B7-2/CD86 Antibody Pair [HRP]

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Sandwich ELISA: B7-2/CD86 Antibody Pair [HRP] [NBP2-79624] - This standard curve is only for demonstration purposes. A standard curve should be generated for each assay.

Product Details

Summary
Reactivity RtSpecies Glossary
Applications WB, ELISA, ICC/IF, S-ELISA
Clonality
Monoclonal
Host
Mouse
Conjugate
HRP
Standard Curve Range
4.69-300 pg/ml (example only; lot dependent)
Sensitivity
4.7 pg/ml (example only; lot dependent)

Order Details

B7-2/CD86 Antibody Pair [HRP] Summary

Description
Solid Phase sandwich ELISA for the quantitative determination of Rat B7-2/CD86.
Specificity
This solid phase sandwich ELISA utilizes a monoclonal antibody specific for Rat B7-2/CD86.
Standard Curve Range
4.69-300 pg/ml (example only; lot dependent)
Sensitivity
4.7 pg/ml (example only; lot dependent)
Assay Type
Sandwich ELISA
Sample Volume
100 ul
Clonality
Monoclonal
Host
Mouse
Gene
CD86

Applications/Dilutions

Dilutions
  • ELISA
  • Immunocytochemistry/Immunofluorescence
  • Sandwich ELISA
  • Western Blot
Application Notes
1. Capture Antibody: Dilute to a working concentration of 2 ug/ml in CBS before coating.
2. Detection Antibody: Dilute to working concentration of 0.5 ug/ml in detection antibody dilution buffer before use.
3. Standard: Reconstitute with 1 ml detection antibody dilution buffer. After reconstitution, store at -20C to -80C in a manual defrost freezer. A seven-point standard curve using 2-fold serial dilutions in sample dilution buffer, and a high standard of 0.3 ng/ml is recommended.

Packaging, Storage & Formulations

Storage
Storage of components varies. See protocol for specific instructions.

Kit Components

Components
  1. Capture Antibody: 0.4 mg/ml of mouse anti-rat B7-2/CD86 monoclonal antibody.
  2. Detection Antibody: 0.25 mg/ml mouse anti-B7-2/CD86 monoclonal antibody conjugated to horseradish-peroxidase (HRP).
  3. Standard: Each vial contains 19 ng of recombinant rat B7-2/CD86.

Alternate Names for B7-2/CD86 Antibody Pair [HRP]

  • Activation B7-2 antigen
  • B70
  • B7-2 antigen
  • B72
  • B7-2
  • B-lymphocyte activation antigen B7-2
  • BU63
  • CD28 antigen ligand 2
  • CD28LG2B7-2 antigen)
  • CD86 antigen
  • CD86 molecule
  • CD86
  • CTLA-4 counter-receptor B7.2
  • FUN-1
  • LAB72
  • MGC34413
  • T-lymphocyte activation antigen CD86

Background

B7-2, also referred to as CD86, is a glycosylated type 1 membrane protein that is a member of the immunoglobulin (Ig) superfamily (1). B7-2/CD86 is constitutively expressed on antigen presenting cells (APCs) such as dendritic cells (DCs), macrophages, and B cells and functions in controlling immune responses through either costimulatory or coinhibitory signals (1,2). Expression of B7-2 is upregulated by APCs upon activation and can be induced in T cells (1,3). The human B7-2 protein, encoded by the CD86 gene, is 329 amino acids (aa) in length with a theoretical molecular weight (MW) of 37.6 kDa (4). The B7-2 protein contains characteristic Ig variable-like (IgV) and constant-like (IgC) domains within its extracellular region (1,3). B7-2/CD86 has structural similarity and shares ~25% sequence homology with another B7 family molecule, B7-1/CD80 (3,5). Both B7-1 and B7-2 are ligands for the activating receptor CD28 and the regulatory receptor cytotoxic T-lymphocyte antigen 4 (CTLA-4) which are expressed on subsets of T cells (1-3,5-7). However, B7-1 and B7-2 bind to CTLA-4 with higher affinity than CD28 (3,5,6). B7-2/CD86 binding to CD28 results in costimulatory signals to promote T cell activation, proliferation, and cytokine production (1-3. 5-7). Binding to CTLA-4 initiates coinhibitory signaling to attenuate the pro-inflammatory T cell response, while also promoting the suppressive function of regulatory T (Treg) cells through expression of indoleamine 2,3-deoxygenase (IDO) (1-3,6,7). Molecules involved in T cell co-signaling have become considerable targets of interest for cancer immunotherapy (7). The anti-CTLA-4 monoclonal antibody ipilimumab, first approved for the treatment of metastatic melanoma, prevents B7-1/B7-2 molecules from binding to CTLA-4, thus driving B7-CD28 binding and promoting costimulatory signals and antitumor effects (1,7,8). Combination therapies targeting co-signaling molecules are currently under investigation to improve antitumor response for treatment of both melanoma and non-melanoma cancers (7,8).

References

1. Collins M, Ling V, Carreno BM. The B7 family of immune-regulatory ligands. Genome Biol. 2005;6(6):223. https://doi.org/10.1186/gb-2005-6-6-223

2. Greaves P, Gribben JG. The role of B7 family molecules in hematologic malignancy. Blood. 2013;121(5):734-744. https://doi.org/10.1182/blood-2012-10-385591

3. Bolandi N, Derakhshani A, Hemmat N, et al. The Positive and Negative Immunoregulatory Role of B7 Family: Promising Novel Targets in Gastric Cancer Treatment. Int J Mol Sci. 2021;22(19):10719. https://doi.org/10.3390/ijms221910719

4. Uniprot (P42081)

5. Bhatia S, Edidin M, Almo SC, Nathenson SG. B7-1 and B7-2: similar costimulatory ligands with different biochemical, oligomeric and signaling properties. Immunol Lett. 2006;104(1-2):70-75. https://doi.org/10.1016/j.imlet.2005.11.019

6. Ohue Y, Nishikawa H. Regulatory T (Treg) cells in cancer: Can Treg cells be a new therapeutic target?. Cancer Sci. 2019;110(7):2080-2089. https://doi.org/10.1111/cas.14069

7. Chen L, Flies DB. Molecular mechanisms of T cell co-stimulation and co-inhibition [published correction appears in Nat Rev Immunol. 2013 Jul;13(7):542]. Nat Rev Immunol. 2013;13(4):227-242. https://doi.org/1010.1038/nri3405

8. Karimi A, Alilou S, Mirzaei HR. Adverse Events Following Administration of Anti-CTLA4 Antibody Ipilimumab. Front Oncol. 2021;11:624780. https://doi.org/101010.3389/fonc.2021.624780

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Antibody Pairs are guaranteed for 6 months from date of receipt.

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Product General Protocols

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Video Protocols

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Blogs on B7-2/CD86.

CD86 - I work in tandem with CD80
CD86 belongs to the immunoglobulin superfamily of proteins that drive innate and adaptive immune responses. It is an 80kD co-stimulatory molecule for the priming and activation of naive and memory T-cells, respectively. CD86 is expressed on activated ...  Read full blog post.

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Bioinformatics

Gene Symbol CD86