Human Soluble TNFR I, sTNFR I ELISA Kit

Catalog No.: BEK1216

Size: 96T

Range: 7.8 pg/ml-500 pg/ml

Sensitivity < 1 pg/ml

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

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

Introduction

Tumor necrosis factor receptor 1 (TNFR1), also known as tumor necrosis factor receptor superfamily member 1A (TNFRSF1A) and CD120a, is a ubiquitous membrane receptor that binds tumor necrosis factor-alpha (TNFɑ). Tumor necrosis factor receptor superfamily member 1A is a protein that in humans is encoded by the TNFRSF1A gene, which chromosome 12p13.2. It is a member of the Tumor necrosis factor receptor superfamily, which also contains TNFRSF1B. Chan et al. (2000) found that, in contrast, the p60 and p80 TNFA receptors self-assemble through a distinct functional domain in the TNFR extracellular domain, termed the pre-ligand assembly domain (PLAD), in the absence of ligand.

Principle of the Assay  

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

Kit components

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