B7-2/CD86 Antibody (BU63)

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-25208

Clone BU63 was used by HLDA to establish CD designation.
Novus Biologicals, part of Bio-Techne
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NBP2-25208-0.02mg
NBP2-25208
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Key Product Details

Validated by

Independent Antibodies

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Flow Cytometry, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Knockout Validated, Western Blot

Cited:

Block/Neutralize, IF/IHC, Immunocytochemistry/ Immunofluorescence, Immunofluorescence, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # BU63

Concentration

0.2 mg/ml

View all B7-2/CD86 Antibodies »

Product Summary for B7-2/CD86 Antibody (BU63)

Immunogen

ARH-77 (B-lymphoblastoid cell line)

Localization

Cell Surface

Marker

Dendritic Cells Maturation Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Theoretical MW

70 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Description

200ug/ml of antibody purified from Bioreactor Concentrate by Protein A or G. Prepared in 10 mM PBS with 0.05% BSA & 0.05% azide. Also available WITHOUT BSA & azide at 1.0 mg/ml. (NBP2-34569)

Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80C. Antibody is stable for 24 months. Non-hazardous.

Scientific Data Images for B7-2/CD86 Antibody (BU63)

Knockout Validated: B7-2/CD86 Antibody (BU63) [NBP2-25208] - Western blot shows lysates of Ramos human Burkitt's lymphoma parental cell line and B7-2 knockout (KO) Ramos cell line. PVDF membrane was probed with 1.0 ug/mL of Mouse Anti-Human B7-2 Monoclonal Antibody (Catalog # NBP2-25208) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog #HAF018). Specific band was detected for B7-2 at approximately 90 kDa (as indicated) in the parental Ramos cell line, but is not detectable in the knockout Ramos cell line. This experiment was conducted under reducing conditions.
Immunohistochemistry: B7-2/CD86 Antibody (BU63) [NBP2-25208] - Macrophage polarization states. Distribution and proportion of CD86+ macrophages (brown) were identified using IHC procedures. Intensity for the color response to antibodies was visible in lung tissues from all groups. Population proportions for the stained cells as shown by red arrows were calculated as a fold change of the control. (d). The results were expressed as Mean +/- SD (n = 3). **: a p-value of < 0.01 vs NS, OVA/BUD and OVA/PCI except the number of CD86+ cells in OVA/PCI group. #: P < 0.05 vs NS and ##; P < 0.01 vs either NS or OVA/BUD group. Image collected and cropped by CiteAb from the following publication (https://respiratory-research.biomedcentral.com/articles/10.1186/s12931-020-1322-5) licensed under a CC-BY license.
Immunocytochemistry/Immunofluorescence: B7-2/CD86 Antibody (BU63) [NBP2-25208] - Immunofluorescence staining of PFA-fixed Ramos cells using followed by goat anti-Mouse IgG conjugated to CF488 (green). Nuclei are stained with Red Dot.
View 4 more images

Applications for B7-2/CD86 Antibody (BU63)

Application
Recommended Usage

Flow Cytometry

1-2 ug/million cells

Immunocytochemistry/Immunofluorescence

1-2 ug/ml

Immunohistochemistry

0.5-1ug/ml

Immunohistochemistry-Paraffin

2-4 ug/ml

Western Blot

0.5-1ug/ml
Application Notes
Immunohistochemistry (Formalin-fixed): 2-4ug/ml for 30 minutes at RT. Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris buffer with 1mM EDTA, pH 9.0, for 45 min at 95C followed by cooling at RT for 20 minutes.
Optimal dilution for a specific application should be determined.
IHC-fr reported in the literature (PMID: 29867472, 31629891)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

10 mM PBS with 0.05% BSA

Preservative

0.05% Sodium Azide

Concentration

0.2 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C.

Background: B7-2/CD86

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

Alternate Names

B72, CD86

Gene Symbol

CD86

UniProt

P42081 (Human)

Additional B7-2/CD86 Products

Product Documents for B7-2/CD86 Antibody (BU63)

Product Datasheets

Certificate of Analysis

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Product Specific Notices for B7-2/CD86 Antibody (BU63)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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