More informationEnzymatic Assay of FRUCTOSE-6-PHOSPHATE KINASE, PYROPHOSPHATE DEPENDENT (EC ) from Mung Bean
PRINCIPLE: PP i + F-6-P PP i -PFK > F-1,6-DP + P i F-2,6-DP 1 F-1,6-DP Aldolase > GAP + DHAP GAP TPI > DHAP 2DHAP + 2 ß-NADH GDH > 2 Glycerol-3-Phosphate + 2 ß-NAD Abbreviations used: PP i = Pyrophosphate
More informationEnzymatic Assay of PHOSPHORYLASE KINASE (EC )
PRINCIPLE: Enzymatic Assay of PHOSPHORYLASE KINASE 2 Phosphorylase b + 4 ATP Phosphorylase Kinase > Phosphorylase a + 4 ADP Glycogen n + P i Phosphorylase a > Glycogen n-1 + a-d-glucose 1-Phosphate a-d-glucose
More informationEnzymatic Assay of PYRUVATE OXIDASE (EC ) from Pediococcus species
Enzymatic Assay of PYRUVATE OXIDASE PRINCIPLE: Pyruvate + O 2 + P i Pyruvate Oxidase > Acetylphosphate + CO 2 + H 2 O 2 FAD, TPP, Mg 2+ 2H2O2 + 4-Aminoantipyrine + N,N-Dimethylaniline POD > Quinonediimine
More informationEnzymatic Assay of ß-GALACTOSIDASE (EC ) from E. coli ß-Lactose as Substrate
PRINCIPLE: ß-Lactose + H 2 O ß-Galactosidase > D-Glucose + D-Galactose D-Glucose + O 2 + H 2 O Glucose Oxidase > D-Gluconic Acid + H 2 O 2 H 2 O 2 + o-dianisidine(reduced) Peroxidase > H 2 O + o-dianisidine(oxidized)
More informationEnzymatic Assay of RIBONUCLEIC ACID POLYMERASE 1 (EC )
PRINCIPLE: Enzymatic Assay of RIBONUCLEIC ACID POLYMERASE 1 DNA + NTP RNA Polymerase > DNA + RNA + PP i PP i + UDPG UDPG Pyrophosphorylase > UTP + Glucose 1-Phosphate Glucose 1-Phosphate Phosphoglucomutase
More informationEnzymatic Assay of CHOLESTEROL OXIDASE (EC )
PRINCIPLE: Cholesterol + O 2 Cholesterol Oxidase > H 2 O 2 + 4-Cholesten-3-One 2H 2 O 2 + 4-AAP + Phenol Peroxidase > 4 H 2 O + Quinoneimine Dye Abbreviation: 4-AAP = 4-Aminoantipyrine CONDITIONS: T =
More informationEnzymatic Assay of PHOSPHOLIPASE C (EC )
PRINCIPLE: Lecithin + H 2 O Phospholipase C > Diglyceride + Choline Phosphate Choline Phosphate + H 2 O Alkaline Phosphatase > Choline + P i Choline + O 2 Choline Oxidase > Betaine Aldehyde + H 2 O 2 Betaine
More informationB. 15 mm Ouabain Solution (Ouabain) (Prepare 10 ml in Reagent A using Ouabain Octahydrate, Sigma Prod. No. O3125.)
SIGMA QUALITY CONTROL TEST PROCEDURE Sigma Prod. No. A7510 PRINCIPLE: ATP + H 2 O ATPase > ADP + P i Abbreviations used: ATPase = Adenosine 5'-Triphosphatase ATP = Adenosine 5'-Triphosphate ADP = Adenosine
More informationB. 100 mm L-Glutamate Solution (L-Glu) (Prepare 2 ml in deionized water using L-Glutamic Acid, Monosodium Salt, Sigma Prod. No. G-1626.
