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Analysis of STAT5a/b (Phosphoproteins , Intracellular Signaling) by ELISA (CAT#: STEM-MB-1190-LGZ)

Introduction

Signal transducer and activator of transcription 5 (STAT5) is a member of the Jak/STAT signaling pathway and is activated by various cytokines (IL22, IL6, IFN-α). STAT5 has two isoforms (A and B) that share 93% amino acid identity and bind the DNA consensus site TTCN3GAA. STAT5 acts as a signal transducer in the cytoplasm to mediate cytokine signaling, translocates to the nucleus after phosphorylation, and activates the transcription of specific genes. STAT5 is involved in a series of biological processes from regulation of apoptosis to proliferation, differentiation and survival of adult mammary gland.




Principle

Enzyme-linked immunosorbent assay (ELISA) is an enzyme-labeled solid phase immunoassay technique. Its basic principle is to bind the antigen (or antibody) to the solid phase carrier, and the antigen (or antibody) and a certain enzyme link to enzyme labeled antigen (or antibody). During detection, the sample to be tested and the enzymic antigen (or antibody) react with the antigen (or antibody) on the solid phase carrier according to certain procedures, and then remove the unreacted part by washing method. After adding the substrate, the substrate is catalyzed by the enzyme on the solid phase carrier to produce colored substances. Through qualitative or quantitative detection of the amount of colored products, the content of the substance to be measured in the sample can be determined.

Applications

Phosphoproteins, Intracellular Signaling

Procedure

1. Add standards or samples to each well and incubate.
2. Pour off the liquid in the well, biotinylated antibody working solution and incubate.
3. Add enzyme conjugate working solution and incubate.
4. Add substrate TMB and incubate.
5. Add stop solution and measure OD value.
6. Calculation of results.

Materials

• Sample Type: Cell culture extracts, Cell Lysate
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