B7-H3/CD276 Monoclonal antibody

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  WB analysis of HEK-293 using 66481-1-Ig

  HEK-293, PC-3, LNCaP, HeLa cells, and human spleen, human placenta, pig spleen tissues were subjected to SDS PAGE followed by western blot with 66481-1-Ig (B7-H3/CD276 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 66481-1-Ig

  WB result of B7-H3/CD276 antibody (66481-1-Ig; 1:40000; incubated at room temperature for 1.5 hours) with normal HEK-293 (sh-control ) and three independent sh-B7-H3 transfected HEK-293 cells.

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  WB analysis of A431 using 66481-1-Ig

  Untreated and PNGase F-treated lysates of A431 cells and PC-3 cells were subjected to SDS PAGE followed by western blot with 66481-1-Ig (B7-H3/CD276 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. PNGase F was obtained from Atagenix (cat.NO. ata808).

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  IHC staining of human tonsillitis using 66481-1-Ig

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66481-1-Ig (B7-H3/CD276 antibody) at dilution of 1:5000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human tonsillitis using 66481-1-Ig

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66481-1-Ig (B7-H3/CD276 antibody) at dilution of 1:5000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human tonsillitis using 66481-1-Ig

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66481-1-Ig (B7-H3/CD276 antibody) at dilution of 1:5000 (under 4x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human placenta using 66481-1-Ig

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 66481-1-Ig (B7-H3/CD276 antibody at dilution of 1:750 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human placenta using 66481-1-Ig

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 66481-1-Ig (B7-H3/CD276 antibody at dilution of 1:750 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human prostate cancer using 66481-1-Ig

  Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue slide using 66481-1-Ig (B7-H3/CD276 antibody at dilution of 1:300 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human prostate cancer using 66481-1-Ig

  Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue slide using 66481-1-Ig (B7-H3/CD276 antibody at dilution of 1:300 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 66481-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66481-1-Ig (B7-H3/CD276 antibody at dilution of 1:300 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 66481-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66481-1-Ig (B7-H3/CD276 antibody at dilution of 1:300 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of human placenta using 66481-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed paraffin-embedded human placenta tissue using B7-H3/CD276 antibody (66481-1-Ig, Clone: 1E7D1 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) (SA00013-1). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• Featured Product
• KD/KO Validated

B7-H3/CD276 Monoclonal Antibody for WB, IHC, IF-P, ELISA

Cat No. 66481-1-Ig

Clone No.1E7D1

Publications(12)

Reviews (1)

Host / Isotype

Mouse / IgG2a

Reactivity

human, pig

Applications

WB, IHC, IF-P, IP, CoIP, ELISA

Synonyms

CD276, B7-H3, B7H3, B7 homolog 3, B7 H3

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Tested Applications

• Positive WB detected in: HEK-293 cells, A431 cells, PC-3 cells
• Positive IHC detected in: human tonsillitis tissue, human placenta tissue, human prostate cancer tissue, human lung cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: human placenta tissue

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:6000
• Immunohistochemistry (IHC): IHC : 1:2500-1:10000
• Immunofluorescence (IF)-P: IF-P : 1:200-1:800

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 5 publications below
• IHC: See 6 publications below
• IF: See 2 publications below
• IP: See 1 publications below
• FC: See 1 publications below
• CoIP: See 2 publications below

Product Information

66481-1-Ig targets B7-H3/CD276 in WB, IHC, IF-P, IP, CoIP, ELISA applications and shows reactivity with human, pig samples.

• Tested Reactivity: human, pig
• Cited Reactivity: human
• Host / Isotype: Mouse / IgG2a
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag6006

  Product name: Recombinant human CD276 protein

  Source: e coli.-derived, PGEX-4T

  Tag: GST

  Domain: 41-181 aa of BC062581

  Sequence: LVGTDATLCCSFSPEPGFSLAQLNLIWQLTDTKQLVHSFAEGQDQGSAYANRTALFPDLLAQGNASLRLQRVRVADEGSFTCFVSIRDFGSAAVSLQVAAPYSKPSMTLEPNKDLRPGDTVTITCSSYQGYPEAEVFWQDG

• Full Name: CD276 molecule
• Calculated Molecular Weight: 57 kDa
• Observed Molecular Weight: 100 kDa
• GenBank Accession Number: BC062581
• Gene Symbol: CD276
• Gene ID (NCBI): 80381
• RRID: AB_2881847
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: Q5ZPR3
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

B7-H3 (CD276) is a type I transmembrane protein expressed on many tissues and cell types. B7-H3 is a 100-kDa glycoprotein that belongs to the B7 immunoregulatory family and participates in the regulation of T-cell-mediated immune response probably by functioning as both a T cell costimulator and coinhibitor (PMID: 25567370; 20696859). Overexpressed on a wide range of human solid cancers, B7-H3 has been implicated in cancer progression and metastasis and becomes an attractive target for cancer immunotherapy (PMID: 27208063).

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
human	WB,FC	J Immunother Cancer; Genome-scale CRISPR-Cas9 screen reveals novel regulators of B7-H3 in tumor cells.; Authors - Shasha Zhao
human	IP	J Transl Med; An N-glycoproteomic site-mapping analysis reveals glycoprotein alterations in esophageal squamous cell carcinoma.; Authors - Yingzhen Gao
human	WB,IHC	J Cancer Res Clin Oncol; B7-H3 and CD47 co-expression in gastric cancer is a predictor of poor prognosis and potential targets for future dual-targeting immunotherapy; Authors - Siji Chen
human	WB,CoIP	J Cancer; B7-H3 inhibits apoptosis of gastric cancer cell by interacting with Fibronectin.; Authors - Meiyun Sun
human	IHC	Int J Gen Med; Comprehensive Analysis Reveals Distinct Immunological and Prognostic Characteristics of CD276/B7-H3 in Pan-Cancer; Authors - Jinye Ding
human	WB,IHC,CoIP	FEBS Open Bio; Fibronectin enhances tumor metastasis through B7-H3 in clear cell renal cell carcinoma; Authors - Jinjing Xie

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Margarita (Verified Customer) (11-20-2024)

Secondary antibody: AF 488

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:400
• Cell Tissue Type: PC3 cells

B7-H3/CD276 Antibody Immunofluorescence validation (1:400 dilution) in PC3 cells (Cat no:66481-1-Ig)