Human CD137 / TNFRSF9 ELISA Kit

Catalog No.: BEK1303

Size: 96T

Range: 31.2 pg/ml - 2000 pg/ml

Sensitivity < 10 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months. 

Application: For quantitative detection of CD137 / TNFRSF9 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

CD137, a member of the tumor necrosis factor (TNF) receptor family, is a type 1 transmembrane protein, expressed on surfaces of leukocytes and non-immune cells. Its alternative names are tumor necrosis factor receptor superfamily member 9 (TNFRSF9), 4-1BB, and induced by lymphocyte activation (ILA). It is of interest to immunologists as a co-stimulatory immune checkpoint molecule, and as a potential target in cancer immunotherapy. CD137 is only expressed on the cell surface after T cell activation. When T cells are activated by Antigen Presenting Cells (APCs), CD137 becomes embedded in CD4+ and CD8+ T cells.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- CD137 / TNFRSF9 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- CD137 / TNFRSF9 polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the CD137 / TNFRSF9 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of CD137 / TNFRSF9 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human CD137 / TNFRSF9 antibody
2. Lyophilized human CD137 / TNFRSF9 standards: 2 tubes (10ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human CD137 / TNFRSF9 antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L