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B7-H3/CD276 Antibody [Unconjugated]

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HAF017). A specific band was detected for B7-H3 at approximately 90-110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." > Western blot shows lysates of LNCaP human prostate cancer cell line and U2OS human osteosarcoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # <a class=HAF017). A specific band was detected for B7-H3 at approximately 90-110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." title="Western blot shows lysates of LNCaP human prostate cancer cell line and U2OS human osteosarcoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for B7-H3 at approximately 90-110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." />
Western blot shows lysates of LNCaP human prostate cancer cell line and U2OS human osteosarcoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody ...read more
AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107)." > AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107)." title="PC-3 human prostate carcinoma cells were stained with Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107)." />
PC-3 human prostate carcinoma cells were stained with Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027, filled histogram) or control antibody (Catalog # ...read more
CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." > CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." title="B7-H3 was detected in immersion fixed paraffin-embedded sections of human melanoma using Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." />
B7-H3 was detected in immersion fixed paraffin-embedded sections of human melanoma using Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) at 15 µg/mL overnight at 4 °C. Tissue was ...read more
HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." > HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." title="Simple Western lane view shows lysates of U2OS human osteosarcoma cell line, loaded at 0.2 mg/mL. A specific band was detected for B7-H3 at approximately 120-160 kDa (as indicated) using 10 µg/mL of Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." />
Simple Western lane view shows lysates of U2OS human osteosarcoma cell line, loaded at 0.2 mg/mL. A specific band was detected for B7-H3 at approximately 120-160 kDa (as indicated) using 10 µg/mL of Goat Anti-Human ...read more
HAF017). A specific band was detected for B7-H3 at approximately 95 kDa (as indicated) in the parental U2OS cell line, but is not detectable in knockout U2OS cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." > HAF017). A specific band was detected for B7-H3 at approximately 95 kDa (as indicated) in the parental U2OS cell line, but is not detectable in knockout U2OS cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." title="Western blot shows lysates of U2OS human osteosarcoma parental cell line and B7-H3 knockout U2OS cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for B7-H3 at approximately 95 kDa (as indicated) in the parental U2OS cell line, but is not detectable in knockout U2OS cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." />
Genetic Strategies: Western blot shows lysates of U2OS human osteosarcoma parental cell line and B7-H3 knockout U2OS cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human B7-H3 Antigen ...read more
CD276 expression in RMS cell lines and PDXs. A CD276 expression levels were evaluated in eleven RMS cell lines, three PDXs, myoblasts, PBMCs, and isolated T cells by WB, revealing high expression in all RMS cell lines, ...read more
B7‐H3 is upregulated in cisplatin resistant neuroblastoma cells and regulates neuroblastoma (NB) cell sensitivity to cisplatin. (A) B7‐H3 mRNA expression in SH‐SY5Y/SH‐SY5Y‐R and SK‐N‐SH/SK‐N‐SH‐R NB ...read more
Expression of CD276 and FGFR4 on RMS-derived xenografts. A Expression of CD276 was assessed by Flow Cytometry on untreated Rh4-derived xenograft tumors and compared to cultured wild-type Rh4 cells by using a ...read more
Expression of CD276 and FGFR4 on RMS-derived xenografts. A Expression of CD276 was assessed by Flow Cytometry on untreated Rh4-derived xenograft tumors and compared to cultured wild-type Rh4 cells by using a ...read more
CD276 expression in RMS cell lines and PDXs. A CD276 expression levels were evaluated in eleven RMS cell lines, three PDXs, myoblasts, PBMCs, and isolated T cells by WB, revealing high expression in all RMS cell lines, ...read more
NUTM2A‐AS1 modulates neuroblastoma (NB) cell sensitivity to cisplatin via B7‐H3. (A) NB cells were transfected with si‐NC, si‐NUTM2A‐AS1#1, and Vector‐B7‐H3 and NUTM2A‐AS1 expression was measured by ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB, Simple Western, Flow, IHC, CyTOF-ready, KO
Clonality
Polyclonal
Host
Goat
Conjugate
Unconjugated
Concentration
LYOPH
Validated by:
     

