Human Soluble TNFR II, sTNFR II ELISA Kit

Catalog No.: BEK1217

Size: 96T

Range: 7.8 pg/ml-500 pg/ml

Sensitivity < 2 pg/ml

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

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

Introduction

Tumor necrosis factor receptor 2 (TNFR2), also known as tumor necrosis factor receptor superfamily member 1B (TNFRSF1B) and CD120b, is a membrane receptor that binds tumor necrosis factor-alpha (TNFɑ). The protein encoded by this gene is a member of the tumor necrosis factor receptor superfamily, which also contains TNFRSF1A. Tumor necrosis factor receptor superfamily member 1B is a protein that in humans is encoded by the TNFRSF1B gene. This gene contains 10 exons and spans about 26 kb. It is present on many cell types, especially those of myeloid origin, and is strongly expressed on stimulated T and B lymphocytes. It is a member of the Tumor necrosis factor receptor superfamily, which also contains TNFRSF1A. It causes the ubiquitination of TRAF2 by CIAP1, which can play a proapoptotic role in TNF signaling.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-sTNFR II polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-sTNFR II 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 sTNFR II amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of sTNFR II can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human sTNFR II antibody
2. Lyophilized Human sTNFR II standards: 2 tubes (10ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human sTNFR II 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