Enzymatic Assay of L-GLUTAMATE OXIDASE PRINCIPLE: L-Glutamate + O 2 + H 2 O L-Glutamate Oxidase > a-ketoglutaric Acid + NH 3 + H 2 O 2 2 H 2 O 2 + H 2 O Catalase > 2 H 2 O + O 2 CONDITIONS: T = 30 C, ph
More informationEnzymatic Assay of PHOSPHOLIPASE C, PHOSPHATIDYLINOSITOL-SPECIFIC (EC )
PRINCIPLE: Step 1: Enzymatic Assay of PHOSPHOLIPASE C, PHOSPHATIDYLINOSITOL-SPECIFIC Acetylcholinesterase (membrane stroma bound) PLP C > Acetylcholinesterase (unbound) Step 2: ATI + DTNB Acetylcholinesterase
More informationEnzymatic Assay of PROTEASE (EC )
Enzymatic Assay of PROTEASE PRINCIPLE: Hemoglobin + H 2 O Protease > Amino Acids CONDITIONS: T = 37 C, ph = 2.8, A 660nm, Light path = 1 cm METHOD: Colorimetric REAGENTS: A. 50 mm Potassium Phthalate Buffer,
More informationEnzymatic Assay of PHOSPHOLIPASE C (EC ) from Bacillus cereus
PRINCIPLE: Lecithin + H 2 O Phospholipase C > Diglyceride + Choline Phosphate Choline Phosphate + H 2 O Alkaline Phosphatase > Choline + P i Choline + O 2 Choline Oxidase > Betaine Aldehyde + H 2 O 2 Betaine
More informationEnzymatic Assay of POLYGALACTURONASE (EC )
PRINCIPLE: Polygalacturonic Acid + H 2 O PG > Reducing Sugars Abbreviations: PG = Polygalacturonase CONDITIONS: T = 30 C, ph 5.0, A 540nm, Light path = 1 cm METHOD: Colorimetric REAGENTS: A. 50 mm Sodium
More informationB. 50 mm Calcium Chloride Solution (CaCl 2 ) (Prepare 25 ml in Reagent A using Calcium Chloride, Dihydrate, Sigma Prod. No. C-3881.
SIGMA QUALITY CONTROL TEST PROCEDURE ProductInformation Enzymatic Assay of PHOSPHOLIPASE C PRINCIPLE: L-α-Lecithin + H 2 O Phospholipase C > 1,2-Diglyceride + Choline Phosphate Choline phosphate + H 2
More informationEnzymatic Assay of ß-GLUCOSIDASE (EC )
PRINCIPLE: ß-D-Glucoside + H 2 O ß-Glucosidase > D-Glucose + an Alcohol CONDITIONS: T = 37 C, ph = 5.0, A 540nm, Light path = 1 cm METHOD: Colorimetric 1 REAGENTS: A. 100 mm Sodium Acetate Buffer, ph 5.0
More informationB. 1% (w/v) Salicin Substrate Solution (Salicin) (Prepare 50 ml in Reagent A using Salicin, Sigma Prod. No. S-0625.)
SIGMA QUALITY CONTROL TEST PROCEDURE (Q]\PDWLFÃ$VVD\ÃRIÃ */8&26,'$6( PRINCIPLE: 'Glucoside + H 2 O Glucosidase > D-Glucose + an Alcohol CONDITIONS: T = 37 C, ph = 5.0, A 540nm, Light path = 1 cm METHOD:
More information1.2 Systematic Name: Orthophosphoric-monoester phosphohydrolase (alkaline optimum)
Document Title Alkaline Phosphatase Page 1 of 6 Originating Department QA Approval Departments QA, QC Approval Date 5 th June 2017 Effective Date 8 th June 2017 1.0 PRODUCT DETAILS 1.1 Enzyme Name: Alkaline
More informationMethods of Enzyme Assay. By: Amal Alamri
Methods of Enzyme Assay By: Amal Alamri Introduction: All enzyme assays measure either the consumption of substrate or production of product over time. Different enzymes require different estimation methods
More informationTECHNICAL BULLETIN. Sialic Acid Quantitation Kit. Catalog Number SIALICQ Storage Temperature 2 8 C
Sialic Acid Quantitation Kit Catalog Number SIALICQ Storage Temperature 2 8 C TECHNICAL BULLETIN Product Description The Sialic Acid Quantitation Kit provides a rapid and accurate determination of total
