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?

Name: Rat B7-2/CD86 Antibody
Brand: R&D Systems
SKU: AF1340
Price: 469 USD

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.

Rat B7-2/CD86 Antibody

Catalog #: AF1340
3 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF1340
AF1340-SP

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All B7-2/CD86 Antibodies

Product Details

Citations (3)

FAQs

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Rat B7-2/CD86 Antibody Summary

Species Reactivity

Rat

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.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

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

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Rat B7‑2/CD86 Fc Chimera (Catalog # 1340-B2)

Flow Cytometry

2.5 µg/10⁶ cells

Rat monocytes

Immunohistochemistry

5-15 µg/mL

Immersion fixed paraffin-embedded sections of Rat Spleen.

CyTOF-ready

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

Rat B7-2/CD86 Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

0.2-0.8 µg/mL

Use in combination with:

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

Scientific Data

Immunohistochemistry

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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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile 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.

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

Coyle, A.J. and J-C. Gutierrez-Ramos (2001) Nature Immunol. 2:203.
Sharpe, A.H. and G.J. Freeman (2002) Nature Reviews 2:116.
Freeman, G.J. et al. (1995) Immunity 5:523.
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

Product Datasheets

Product Datasheet

COA

SDS

Related Research Areas

Citations for Rat B7-2/CD86 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Synthetic polymer coatings diminish chronic inflammation risk in large ECM-based materials
Authors: Laura G. Bracaglia, Shira Winston, Douglas A. Powell, John P. Fisher
Journal of Biomedical Materials Research Part A
Bone marrow mesenchymal stem cell-derived exosomal microRNA-125a promotes M2 macrophage polarization in spinal cord injury by downregulating IRF5
Authors: Q Chang, Y Hao, Y Wang, Y Zhou, H Zhuo, G Zhao
Brain research bulletin, 2021-02-17;170(0):199-210.
Species: Mouse
Sample Types: Whole Cells
Applications: Flow Cytometry
Anti-inflammatory effects of new human histamine H3 receptor ligands with flavonoid structure on BV-2 neuroinflammation
Authors: Ewelina Honkisz-Orzechowska, Katarzyna Popiołek-Barczyk, Zuzanna Linart, Jadwiga Filipek-Gorzała, Anna Rudnicka, Agata Siwek et al.
Inflammation Research

FAQs

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?
- 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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