Key Product Details

Species Reactivity

Validated:

Rat

Cited:

Mouse, Rat

Applications

Validated:

Immunohistochemistry, Western Blot, ELISA Capture (Matched Antibody Pair), Flow Cytometry, CyTOF-ready

Cited:

Immunohistochemistry, Flow Cytometry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
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Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant rat B7‑2/CD86
Val29-Ile250
Accession # O35531

Specificity

Detects rat B7‑2/CD86 in ELISAs and Western blots. In sandwich immunoassays, less than 20% cross‑reactivity with recombinant mouse (rm) B7-2 is observed and less than 0.2% cross-reactivity with recombinant rat B7-1, rmB7-1, and recombinant human B7-2 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Rat B7‑2/CD86 Antibody

Detection of B7‑2/CD86 in Rat Spleen.

B7‑2/CD86 was detected in immersion fixed paraffin-embedded sections of Rat Spleen using Goat Anti-Rat B7‑2/CD86 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1340) at 10 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface and cytoplasm in splenocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Applications for Rat B7‑2/CD86 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: Rat monocytes

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of Rat Spleen.

Western Blot

0.1 µg/mL
Sample: Recombinant Rat B7‑2/CD86 Fc Chimera (Catalog # 1340-B2)

Rat B7-2/CD86 Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 0.2-0.8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Rat B7‑2/CD86 Biotinylated Antibody (Catalog # BAF1340)
  • Standard: Recombinant Rat B7-2/CD86 Fc Chimera Protein, CF (Catalog # 1340-B2)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: B7-2/CD86

For optimal T cell expansion and activation, a signal induced by the engagement of the T cell receptor and a “co-stimulatory” signal(s) through distinct T cell surface molecules are required. Members of the B7 superfamily of counter-receptors were identified by their ability to interact with co-stimulatory molecules found on the surface of T cells. Members of the B7 superfamily are type I membrane proteins and include B7-1 (CD80), B7-2 (CD86), B7-H1 (PD-L1), B7-H2 (B7RP-1), B7-H3, and PD-L2 (1). B7-2 is expressed constitutively at low levels on most Antigen Presenting Cells (APC) and is rapidly upregulated upon cell activation (2). T cells express two different receptors (CD28 and CTLA-4) capable of binding both B7-1 and B7-2 (2). B7-2 binds to CD28 with the low affinity but binds to CTLA-4 with intermediate affinity. In contrast, B7-1 binds CD28 with intermediate affinity and CTLA-4 with high affinity. Additionally, these molecules have different kinetics for binding CD28 and CTLA-4 with B7-2 having a higher-binding dissociation kinetics (1). Engagement of CD28 by B7-2 increases T cell proliferation and IL-2, IL-4, and IFN-gamma production, thereby enhancing the immune response (3). In contrast, engagement of CTLA-4 is involved in the down-regulation of the immune response (4). Rat B7-2 cDNA encodes a 313 amino acid (aa) precursor protein containing a an extracellular domain, a transmembrane domain, and a cytoplasmic domain. Rat and human B7-1 share 54% aa identity.

References

  1. Coyle, A.J. and J-C. Gutierrez-Ramos (2001) Nature Immunol. 2:203.
  2. Sharpe, A.H. and G.J. Freeman (2002) Nature Reviews 2:116.
  3. Freeman, G.J. et al. (1995) Immunity 5:523.
  4. Walunas, T.L. et al. (1994) Immunity 1:405.

Alternate Names

B72, CD86

Entrez Gene IDs

942 (Human); 12524 (Mouse); 56822 (Rat); 102147235 (Cynomolgus Monkey)

Gene Symbol

CD86

UniProt

Additional B7-2/CD86 Products

Product Documents for Rat B7‑2/CD86 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Rat B7‑2/CD86 Antibody

For research use only

Citations for Rat B7‑2/CD86 Antibody

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Rat B7‑2/CD86 Antibody

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  • Q: I did the WB with CD86 AF1340, I saw two bands, one is like 72KDa, and a stronger band locates higher than 100 KDa and lower than 150KDa. Can you tell me which is which?

    A: The expected MW for rat CD86 would be in the 70 KDa range, depending on glycosylation. We are not certian what the higher size band is because CD86 is not a dimer on the cell surface. However, it is possible that the larger band is a non-specific band that can be removed by increasing the stringency of the western blotting (higher NaCl in the blotting buffer, and increasing the number of washes and concentration of tween-20.  

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