Genetic Strategies

   

Order Details

B7-H3/CD276 Antibody [Unconjugated] Summary

Immunogen
Mouse myeloma cell line NS0-derived recombinant human B7‑H3
Leu29-Pro245
Accession # NP_079516
Specificity
Detects human B7-H3 in direct ELISAs and Western blots.
Source
N/A
Isotype
IgG
Clonality
Polyclonal
Host
Goat
Gene
CD276
Purity Statement
Antigen Affinity-purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • CyTOF-ready
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunohistochemistry 5-15 ug/mL
  • Knockout Validated
  • Simple Western 10 ug/mL
  • Western Blot 1 ug/mL
Reviewed Applications
Read 1 Review rated 4
using
AF1027 in the following applications:

Publications
Read Publications using
AF1027 in the following applications:

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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Buffer
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.
Preservative
No Preservative
Concentration
LYOPH
Reconstitution Instructions
Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

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

Alternate Names for B7-H3/CD276 Antibody [Unconjugated]

  • 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 regulating immune responses (1‑3). Other family members include B7-1, B7-2, B7-H2, PD-L1 (B7-H1), and PD-L2. B7 proteins are immunoglobulin (Ig) superfamily members with extracellular Ig-V-like and Ig-C-like domains and short cytoplasmic domains. Among the family members, they share about 20‑40% amino acid (aa) sequence identity. The cloned human B7-H3 cDNA encodes a 316 aa type I membrane precursor protein with a putative 28 aa signal peptide, a 217 aa extracellular region containing one V-like and one C-like Ig domain, a transmembrane region, and a 45 aa cytoplasmic domain. An isoform of human B7-H3 containing a four-Ig-like domain extracellular region has also been identified. Human B7-H3 is not expressed on resting B cells, T cells, monocytes or dendritic cells, but is induced on dendritic cells and monocytes by inflammatory cytokines. B7-H3 expression is also detected on various normal tissues and in some tumor cell lines. Human B7-H3 does not bind any known members of the CD28 family of immunoreceptors. However, B7-H3 has been shown to bind an unidentified counter-receptor on activated T cells to costimulate the proliferation of CD4+ or CD8+ T cells. B7-H3 has also been found to enhance the induction of primary cytotoxic T lymphocytes and stimulate IFN-gamma production (1‑3).

  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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Publications for B7-H3/CD276 Antibody (AF1027)(41)

We have publications tested in 4 confirmed species: Human, Mouse, Bovine, Xenograft.

We have publications tested in 9 applications: FACS, Flow Cytometry, ICC, IHC, IHC Paraffin-embedded, IHC-Fr, IHC-P, Immunohistochemistry, Western Blot.