More informationCELLULAR RESPIRATION SUMMARY EQUATION. C 6 H 12 O 6 + O 2 6CO2 + 6H 2 O + energy (ATP) STEPWISE REDOX REACTION
CELLULAR RESPIRATION SUMMARY EQUATION C 6 H 12 O 6 + O 2 6CO2 + 6H 2 O + energy (ATP) STEPWISE REDOX REACTION Oxidation: partial or complete loss of electrons Reduction: partial or complete gain of electrons
More informationNotes CELLULAR RESPIRATION SUMMARY EQUATION C 6 H 12 O 6 + O 2. 6CO 2 + 6H 2 O + energy (ATP) STEPWISE REDOX REACTION
AP BIOLOGY CELLULAR ENERGETICS ACTIVITY #2 Notes NAME DATE HOUR SUMMARY EQUATION CELLULAR RESPIRATION C 6 H 12 O 6 + O 2 6CO 2 + 6H 2 O + energy (ATP) STEPWISE REDOX REACTION Oxidation: partial or complete
More informationGLYCEROL
www.megazyme.com GLYCEROL ASSAY PROCEDURE K-GCROL 04/17 (*70 Assays per Kit) or (700 Microplate Assays per Kit) * The number of tests per kit can be doubled if all volumes are halved Megazyme 2017 INTRODUCTION:
More informationNotes CELLULAR RESPIRATION SUMMARY EQUATION C 6 H 12 O 6 + O 2. 6CO 2 + 6H 2 O + energy (ATP) STEPWISE REDOX REACTION
AP BIOLOGY CELLULAR ENERGETICS ACTIVITY #2 Notes NAME DATE HOUR SUMMARY EQUATION CELLULAR RESPIRATION C 6 H 12 O 6 + O 2 6CO 2 + 6H 2 O + energy (ATP) STEPWISE REDOX REACTION Oxidation: partial or complete
More informationenzymatic determinations of kecap Page 1 of 13
enzymatic determinations of kecap Page 1 of 13 Contains enzymatic determinations of: - citrate - glycerol - glucose, fructose, starch - galactose - L-glutamic acid - formic acid - malate - ethanol - acetate
More informationMethods of Enzyme Assay
Methods of Enzyme Assay Introduction All enzyme assays measure either the consumption of substrate or production of product over time. Different enzymes require different estimation methods dependingon
More informationKE-SIALIQ Sialic Acid Quantitation Kit. SialiQuant Sialic Acid Quantitation Kit
SialiQuant Sialic Acid Quantitation Kit Part Number KE-SIALIQ Certification of Analysis Lot Number 706.1A Kit Storage Kits should be stored at 4 C. Kit Contents Kit contains all the reagents to quickly
More informationWhat is respiration:
Respiration What is respiration: Aerobic respiration is the controlled release of energy from food using oxygen. The food involved in respiration is glucose. The energy is trapped in molecules of ATP.
More informationYeast Extracts containing Mannoproteins (Tentative)
0 out of 6 Residue Monograph prepared by the meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 84th meeting 2017 Yeast Extracts containing Mannoproteins (Tentative) This monograph
More informationMALTOSE, SUCROSE and D-GLUCOSE
www.megazyme.com MALTOSE, SUCROSE and D-GLUCOSE ASSAY PROCEDURE K-MASUG 11/16 (*34 Assays of each per Kit) * The number of tests per kit can be doubled if all volumes are halved Megazyme 2016 INTRODUCTION:
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 informationMaterials and Reagents
Extraction and Measurement the Activities of Cytosolic Phosphoenolpyruvate Carboxykinase (PEPCK) and Plastidic NADP-dependent Malic Enzyme (ME) on Tomato (Solanum lycopersicum) Sonia Osorio 1*, José G.
