Human B7-1 / CD80 ELISA Kit

Catalog No.: BEK1345

Size: 96T

Range: 62.5 pg/ml - 4000 pg/ml

Sensitivity < 12 pg/ml

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

Application: For quantitative detection of B7-1 / CD80 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

The Cluster of differentiation 80 (also CD80 and B7-1) is a B7, type I membrane protein in the immunoglobulin superfamily, with an extracellular immunoglobulin constant-like domain and a variable-like domain required for receptor binding. It is closely related to CD86, another B7 protein (B7-2), and often works in tandem. Both CD80 and CD86 interact with costimulatory receptors CD28, CTLA-4 (CD152) and the p75 neurotrophin receptor. CD80 is a member of the B7 family, which consists of molecules present at APCs and their receptors present on the T-cells. CD80 is present specifically on DC, activated B-cells, and macrophages, but also T-cells. CD80 is also a transmembrane glycoprotein and a member of the Ig superfamily. 

Principle of the Assay

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- B7-1 / CD80 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- B7-1 / CD80 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 B7-1 / CD80 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of B7-1 / CD80 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human B7-1 / CD80 antibody
2. Lyophilized human B7-1 / CD80 standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human B7-1 / CD80 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
11. 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