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Goat Anti-Human B7 Homolog 2 (B7-H2)

Biotin

(B7RP-1, B7h, LICOS, GL50)

Polyclonal Antibody

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Prod. No.:	B607
Host:	Goat
Pkg. Size:	50 µg, 1 mg
Storage:	-20°C to -70°C Detailed storage instructions below.

Description

Specificity:

Goat Anti-Human B7-Homolog 2 (B7-H2) recognizes Human B7-H2. This antigen affinity purified polyclonal antibody was purified using a proprietary chromatographic technique that includes covalently immobilizing the antigen proteins or peptides to agarose based beads. This purification method enhances specificity, reduces nonspecific binding of extraneous IgG and provides you with the most reliable reagent available for your early discovery research.

Background:

Human B7-H2, also called B7RP-1, B7h, LICOS, and GL50, is a member of the growing B7 family of immune costimulatory proteins. Other family members include B7-1, B7-2, B7-H1 (PD-L1), PD-L2, and B7-H3. B7 proteins are members of the immunoglobulin (Ig) superfamily, the extracellular domains contain 2 Ig-like domains and all members have short cytoplasmic domains. Among the family members, they share about 20 - 25% amino acid identity. Human and mouse B7-H2 share approximately 49% amino acid identity. B7-H2 has been identified as the ligand for ICOS, a member of the CD28 family of costimulatory receptors. The B7-H2/ICOS interaction appears to play roles in T cell dependent B cell activation and Th differentiation.

Known Species Cross Reactivity: Hu+

Immunogen

Purified Recombinant Human B7-H2 (>98%)

Formulation

This biotinylated antigen affinity purified polyclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco’s phosphate buffered saline (1X PBS) pH 7.2 – 7.3 containing 50 µg of bovine serum albumin per µg of antibody with no calcium, magnesium, or preservatives present.

Reconstitution

We suggest aseptically reconstituting this biotinylated, lyophilized, antigen affinity purified polyclonal antibody with 1 ml of sterile 20 mM Trizma base, 0.15 M sodium chloride (1X Tris Buffered Saline) pH 7.2-7.3, containing 0.1% w/v bovine serum albumin (BSA). This will result in a final biotinylated antibody concentration of 50 µg/ml.

Storage and Stability

The lyophilized, biotinylated antigen affinity purified polyclonal antibody can be stored desiccated at -20°C to -70°C for up to twelve months from date of receipt. The reconstituted bioin conjugate can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted conjugate, 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

FC, WB

Flow Cytometry: It is recommended to use the indirect method for signal enhancement when enumerating cells expressing B7-H2. A suggested method would be to stain cells expressing B7-H2 with 25 µg per 1 - 2.5 x 10⁵ cells in a total staining volume of 200 µl followed by UltraAvidin™- R-phycoerythrin (R-PE) (Leinco Prod. No.: A104).

Western Blotting: To detect Human B7-H2 this biotin conjugate can be used at a concentration of 0.1 - 0.2 µg/ml. This biotin conjugate should be used in conjunction with compatible second-step reagents such as UltraAvidin™- Horseradish Peroxidase (HRPO) (Leinco Prod. No.: A106) and a chromogenic substrate such as TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343). The detection limit for Human B7-H2 is 1 ng/lane under either reducing or non-reducing conditions. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used. A suitable Western blotting control is Recombinant Human B7-H2 (Leinco Prod. No.: B537).

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

Country of Origin

USA