More informationCoupled, interconnecting reactions
Metabolism: Basic concepts Hand-out for the CBT version November 2011 This module is based on 'Biochemistry' by Berg, Tymoczko and Stryer, seventh edition (2011), Chapter 15: Metabolism: Basic Concepts
More informationUnit 2: Metabolic Processes
How is energy obtained biologically? Recall: Red Ox Reactions Unit 2: Metabolic Processes Oxidation Is the chief mechanism by which chemical potential energy is released This energy comes from reduced
More informationReview of Carbohydrate Digestion
Review of Carbohydrate Digestion Glycolysis Glycolysis is a nine step biochemical pathway that oxidizes glucose into two molecules of pyruvic acid. During this process, energy is released and some of it
More informationSUCROSE, D-FRUCTOSE and D-GLUCOSE
www.megazyme.com SUCROSE, D-FRUCTOSE and D-GLUCOSE ASSAY PROCEDURE K-SUFRG 04/17 (*100 Assays of each per Kit) * The number of tests per kit can be doubled if all volumes are halved Megazyme 2017 INTRODUCTION:
More informationGlycolysis by Human Spermatozoa: Levels of Glycolytic Intermediates
BIOLOGY OF REPRODUCrION 5, 221-227 (1971) Glycolysis by Human Spermatozoa: Levels of Glycolytic Intermediates R. N. PETERSON AND M. FREUND Laboratory of Reproductive Pharmacology, Departments of Pharmacology
More informationPhosFree TM Phosphate Assay Biochem Kit
PhosFree TM Phosphate Assay Biochem Kit (Cat. # BK050) ORDERING INFORMATION To order by phone: (303) - 322-2254 To order by Fax: (303) - 322-2257 To order by e-mail: cservice@cytoskeleton.com Technical
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 informationDate... Name... Group... Urine sample (Tube No 2)
Date... Name... Group... Instructions for the practical lesson on biochemistry Topic: Non-protein nitrogen compounds Task 1: Estimation of creatinine in serum and urine 1. Trichloroacetic acid 1.22 mol/l
More informationBioenergetics. Chapter 3. Objectives. Objectives. Introduction. Photosynthesis. Energy Forms
Objectives Chapter 3 Bioenergetics Discuss the function of cell membrane, nucleus, & mitochondria Define: endergonic, exergonic, coupled reactions & bioenergetics Describe how enzymes work Discuss nutrients
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 informationChapter 13 Carbohydrate Metabolism
Chapter 13 Carbohydrate Metabolism Metabolism of Foods Food is broken down into carbohydrates, lipids, and proteins and sent through catabolic pathways to produce energy. Glycolysis glucose 2 P i 2 ADP
More informationDeficiencies of Glycolytic Pathway
Deficiencies of Glycolytic Pathway -Mature RBCs have the capacity for a limited number of enzymatic reactions -The mature RBC is completely dependent on glucose as a source of energy. Glucose usually (90%)
More informationExperiment 9. NATURE OF α-amylase ACTIVITY ON STARCH
Experiment 9 NATURE OF α-amylase ACTIVITY ON STARC In Experiment 1 we described the action of α-amylase on starch as that of catalyzing the hydrolysis of α-1,4-glucosidic bonds at random in the interior
More informationCreatinine (serum, plasma)
Creatinine (serum, plasma) 1 Name and description of analyte 1.1 Name of analyte Creatinine 1.2 Alternative names None 1.3 Description of analyte Creatinine is a heterocyclic nitrogenous compound (IUPAC
More informationBiology 30 Structure & Function of Cells (Part 2) Bioenergetics: Energy: Potential energy: Examples: Kinetic energy. Examples:
Biology 30 Structure & Function of Cells (Part 2) Bioenergetics: Energy: Potential energy: Examples: Kinetic energy Examples: Energy can be transformed: Thermodynamics: First law of Thermodynamics: Second
More informationIntroduction to Metabolism Cell Structure and Function
Introduction to Metabolism Cell Structure and Function Cells can be divided into two primary types prokaryotes - Almost all prokaryotes are bacteria eukaryotes - Eukaryotes include all cells of multicellular
