Anti-Mouse B7-H1 (Antigen Aff. Pur.)

Product Datasheet

www.leinco.com

excellence in early discovery research

Goat Anti-Mouse B7-Homology 1 (B7-H1)

Antigen Affinity Purified

(B7-H, MGC142294, MGC142296, PD-L1, PDCD1L1, PDCD1LG1, PDL1)

Polyclonal Antibody

Prod. No.:	B557
Host:	Goat
Pkg. Size:	1 mg, 100 µg
Storage:	-20°C to -70°C Detailed storage instructions below.

Description

Specificity:

Goat Anti-Mouse B7-H1 recognizes Mouse B7-H1. 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 homolog 1 (B7-H1), also called programmed cell death 1 ligand 1 (PDCD1L1) and programmed death ligand 1 (PDL1), is a member of the growing B7 family of immune proteins that provide signals for both stimulating and inhibiting T cell activation. Other family members include B7-1, B7-2, B7-H2, PDL2 and B7-H3. B7 proteins are members of the immunoglobulin (Ig) superfamily, their 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-H1 share approximately 70% amino acid sequence identity. B7-H1 has been identified as one of two ligands for programmed death-1 (PD-1), a member of the CD28 family of immunoreceptors.

B7-H1 is found in a variety of non-lympoid parenchymal organs, including the heart, placenta, skeletal muscle and lung and has also been detected in most human cancers. On previously activated T cells, B7-H1 interaction with PD-1 inhibits TCR-mediated proliferation and cytokine production, suggesting an inhibitory role in regulating immune responses. However, a costimulatory function for the PD-1 ligands on resting T cells has also been reported.

Known Species Cross Reactivity: Ms+

Immunogen

NSO-Derived Recombinant Mouse B7-H1 Extracellular Domain Specific Goat IgG

Formulation

This 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 5.0% w/v trehalose with no calcium, magnesium, or preservatives present.

Reconstitution

We suggest aseptically reconstituting this lyophilized, antigen affinity purified polyclonal antibody with 1.0 ml of sterile 1X PBS pH 7.2-7.3. This will result in a final antibody concentration of 0.1 mg/ml.

Endotoxin

<1.0 EU/µg as determined by the LAL method

Storage and Stability

The lyophilized antigen affinity purified polyclonal antibody can be stored desiccated at -20°C to -70°C for twelve months from date of receipt. 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

Indirect ELISA, IHC (NBF/Par.), FC, WB

Indirect ELISA: This antibody can be used in an indirect detection ELISA at 0.5 - 1.0 µg/ml with a suitable second step reagent such as Donkey Anti-Goat IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: G505). The detection sensitivity of this indirect ELISA for Mouse B7-H1 is approximately 0.3 ng/well. In this format, this antibody shows approximately 20% cross-reactivity with rhB7-H1.

IHC (NBF/Par.): This antibody should give satisfactory staining results when used at a concentration of 2-15 µg/ml. The recommended secondary antibody for IHC is Donkey Anti-Goat IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: G505). 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-H1. A suggested method would be to stain cells expressing B7-H1 with 3-10 µg per 1 x 10⁶ cells in a total staining volume of 100 µl followed by Donkey F(ab')₂ Anti-Goat IgG (H&L)-R-phycoerythrin (R-PE) (Leinco Prod. No.: G641).

Western Blotting: To detect Mouse B7-H1 this polyclonal antibody can be used at a concentration of 0.1-0.2 µg/ml. This polyclonal antibody should be used in conjunction with compatible second-step reagents such as Donkey Anti-Goat IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: G505) and a chromogenic substrate such as TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343). The detection limit for Mouse B7-H1 is 5 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 Mouse B7-H1 (Leinco Prod. No.: B526).

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

Country of Origin

USA