Filter By Application
FACS
(1)
Flow Cytometry
(2)
ICC
(2)
IHC
(10)
IHC Paraffin-embedded
(1)
IHC-Fr
(1)
IHC-P
(6)
Immunohistochemistry
(4)
Western Blot
(10)
All Applications
Filter By Species
Human
(24)
Mouse
(1)
Bovine
(1)
Xenograft
(1)
All Species
Showing Publications 1 - 10 of 41. Show All 41 Publications.
Publications using AF1027 Applications Species
Leo, J;Chen, F;Shi, W;Liang, X;Meng, C;Gu, Q;Albittar, Y;Fan, Z;Zhang, J;Gan, B;Goswami, S;Carmon, K;Frigo, DE;Aparicio, A;Zhao, D; Improve the Efficacy of B7-H3-Targeting Antibody-Drug Conjugate DS-7300a in TP53-deficient Tumors by Inducing Ferroptosis bioRxiv : the preprint server for biology 2026-05-08 [PMID: 42146343] (Human, Xenograft, Immunohistochemistry, Western Blot) Immunohistochemistry, Western Blot Human, Xenograft
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] (IHC, Human) IHC Human
Haydar, D;Ibañez-Vega, J;Crawford, J;Chou, CH;Guy, C;Meehl, M;Yi, Z;Langfitt, D;Vogel, P;DeRenzo, C;Gottschalk, S;Roussel, M;Thomas, P;Krenciute, G; CAR T-cell design dependent remodeling of the brain tumor immune microenvironment identify macrophages as key players that inhibit or promote anti-tumor activity Research square 2023-06-06 [PMID: 37333156] (IHC, Mouse) IHC Mouse
Naohito Hashimoto, Tomonari Suzuki, Keisuke Ishizawa, Sumihito Nobusawa, Hideaki Yokoo, Ryo Nishikawa et al. A clinicopathological analysis of supratentorial ependymoma, ZFTA fusion-positive: utility of immunohistochemical detection of CDKN2A alterations and characteristics of the immune microenvironment Brain Tumor Pathology 2023-07-01 [PMID: 37322295]
Xiaolei Shi, Abderrahman Day, Hannah E. Bergom, Sydney Tape, Sylvan C. Baca, Zoi E. Sychev et al. Integrative molecular analyses define correlates of high B7-H3 expression in metastatic castrate-resistant prostate cancer npj Precision Oncology 2022-11-02 [PMID: 36323882]
Stine Asferg Petterson, Mia Dahl Sørensen, Mark Burton, Mads Thomassen, Torben A. Kruse, Signe Regner Michaelsen et al. Differential expression of checkpoint markers in the normoxic and hypoxic microenvironment of glioblastomas Brain Pathology 2023-01-01 [PMID: 36093941]
J Joung, PC Kirchgatte, A Singh, JH Cho, SP Nety, RC Larson, RK Macrae, R Deasy, YY Tseng, MV Maus, F Zhang CRISPR activation screen identifies BCL-2 proteins and B3GNT2 as drivers of cancer resistance to T cell-mediated cytotoxicity Nature Communications, 2022-03-25;13(1):1606. 2022-03-25 [PMID: 35338135] (Western Blot, Human) Western Blot Human
Mengli Zhang, Huaichao Zhang, Minjie Fu, Jingwen Zhang, Cheng Zhang, Yingying Lv et al. The Inhibition of B7H3 by 2-HG Accumulation Is Associated With Downregulation of VEGFA in IDH Mutated Gliomas Frontiers in Cell and Developmental Biology 2021-05-17 [PMID: 34079802]
R Wang, L Sun, S Xia, H Wu, Y Ma, S Zhan, G Zhang, X Zhang, T Shi, W Chen B7-H3 suppresses doxorubicin-induced senescence-like growth arrest in colorectal cancer through the AKT/TM4SF1/SIRT1 pathway Cell Death & Disease, 2021-05-06;12(5):453. 2021-05-06 [PMID: 33958586] (IHC, Human) IHC Human
Myungeun Lee, Zhi Hong Lu, Charles B. Shoemaker, Jacqueline M. Tremblay, Bradley St. St. Croix, Steven Seaman et al. Advanced genetic engineering to achieve in vivo targeting of adenovirus utilizing camelid single domain antibody Journal of Controlled Release 2021-06-01 [PMID: 33872627]
Show All 41 Publications.

Review for B7-H3/CD276 Antibody (AF1027) (1) 41

Average Rating: 4
(Based on 1 review)
We have 1 review tested in 1 species: Human.

Reviews using AF1027:
Filter by Applications
IHC-Fr
(1)
All Applications
Filter by Species
Human
(1)
All Species
Images Ratings Applications Species Date Details
  4
reviewed by:
Verified Customer
IHC-Fr Human 01/05/2015
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Summary

ApplicationImmunohistochemistry-Frozen
Sample TestedSee PMID 22080438
SpeciesHuman

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Verified Customer
01/05/2015
Application: IHC-Fr
Species: Human

Bioinformatics

Gene Symbol CD276
Uniprot