More information3.7.1 Define cell respiration [Cell respiration is the controlled release of energy from organic compounds in cells to form ATP]
3.7 Cell respiration ( Chapter 9 in Campbell's book) 3.7.1 Define cell respiration [Cell respiration is the controlled release of energy from organic compounds in cells to form ATP] Organic compounds store
More informationΒ-FRUCTOFURANOSIDASE ENZYME
KINETICS ANALYSIS OF Β-FRUCTOFURANOSIDASE ENZYME 2-The effects of enzyme concentration on the rate of an enzyme catalyzed reaction. Systematic names and numbers β-fructofuranosidase (EC 3.2.1.26) Reactions
More informationGPx Equation 1 Æ R - O - H + GSSG + H2 O
OXFORD BIOMEDICAL RESEARCH P.O. Box 522, Oxford MI 48371 USA USA: 800-692-4633 Fax: 248-852-4466 www.oxfordbiomed.com Colorimetric Assay for Cellular Glutathione Peroxidase Product No. FR 17 For Research
More informationNWLSS TM. Assay. Glutathione Peroxidase. Product NWK-GPX01 For Research Use Only
Premier Products for Superior Life Science Research NWLSS TM Glutathione Peroxidase Assay Product NWK-GPX01 For Research Use Only Simple assay kit for quantitative measurement of glutathione peroxidase
More informationUser s Manual and Instructions
User s Manual and Instructions Mitochondria Activity Assay (Cytochrome C Oxidase Activity Assay) Kit Catalog Number: KC310100 Introduction Mitochondria are the eukaryotic subcellular organelles that contain
More informationMitochondria and ATP Synthesis
Mitochondria and ATP Synthesis Mitochondria and ATP Synthesis 1. Mitochondria are sites of ATP synthesis in cells. 2. ATP is used to do work; i.e. ATP is an energy source. 3. ATP hydrolysis releases energy
More informationLecture'22:'April'30,'2013 Ch.%29:%Metabolism,%catabolism,%anabolism Metabolic%energy%&%ATP%Coupling Glycolysis%and%the%Link%ReacDon
CM'224' 'rganic'chemistry'ii pring'2013,'des'laines' 'rof.'chad'landrie 2 2 2 Gº' = -7.3 + + + Lecture'22:'April'30,'2013 Ch.%29:%Metabolism,%catabolism,%anabolism Metabolic%energy%&%AT%Coupling Glycolysis%and%the%Link%ReacDon
More informationWhole muscle assays. agpdh ADP --ATP. PEP Pyruvate +Lactate NADH N~D+
A novel method for measurement of triglyceride lipase activity: suitable for microgram and nanogram quantities of tissue Douglas A. Young, Douglas S. King, May Chen, Beverly Norris,' and Patti M. Nemeth*
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 informationRegulation of glycolysis/fructolysis in buffalo
Regulation of glycolysis/fructolysis in buffalo spermatozoa K. K. Gandhi and S. R. Anand National Dairy Research Institute, Karnal-132001, India Summary. Assay of maximal activities of 11 glycolytic enzymes
More informationIntroduction to Carbohydrate metabolism
Introduction to Carbohydrate metabolism Some metabolic pathways of carbohydrates 1- Glycolysis 2- Krebs cycle 3- Glycogenesis 4- Glycogenolysis 5- Glyconeogenesis - Pentose Phosphate Pathway (PPP) - Curi
More informationCellular Respiration. Cellular Respiration. C 6 H 12 O 6 + 6O > 6CO 2 + 6H energy. Heat + ATP. You need to know this!
Cellular Respiration LISA Biology Cellular Respiration C 6 H 12 O 6 + 6O 2 - - - - - > 6CO 2 + 6H 2 0 + energy You need to know this! Heat + ATP 1 Did that equation look familiar? * The equation for cellular
More informationD-FRUCTOSE and D-GLUCOSE
www.megazyme.com DFRUCTOSE and DGLUCOSE (Liquid Stable Format) ASSAY PROCEDURE KFRGLQR 02/17 (1100 AutoAnalyser Assays of each per Kit) or (1100 Microplate Assays of each per Kit) Megazyme 2017 INTRODUCTION:
More informationData sheet. TBARS Assay kit. (Colorimetric/Fluorometric) Kit Contents. MDA-TBA Adduct. 2-Thiobarbituric Acid. Cat. No: CA995.
Data sheet Cat. No: CA995 TBARS Assay kit (Colorimetric/Fluorometric) Introduction Oxidative stress in the cellular environment results in the formation of highly reactive and unstable lipid hydroperoxides.
