Mouse B7-1/CD80 ELISA Kit

Catalog No.: BEK1031

Size: 96T

Range: 62.5 pg/ml-4000 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 B7-1 in tissue lysates or cell culture supernates. The significance of detection in serum, plasma, body

fluids is undetermined.

Introduction

Cluster of Differentiation 80 (also CD80 and B7-1) is a protein found on activated B cells and monocytes. B7-1 provides regulatory signals for T lymphocytes as a consequence of binding to the CD28 and CTLA4 ligands of T cells. The CD80 gene has 6 exons that span approximately 32 kb of genomic DNA. The antigen presentation coactivators B7-1 and B7-2 may be important for lymphocytic infiltration and the immune response against thyroid carcinoma.

Principle of the Assay  

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

Kit components

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