B7-2/CD86 Antibody (SJ20-00)

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-67417

Recombinant Monoclonal Antibody.
Novus Biologicals, part of Bio-Techne
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NBP2-67417
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Rat

Applications

Validated:

Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Western Blot

Cited:

IF/IHC, Immunohistochemistry-Paraffin

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # SJ20-00

Concentration

1 mg/ml

View all B7-2/CD86 Antibodies »

Product Specifications

Immunogen

Synthetic peptide within Human B7-2/CD86 aa 1-50 / 329. (SwissProt: P42081 Human; Entrez Gene:56822 Rat)

Localization

Cell membrane.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for B7-2/CD86 Antibody (SJ20-00)

B7-2/CD86 Antibody (SJ20-00)

Immunocytochemistry/Immunofluorescence: B7-2/CD86 Antibody (SJ20-00) [NBP2-67417] -

Immunocytochemistry/Immunofluorescence: B7-2/CD86 Antibody (SJ20-00) [NBP2-67417] - Analysis of Raji cells labeling CD86 with Rabbit anti-CD86 antibody at 1/500 dilution.Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD86 antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594,was used as the secondary antibody at 1/1,000 dilution.
B7-2/CD86 Antibody (SJ20-00)

Western Blot: B7-2/CD86 Antibody (SJ20-00) [NBP2-67417] -

Western Blot: B7-2/CD86 Antibody (SJ20-00) [NBP2-67417] - Analysis of CD86 on different lysates with Rabbit anti-CD86 antibody at 1/1,000 dilution.Lane 1: Raji cell lysate (10 ug/Lane)Lane 2: Mouse lung tissue lysate (20 ug/Lane)Lane 3: Rat lung tissue lysate (20 ug/Lane)Predicted band size: 38 kDaObserved band size: 70 kDaExposure time: 1 minute;4-20% SDS-PAGE gel.Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.
B7-2/CD86 Antibody (SJ20-00)

Western Blot: B7-2/CD86 Antibody (SJ20-00) [NBP2-67417] -

Western Blot: B7-2/CD86 Antibody (SJ20-00) [NBP2-67417] - Analysis of CD86 on different lysates with Rabbit anti-CD86 antibody at 1/2,000 dilution.Lane 1: Daudi cell lysateLane 2: Raji cell lysateLane 3: Ramos cell lysateLane 4: Jurkat cell lysateLane 5: K-562 cell lysateLysates/proteins at 15 ug/Lane.Predicted band size: 38 kDaObserved band size: 70 kDaExposure time: 2 minutes 37 seconds;4-20% SDS-PAGE gel.Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.

Applications for B7-2/CD86 Antibody (SJ20-00)

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:500

Immunoprecipitation

Use at an assay dependent concentration

Western Blot

1:500-1:2000
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

TBS (pH7.4), 0.05% BSA, 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 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 short term. Aliquot and store at -20C long term. 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

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Product Specific Notices for B7-2/CD86 Antibody (SJ20-00)

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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