B7-2/CD86 Antibody (BU63) - Azide and BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-34569

Clone BU63 was used by HLDA to establish CD designation.
Novus Biologicals, part of Bio-Techne
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NBP2-34569-0.1mg
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Human, Mouse, Rat

Applications

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

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # BU63

Format

Azide and BSA Free

Concentration

1.0 mg/ml

View all B7-2/CD86 Antibodies »

Product Specifications

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

1.0 mg/ml of antibody purified from Bioreactor Concentrate by Protein A/G. Prepared in 10mM PBS WITHOUT BSA & azide. Also available at 200 ug/ml WITH BSA & azide (NBP2-25208).

Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80C.

Scientific Data Images for B7-2/CD86 Antibody (BU63) - Azide and BSA Free

Knockout Validated: B7-2/CD86 Antibody (BU63) - Azide and BSA Free [NBP2-34569]

Knockout Validated: B7-2/CD86 Antibody (BU63) - Azide and BSA Free [NBP2-34569]

Knockout Validated: B7-2/CD86 Antibody (BU63) - Azide and BSA Free [NBP2-34569] - 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 # NBP2-34569) 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.
Immunocytochemistry/ Immunofluorescence: B7-2/CD86 Antibody (BU63) - Azide and BSA Free [NBP2-34569]

Immunocytochemistry/ Immunofluorescence: B7-2/CD86 Antibody (BU63) - Azide and BSA Free [NBP2-34569]

Immunocytochemistry/Immunofluorescence: B7-2/CD86 Antibody (BU63) - Azide and BSA Free [NBP2-34569] - Immunofluorescence staining of PFA-fixed Ramos cells using B7-2/CD86 Antibody (BU63) followed by goat anti-Mouse IgG conjugated to CF488 (green). Nuclei are stained with Red Dot.
Immunohistochemistry-Paraffin: B7-2/CD86 Antibody (BU63) - Azide and BSA Free [NBP2-34569]

Immunohistochemistry-Paraffin: B7-2/CD86 Antibody (BU63) - Azide and BSA Free [NBP2-34569]

Immunohistochemistry-Paraffin: B7-2/CD86 Antibody (BU63) - Azide and BSA Free [NBP2-34569] - Formalin-fixed, paraffin-embedded esophagus tumor tissue stained with CD86 antibody (5 ug/ml), peroxidase-conjugate and DAB chromogen. Note strong staining of differentiated squamous cells. TMA was used for this test.
View 2 more images

Applications for B7-2/CD86 Antibody (BU63) - Azide and BSA Free

Application
Recommended Usage

Flow Cytometry

1-2ug/million cells

Immunocytochemistry/ Immunofluorescence

1-2ug/ml

Immunohistochemistry-Frozen

0.5-1ug/ml

Immunohistochemistry-Paraffin

2-4ug/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)

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

10 mM PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1.0 mg/ml

Shipping

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

Stability & Storage

Store at -20 to -80C. Avoid freeze-thaw cycles.

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

Additional B7-2/CD86 Products

Product Documents for B7-2/CD86 Antibody (BU63) - Azide and BSA Free

Product Datasheets

Certificate of Analysis

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Product Specific Notices for B7-2/CD86 Antibody (BU63) - Azide and BSA Free

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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