Recombinant Human B7-H3 (4Ig)/B7-H3b Protein

• Reactivity: Hu
• Applications: Bioactivity

Recombinant Human B7-H3 (4Ig)/B7-H3b Protein Summary

• Details of Functionality: Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated human T cells. T lymphocytes cultured for 72 hours with PHA were incubated for an additional 3 days in 96 well plates coated with 500 ng/mL anti-CD3 and/or 10 µg/mL rhB7-H3 (4Ig)/B7-H3b.
  The presence of rhB7-H3 (4Ig)/B7-H3b at 10 µg/mL inhibited the anti-CD3 response by >30%.
  Optimal dilutions should be determined by each laboratory for each application.
• Source: Mouse myeloma cell line, NS0-derived human B7-H3 protein
  Gly27-Thr461, with a C-terminal 10-His tag
• Accession #: Q5ZPR3
• N-terminal Sequence: Gly27
• Protein/Peptide Type: Recombinant Proteins
• Gene: CD276
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 48 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 78-90 kDa, reducing conditions

Packaging, Storage & Formulations

• 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Reconstitution Instructions: Reconstitute at 100-200 μg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human B7-H3 (4Ig)/B7-H3b Protein

• B7H3
• B7-H3
• B7H34Ig-B7-H3
• B7-H3B7 homolog 3
• CD276 antigen
• CD276 molecule
• CD276
• Costimulatory molecule

Background

Human B7 homolog 3 (B7-H3) is a member of the B7 family of immune proteins that provide signals for the regulation of immune responses (1 - 3). Other family members include B7-1, B7-2, B7-H1/PD-L1, B7-H2, and PD-L2. B7 family proteins are type I transmembrane immunoglobulin (Ig) superfamily members that contain extracellular Ig V‑like and Ig C‑like domains with a short cytoplasmic tail. Among the family members there is about 20 - 40% amino acid (aa) sequence identity. B7-H3 was initially reported to be a 316 aa type I transmembrane precursor protein that contained a signal sequence, an extracellular region with one V‑type and one C‑type Ig domain, a transmembrane segment and a short cytoplasmic tail (1). Subsequent studies have identified a second 110 kDa form whose precursor is 534 aa in length. Termed 4IgB7-H3 or B7-H3b, this molecule has two additional Ig-like domains (one V‑type and one C‑type) and shows a ubiquituous expression pattern (4, 5). It would appear that the human 4Ig form is the principal, if not the only form of B7-H3 (5). Its precursor contains a 26 aa signal sequence, a 435 aa extracellular region, a 31 aa transmembrane domain, and a 42 aa cytoplasmic tail. The four Ig-like domains alternate between V‑type and C‑type, and apparently are the consequence of a V‑C type tandem duplication (4, 5). B7-H3b is expressed on dendritic cells as well as activated T, B and NK cells (5). The mouse gene differs from that of human in that it cannot code for four Ig-like domains; only a V‑type:C‑type pair (4). Human B7-H3b binding to an undefined receptor has shown to be inhibitory to NK cell illing and cytokine release (6). It also seems to be required for late stage osteoblast differentiation (7).

1. Chapoval, A.I. et al. (2001) Nat. Immunol. 2:269.
2. Sharpe, A.H. and G.J. Freeman (2002) Nat. Rev. Immunol. 2:116.
3. Coyle, A. and J.Gutierrez-Ramos (2001) Nat. Immunol. 2:203.
4. Sun, M. et al. (2002) J. Immunol. 168:6294.
5. Steinberger, P. et al. (2004) J. Immunol. 172:2352.
6. Prasad, D.V.R. et al. (2004) J. Immunol. 173:2500.
7. Suh, W-K. et al. (2004) Proc. Natl. Acad. Sci. USA 101:12969.

Publications for B7-H3/CD276 (2318-B3)(4)

Publications using 2318-B3

• Li, D;Wang, R;Liang, T;Ren, H;Park, C;Tai, CH;Ni, W;Zhou, J;Mackay, S;Edmondson, E;Khan, J;Croix, BS;Ho, M; Camel nanobody-based B7-H3 CAR-T cells show high efficacy against large solid tumours Nature communications 2023-09-22 [PMID: 37739951] (Bioassay, N/A)
• Seo, YR;Lee, J;Ryu, HS;Kim, EG;Kim, SH;Jeong, J;Jung, H;Jung, Y;Kim, HB;Jo, YH;Kim, YD;Jin, MS;Lee, YY;Kim, KM;Yi, EC; Lateral interactions between CD276 and CD147 are essential for stemness in breast cancer: a novel insight from proximal proteome analysis Scientific reports 2023-08-30 [PMID: 37648771] (Bioassay, Human)
• H Li, Z Yang, X Yang, F Zhang, J Wang, Z Wu, C Wanyan, Q Meng, W Gao, X Yang, J Wei LINC01123 promotes immune escape by sponging miR-214-3p to regulate B7-H3 in head and neck squamous-cell carcinoma Cell Death & Disease, 2022-02-03;13(2):109. 2022-02-03 [PMID: 35115487] (ELISA Capture, Human)
• BR Yoon, YH Chung, SJ Yoo, K Kawara, J Kim, IS Yoo, CG Park, SW Kang, WW Lee Preferential Induction of the T Cell Auxiliary Signaling Molecule B7-H3 on Synovial Monocytes in Rheumatoid Arthritis J. Biol. Chem, 2015-12-23;291(8):4048-57. 2015-12-23 [PMID: 26702052] (Bioassay, Human)

Bioinformatics

• Gene Symbol: CD276
• Uniprot:
  • Q5ZPR3()