Anti-Human B7-1

Product Datasheet

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Anti-Human B7-1

Purified No Carrier Protein

(CD80)

Monoclonal Antibody

Prod. No.:	B566
Clone:	37711
Isotype:	Mouse IgG₁
Pkg. Size:	1 mg, 500 µg
Storage:	-20°C to -70°C Detailed storage instructions below.

Description

Specificity:

Mouse Anti-Human B7-1 (Clone 37711) recognizes an epitope on Human B7-1. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype.

Background:

It is widely accepted that T cells require a signal induced by the engagement of the T cell receptor and a “costimulatory” signal(s) through distinct T cell surface molecules for optimal T cell expansion and activation. Members of the B7 superfamily of counterreceptors were identified by their ability to interact with costimulatory molecules found on the surface of T cells. Members of the B7 superfamily include B7-1 (CD80), B7-2 (CD86), B7-H1 (PD-L1), B7-H2 (B7RP-1), B7-H3 and PD-L2. B7-1 is expressed on activated B cells, activated T cells, mature dendritic cells and macrophages. T cells express two different receptors capable of binding B7-1, CD28 and CTLA-4. CTLA-4 binds to B7-1 with a 20 - 100 fold higher affinity than CD28. Engagement of CD28 by B7-1 increases T cell proliferation and IL-2 production and enhances the immune response. In contrast, engagement of CTLA-4 is involved in the down-regulation of the immune response. Rat and human B7-1 share 47% amino acid identity.

Known Species Cross Reactivity: Hu+

Host Species

Mouse

Immunogen

Purified Recombinant Human B7-1 (>98%)

Formulation

This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present.

Reconstitution

This lyophilized antibody may be aseptically reconstituted with sterile PBS to a final volume of 1 ml. This will result in a final antibody concentration of 500 µg/ml.

Storage and Stability

The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months.

Reported Applications

ELISA Cap, IHC (NBF/Par.), FC, N

ELISA Sandwich: This antibody is useful as the capture antibody in a sandwich ELISA. The suggested coating concentration is 4 µg/ml. A suitable detection antibody is Anti-Mouse B7-1-Biotin (Antigen Aff. Pur.) (Leinco Prod. No.: B394). When used in conjunction with an optimal second step reagent such as UltraAvidin™- Horseradish Peroxidase (HRPO) (Leinco Prod. No.: A106) and a chromogenic substrate such as TMB HRPO Microwell Substrate Standard Kinetic One Component "Ready Use" (Leinco Prod. No.: T118), the sensitivity of this sandwich ELISA is 4 ng/well. A suggested standard for this assay is Recombinant Human B7-1 (Leinco Prod. No.: B532).

IHC (NBF/Par.): This antibody should give satisfactory staining results when used at a concentration of 25 µg/ml. The recommended secondary antibody for IHC is Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114). For chromogenic detection with high signal and low background use 3, 3'-Diaminobenzidine Tetrahydrochloride (DAB) Substrate Kit (Leinco Prod. No.: D100) or Optional Metal Enhanced DAB Substrate Kit (Leinco Prod. No.: K107).

Flow Cytometry: It is recommended to use the indirect method for signal enhancement when enumerating cells expressing B7-1. A suggested method would be to stain cells expressing B7-1 with 0.25 µg per 1-2.5 x 10⁵ cells in a total staining volume of ≤200 µl followed by Goat Anti-Mouse IgG (H&L)-R-phycoerythrin (Leinco Prod. No.: M1189).

Neutralization: This antibody is useful for neutralization of Human B7-1 bioactivity. The antibody dose required to neutralize 50% (ND₅₀) of the biological activity of Human B7-1 (at 0.1 µg/ml) is 0.001 - 0.002 µg/ml.

Each investigator should determine their own optimal working dilution for specific applications.

Country of Origin

USA