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Analysis of CD40/TNFRSF5 (Mouse) by ELISA (CAT#: STEM-MB-0727-LGZ)

Introduction

CD40 is a 48kda type I transmembrane protein, which is an important immune cell communication mediator connecting innate immunity and adaptive immunity. CD40 is present on platelets, B cells and myeloid cells, but also on non-hematopoietic cells such as endothelial cells, fibroblasts, smooth muscle cells and even some types of tumor cells. The cognate ligand of CD40 is CD154 (TNFSF5/CD40L), a 39 kDa type II transmembrane protein. CD40L expression is usually inducible and restricted to cells of the hematopoietic system, such as platelets, granulocytes, activated T cells, activated B cells, and activated natural killer (NK) cells, but is also weakly expressed by endothelial cells and smooth muscle cells.




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

Oncology

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: Serum, plasma or cell culture supernatant

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