Kinex Reverse Lysate Microarray KRLM-1.0. Case Study

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Alicia Sarah Leonard
5 years ago
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1 Kinex Reverse Lysate Microarray KRLM-1. Case Study Probing with an Anti-phospho-ERK1/2 Antibody

2 The Kinex Reverse Lysate Microarray (KRLM) is a powerful tool for rapid characterization of a protein for which a highly specific antibody is available. This unique microarray features about 25 different lysates prepared from several human tumour cell lines that have been treated with diverse hormones and drugs in time course and dose response experiments as well as tissues from monkey, rat and mouse. Each lysate has been printed in triplicate at four different concentrations to ensure that detection of the antigen is within the linear range. For this presentation, we have provided the results from the probing of one KRLM-1. microarray with an phospho-site-specific antibody that targets the activations sites of the MAP kinases ERK1 and ERK2. These kinase are phosphorylated in response to diverse growth factors and during oocyte maturation. For comparison, we have provided some of the results from parallel immunoblotting studies with similar cell and tissue extracts. Any interesting results from the KRLM-1. analysis can be confirmed with our Kinetworks Custom Sample Screen KCSS 1. immunoblotting service. With the KRLM-1. service, the results are provided in a Microsoft Excel spreadsheet. For an additional US $3, Kinexus can prepare a series of Powerpoint Slides with the results from a single KRLM-1. analysis as exemplied here. Individual graphs can be prepared for US $2 each. For more information, contact our Technical Services representatives toll free in North America at KINEXUS or by at info@kinexus.ca.

3 Cell Line: A431 Human Epidermoid Carcinoma Treatment: Epidermal Growth Factor 9 Conc.: 1 ng/ml 2.5 min 5 min 1 min 2 min 2.5 mg/ml 1.25 mg/ml.625 mg/ml.3125 mg/ml 4 min 8 min

4 Cell Line: A431 Human Epidermoid Carcinoma Treatment: Epidermal Growth Factor Starved Control Time: 5 min 25 Dose Response mg/ml 1.6 mg/ml.8 mg/ml.4 mg/ml 4 8

5 Cell Line: MCF7 Human Breast Adenocarcinoma Treatment: Insulin Conc.: 1 µg/ml 2.5 min 5 min 1 min 2 min 4 min 3.2 mg/ml 1.6 mg/ml.8 mg/ml.4 mg/ml 8 min

6 Cell Line: MCF7 Human Breast Adenocarcinoma Treatment: Insulin 1 1 Time: 2.5 min 2.5 Dose Response mg/ml.4375 mg/ml 8

7 Cell Line: T98G Human Brain Glioblastoma Treatment: Platelet Derived Growth Factor Conc.: 5 ng/ml 2.5 min 5 min 1 min 2 min 2.9 mg/ml 1.45 mg/ml.725 mg/ml.3625 mg/ml 4 min 8 min

8 Cell Line: T98G Human Brain Glioblastoma Treatment: Platelet Derived Growth Factor 1 1 Time: 5 min 6.25 Dose Response mg/ml.4375 mg/ml 1 2

9 Cell Line: HepG2 Human Liver Carcinoma Treatment: Insulin Conc.: 1 µg/ml Time: 5 min 2.5 min min 1 min 2 min Dose Response min 8 min.875 mg/ml.4375 mg/ml mg/ml 1.7 mg/ml.85 mg/ml.425 mg/ml

10 Cell Line: A549 Human Lung Carcinoma Treatment: Interferon-gamma 9 Conc.: 5 ng/ml Time: 8 min 2 hr 25 4 hr 8 hr 16 hr Dose Response hr 48 hr 1.8 mg/ml.9 mg/ml.45 mg/ml.225 mg/ml mg/ml 1.1 mg/ml.55 mg/ml.275 mg/ml

11 Cell Line: HL6 Human Promyelocytic Leukemia Treatment: Staurosporine Conc. 1 µg/ml Time: 16 hr 2 hr.63 4 hr 8 hr 16 hr Dose Response hr 48 hr 2.5 mg/ml 1.25 mg/ml.675 mg/ml.3375 mg/ml mg/ml 1.6 mg/ml.8 mg/ml.4 mg/ml

12 Cell Line: HCT-116 Human Colon Carcinoma Treatment: Nocodazole 9 Conc.: 1 ng/ml Time: 16 h 2 hr 25 4 hr 8 hr 16 hr Dose Response hr 4 48 hr.875 mg/ml.4375 mg/ml mg/ml.4375 mg/ml

13 Cell Line: HEK-293 Human Embryonic Kidney Treatment: Anisomycin Conc.: 1 µg/ml 2.5 min 5 min 1 min 2 min 4 min 8 min 2.6 mg/ml 1.3 mg/ml.65 mg/ml.325 mg/ml

14 Rhesus Monkey Tissues - Untreated Bladder Male Fad Pads Heart Intestines Kidney Liver Skeletal muscle.875 mg/ml.4375 mg/ml Spinal cord Spleen Stomach Testes

15 Rat Tissues - Untreated Brain Male Brain Female Heart Heart Kidney Kidney Liver Liver Lung Lung Skeletal muscle Skeletal muscle Spleen Spleen Thymus Testes.875 mg/ml.4375 mg/ml Thymus Ovary.875 mg/ml.4375 mg/ml Male Rat Tissues

16 Mouse Tissues - Untreated Bladder Male Bladder Female Brain Brain Heart Heart Kidney Kidney Liver Liver Lung Lung Skeletal muscle Skeletal muscle Spleen Stomach Seminal vesicle Testes.875 mg/ml.4375 mg/ml Spleen Stomach Ovary Uterus.875 mg/ml.4375 mg/ml Female MouseTissues

17 Primary Cell: Xenopus laevis Frog Oocyte - Stage VI Treatment: Progesterone 1 Immature Immature Immature 4 hr 5 hr 6 hr 7 hr 8 hr 9 hr GVBD+ GVBD+ GVBD+ The results from the microarray analysis of 48 individual frog oocytes are shown at only one dilution in triplicate. Note the high degree of variation in Erk2 phosphorylation despite the fact that all of the oocytes were from the same frog. Oocyte 1 Oocyte 2 Oocyte 3 Oocyte 4

SOMAPLEX REVERSE PHASE PROTEIN MICROARRAY HUMAN KIDNEY TUMOR & NORMAL TISSUE

SOMAPLEX REVERSE PHASE PROTEIN MICROARRAY HUMAN KIDNEY TUMOR & NORMAL TISSUE 45 CLINICAL CASES SERIAL DILUTION MULTIPLE PROTEIN CONCENTRATION QUANTITATIVE ASSAY PRODUCT NUMBER: PM1-001-N SOMAPLEX REVERSE