More informationAdenosine triphosphate (ATP)
Adenosine triphosphate (ATP) 1 High energy bonds ATP adenosine triphosphate N NH 2 N -O O P O O P O- O- O O P O- O CH 2 H O H N N adenine phosphoanhydride bonds (~) H OH ribose H OH Phosphoanhydride bonds
More informationBioenergetics: Energy for Exercise. Chapter 3 pp 28-47
Bioenergetics: Energy for Exercise Chapter 3 pp 28-47 ATP ATP Adenosine Ribose 3 Phosphates ATP ATP ATP ATP ATP ATP ATP ATP ATP Energy Out ADP + P ATP ATP Energy Out ADP + P ATP ATP Energy Out Energy In
More informationCHE 242 Exam 3 Practice Questions
CHE 242 Exam 3 Practice Questions Glucose metabolism 1. Below is depicted glucose catabolism. Indicate on the pathways the following: A) which reaction(s) of glycolysis are irreversible B) where energy
More informationBASIC ENZYMOLOGY 1.1
BASIC ENZYMOLOGY 1.1 1.2 BASIC ENZYMOLOGY INTRODUCTION Enzymes are synthesized by all living organisms including man. These life essential substances accelerate the numerous metabolic reactions upon which
More informationGB Translated English of Chinese Standard: GB NATIONAL STANDARD OF THE
Translated English of Chinese Standard: GB5009.85-2016 www.chinesestandard.net Buy True-PDF Auto-delivery. Sales@ChineseStandard.net GB NATIONAL STANDARD OF THE PEOPLE S REPUBLIC OF CHINA GB 5009.85-2016
More informationTABLE OF CONTENTS GENERAL INFORMATION... 1
BIOO RESEARCH PRODUCTS Glucose Assay Kit Manual Catalog #: 5611-01 BIOO Scientific Corp. 2011 TABLE OF CONTENTS GENERAL INFORMATION... 1 Product Description... 1 Procedure Overview... 1 Required Materials
More informationChap 3 Metabolism and Growth
Chap 3 Metabolism and Growth I. Metabolism Definitions: Metabolism includes two parts: anabolism and catabolism Catabolism: Anabolism: Aerobic metabolism: catabolism anabolis m catabolis anabolis m Anaerobic
More informationHigher Biology. Unit 2: Metabolism and Survival Topic 2: Respiration. Page 1 of 25
Higher Biology Unit 2: Metabolism and Survival Topic 2: Respiration Page 1 of 25 Sub Topic: Respiration I can state that: All living cells carry out respiration. ATP is the energy currency of the cell
More informationCHAPTER 24: Carbohydrate, Lipid, & Protein Metabolism. General, Organic, & Biological Chemistry Janice Gorzynski Smith
CHAPTER 24: Carbohydrate, Lipid, & Protein Metabolism General, Organic, & Biological Chemistry Janice Gorzynski Smith CHAPTER 24: Carbohydrate, Lipid, & Protein Metabolism Learning Objectives: q Role in
More informationMIXED XYLANASE, β-glucanase ENZYME PREPARATION, produced by a strain of HUMICOLA INSOLENS
MIXED XYLANASE, β-glucanase ENZYME PREPARATION, produced by a strain of HUMICOLA INSOLENS New specifications prepared at the 61st JECFA (2003) and published in FNP 52 Add 11 (2003). An ADI not specified
More informationCHEM121 Unit 2: Carbohydrate Metabolism
CHEM121 Unit 2: Carbohydrate Metabolism Lecture 3 At the end of the lecture, students should be able to: Define metabolism Discuss the structure and function of ATP in metabolism Discuss glycolysis in
More informationChapter 5 MITOCHONDRIA AND RESPIRATION 5-1
Chapter 5 MITOCHONDRIA AND RESPIRATION All organisms must transform energy. This energy is required to maintain a dynamic steady state, homeostasis, and to insure continued survival. As will be discussed
More informationRENAL FUNCTION ASSESSMENT ASSESSMENT OF GLOMERULAR FUNCTION ASSESSMENT OF TUBULAR FUNCTION
Measured GFR (mgfr mgfr) and Estimated t GFR (egfr egfr) R. Mohammadi Biochemist (Ph.D.) Faculty member of Medical Faculty RENAL FUNCTION ASSESSMENT ASSESSMENT OF GLOMERULAR FUNCTION ASSESSMENT OF TUBULAR
More informationAn organic catalysts that enhances the speed or likelihood of a bio-chemical reaction by lowering the energy of activation.
