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Mouse B7-H3 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF1397

R&D Systems, part of Bio-Techne
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AF1397
AF1397-SP

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Mouse

Applications

Validated:

Neutralization, Western Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse B7-H3
Val29-Phe244
Accession # Q8VE98

Specificity

Detects mouse B7‑H3 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 50% cross‑reactivity with recombinant human (rh) B7-H3 is observed, and less than 1% cross-reactivity with recombinant mouse (rm) B7‑H1, rmB7-H2 and rmB7-1 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Mouse B7-H3 Antibody

Detection of Mouse B7-H3 antibody by Western Blot.

Detection of Mouse B7‑H3 by Western Blot.

Western blot shows lysates of MEF mouse embryonic feeder cells, P19 mouse embryonal carcinoma cell line, NIH-3T3 mouse embryonic fibroblast cell line, C2C12 mouse myoblast cell line, and 3T3-L1 mouse embryonic fibroblast adipose-like cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1397) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for B7-H3 at approximately 45-55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Proliferation Induced by B7‑H3 and Neutralization by Mouse B7‑H3 Antibody.

Proliferation Induced by B7‑H3 and Neutralization by Mouse B7‑H3 Antibody.

Recombinant Mouse B7-H3 enhances proliferation in mouse CD3+T cells in the presence of 100 ng/mL Hamster Anti-Mouse CD3e Monoclonal Antibody (Catalog # MAB484) in a dose-dependent manner (orange line), as measured by the Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Mouse B7-H3 (2 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1397). The ND50 is typically 1.5-7.5 µg/mL.

Applications for Mouse B7-H3 Antibody

Application
Recommended Usage

Western Blot

1 µg/mL
Sample: MEF mouse embryonic feeder cells, P19 mouse embryonal carcinoma cell line, NIH‑3T3 mouse embryonic fibroblast cell line, C2C12 mouse myoblast cell line, and 3T3‑L1 mouse embryonic fibroblast adipose-like cell line

Neutralization

Measured by its ability to neutralize B7‑H3-induced proliferation in mouse CD3+ T cells. The Neutralization Dose (ND50) is typically 1.5-7.5 µg/mL in the presence of 2 µg/well Recombinant Mouse B7‑H3 and 100 ng/mL Hamster Anti-Mouse CD3 epsilon Monoclonal Antibody (Catalog # MAB484).
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: B7-H3

T cells require a signal induced by the engagement of the T cell receptor and a “co‑stimulatory” signal(s) through distinct T cell surface molecules for optimal T cell expansion and activation. Members of the B7 superfamily of counter-receptors were identified by their ability to interact with co‑stimulatory 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 (1). B7-H3 is expressed at very high levels in immature dendritic cells at moderate levels on mature dendritic cells, LPS stimulated immature dendritic cells and LPS stimulated monocytes, and at low levels on resting monocytes. B7-H3 binds to activated T cells via an as-of-yet identified receptor. B7-H3 co-stimulates proliferation of T cells and interferon-gamma (IFN-gamma) production and enhances the induction of cytotoxic T cells. B7-H3 shares 20‑27% amino acid (aa) identity with other B7 family members (2). Murine B7-H3 is a 259 aa protein containing an extracellular domain, a transmembrane domain and a cytoplasmic domain. Mouse and human B7-H3 share 87% aa identity (3).

References

  1. Coyle, A.J. and J.-C. Gutierrez-Ramos (2001) Nature Immunol. 2:203.
  2. Chapoval, A.I. et al. (2001) Nature Immunol. 2:269.
  3. Sun, M. et al. (2002) J. Immunol. 168:6294.

Long Name

B7 Homolog 3

Alternate Names

B7H3, CD276

Entrez Gene IDs

80381 (Human); 102657 (Mouse); 315716 (Rat); 102131295 (Cynomolgus Monkey)

Gene Symbol

CD276

UniProt

Additional B7-H3 Products

Product Documents for Mouse B7-H3 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse B7-H3 Antibody

For research use only

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