Rat B7-2/CD86 Biotinylated Antibody

Catalog # Availability Size / Price Qty
BAF1340
Detection of B7‑2/CD86 in Rat Spleen.
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Rat B7-2/CD86 Biotinylated Antibody Summary

Species Reactivity
Rat
Specificity
Detects rat B7‑2/CD86 in ELISAs and Western blots. In sandwich immunoassays, approximately 50% cross-reactivity with recombinant mouse (rm) B7‑2 is observed, 10% cross-reactivity with recombinant human B7-2 is observed, and less than 1% cross-reactivity with rmB7-1, rmB7‑H1, rmB7-H3, and rmPD-L2 is observed.
Source
Polyclonal Goat IgG
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Label
Biotin

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Rat B7‑2/CD86 Fc Chimera (Catalog # 1340-B2)
Immunohistochemistry
5-15 µg/mL
Immersion fixed paraffin-embedded sections of Rat Spleen.

Rat B7-2/CD86 Sandwich Immunoassay

Recommended Concentration
Reagent
ELISA Detection (Matched Antibody Pair)
0.1-0.4 µg/mL 

Use in combination with:

Capture Reagent: Rat B7‑2/CD86 Antibody (Catalog # AF1340)

Standard: Recombinant Rat B7-2/CD86 Fc Chimera Protein, CF (Catalog # 1340-B2)

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunohistochemistry View Larger

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 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF1340) 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.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

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.
Entrez Gene IDs
942 (Human); 12524 (Mouse); 56822 (Rat); 102147235 (Cynomolgus Monkey)
Alternate Names
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

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