Micro 260 Fall 2009 Name: Allan Keys Tools: You may use your notes and or book for this assignment 1) What is an enzyme? (4 pts) An organic catalysts that enhances the speed or likelihood of a bio-chemical
More informationA. Incorrect! No, this statement is accurate so is not the correct selection to the question.
Biochemistry - Problem Drill 14: Glycolysis No. 1 of 10 1. Which of the following statements is incorrect with respect to glycolysis? (A) It is the conversion of glucose to pyruvate. (B) In glycolysis
More informationab HRP Substrate Instructions for Use
ab155473 HRP Substrate Instructions for Use For detecting the activity of many oxidases, related enzymes/substrates or cofactors such as glucose, acetylcholine and cholesterol, L-glutamate, amino acids
More information4.2 Glycolysis. ATP in Living Systems. ATP Structure and Function. Page by: OpenStax. Summary
4.2 Glycolysis Page by: OpenStax Summary Even exergonic, energy-releasing reactions require a small amount of activation energy to proceed. However, consider endergonic reactions, which require much more
More informationExperiment 3: Activity Determination
Experiment 3: Activity Determination Introduction: Specific activity is a method for measuring enzymatic activity and the enzyme purity in a mixture. In order to determine the specific activity of an enzyme,
More informationBCH 4053 THIRD EXAM November 5, 1999
BCH 4053 THIRD EXAM November 5, 1999 I remind you that you are bound by the Academic Honor Code. This means (1) you will not give or receive information during this exam, nor will you consult with unauthorized
More informationFluorometric procedures for measuring triglyceride concentrations in small amounts of tissue and plasma
Fluorometric procedures for measuring triglyceride concentrations in small amounts of tissue and plasma P. M. Nemeth,' 0. E. Hitchins,' L. Solanki, and T. G. Cole* Departments of Neumlogy, Anatomy, and
More informationTECHNICAL INFORMATION
MP Biomedicals, LLC 29525 Fountain Parkway Solon, Ohio 44139 TECHNICAL INFORMATION Telephone: 440/337-1200 Toll Free: 800/854-0530 Fax: 440/337-1180 mailto: biotech@mpbio.com web: http://www.mpbio.com
More informationMicro Bioactivity Analyzer
Micro Bioactivity Analyzer AMIS-11 Application Data Bio-X Inc. October 28 Micro Bioactivity Analyzer MIS-11 enables the real-time base quantitative measurement of bioactive reactions such as enzyme reaction,
More informationNWLSS TM Glutathione Reductase Activity (Microplate Assay)
Premier Products for Superior Life Science Research NWLSS TM Glutathione Reductase Activity (Microplate Assay) Product NWK-GR01 For Research Use Only Simple assay kit for measurement of Glutathione Reductase
More informationMetabolic Energy Part 2 الفريق الطبي األكاديمي
Metabolic Energy Part 2 الفريق الطبي األكاديمي Done By :- Shady Soghayr Corrected By :- Rasha Rakan لكية الطب البرشي البلقاء التطبيقية / املركز 2022 2016/ Metabolic Energy Page 1 Metabolic energy For aa
More informationChapter 27 Bioenergetics; How the Body Converts Food to Energy
Chapter 27 Bioenergetics; How the Body Converts Food to Energy 1 Metabolism Metabolism: The sum of all chemical reactions involved in maintaining the dynamic state of a cell or organism. Pathway: A series
More informationDIFFERENCES BETWEEN ALI & BEGM
Table of Contents DIFFERENCES BETWEEN ALI & BEGM... 1 MEDIA PREPARATION (ALI & BEGM)... 2 ADDITIVES FOR BEGM AND ALI... 2 STOCK 11... 3 STOCK 4... 3 ETHANOLAMINE 3 BOVINE PITUITARY EXTRACT... 3 HYDROCORTISONE...
More informationBIOO LIFE SCIENCE PRODUCTS
BIOO LIFE SCIENCE PRODUCTS FOR REFERENCE PURPOSES This manual is for Reference Purposes Only. DO NOT use this protocol to run your assays. Periodically, optimizations and revisions are made to the kit
More information