抗B7-H3抗体(Anti-B7-H3, Human, Goat-Poly antibody)

• 価格
• Product Details
• Applications and Data
• Related Product & Information
• Citations

価格

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Product Details

Species Reactivity	Human
Label	Unconjugated
Immunogen	Mouse myeloma cell line NS0-derived recombinant human B7‑H3Leu29-Pro245Accession # NP_079516
Source	Polyclonal Goat IgG
Purification	Antigen Affinity-purified
Specificity	Detects human B7-H3 in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross-reactivity with recombinant human (rh) B7‑H2 is observed, and less than 1% cross-reactivity with rhB7-1, rhB7-2, rhB7-H1, and rhPD-L2 is observed.

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Applications and Data

	Recommended Concentration	Sample
Western Blot	1 µg/mL	See below
Simple Western	10 µg/mL	See below
Flow Cytometry	0.25 µg/106 cells	See below
Immunohistochemistry	5-15 µg/mL	See below
CyTOF-ready	Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Knockout Validated	B7‑H3 is specifically detected in U2OS human osteosarcoma parental cell line but is not detectable in B7‑H3 knockout U2OS cell line.

Western Blot
	Detection of Human B7‑H3 by Western Blot. 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.
Flow Cytometry
	Detection of B7‑H3 in PC‑3 Human Cell Line by Flow Cytometry. 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).
Immunohistochemistry
	B7-H3 in Human Melanoma. 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.
Simple Western
	Detection of Human B7‑H3 by Simple WesternTM. 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 approximately120-160 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.
Knockout Validated
	Western Blot Shows Human B7‑H3 Specificity by Using Knockout Cell Line. 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.

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Related Product & Information

Background

B7-H3

background_content

Background: B7-H3 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).

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Citations

1. B7-H3 Promotes the Migration and Invasion of Human Bladder Cancer Cells via the PI3K/Akt/STAT3 Signaling Pathway
   Authors: Y Li, G Guo, J Song, Z Cai, J Yang, Z Chen, Y Wang, Y Huang, Q Gao
   J Cancer, 2017;8(5):816-824.
   Species: Human
   Sample Type: Whole Tissue
   Application: IHC Paraffin-embedded


2. Spectroscopic Photoacoustic Molecular Imaging of Breast Cancer using a B7-H3-targeted ICG Contrast Agent
   Authors: KE Wilson, SV Bachawal, L Abou-Elkac, K Jensen, S Machtaler, L Tian, JK Willmann
   Theranostics, 2017;7(6):1463-1476.
   Species: Human
   Sample Type: Whole Tissue
   Application: IHC


3. High constitutive B7-H3 expression on human keratinocytes supports T cell immunity
   Authors: D Quandt, E Fiedler, A Müller, WC Marsch, B Seliger
   J. Dermatol. Sci., 2017;0(0):.
   Species: Human
   Sample Type: Whole Tissue
   Application: IHC


4. Preferential Induction of the T Cell Auxiliary Signaling Molecule B7-H3 on Synovial Monocytes in Rheumatoid Arthritis
   Authors: BR Yoon, YH Chung, SJ Yoo, K Kawara, J Kim, IS Yoo, CG Park, SW Kang, WW Lee
   J. Biol. Chem, 2016;291(8):4048-57.
   Species: Human
   Sample Type: Cell Lysates
   Application: WB


5. Breast Cancer Detection by B7-H3-Targeted Ultrasound Molecular Imaging.
   Authors: Bachawal S, Jensen K, Wilson K, Tian L, Lutz A, Willmann J
   Cancer Res, 2015;75(12):2501-9.
   Species: Human
   Sample Type: Whole Tissue
   Application: IHC Paraffin-embedded


6. Origination of new immunological functions in the costimulatory molecule B7-H3: the role of exon duplication in evolution of the immune system.
   Authors: Sun J, Fu F, Gu W, Yan R, Zhang G, Shen Z, Zhou Y, Wang H, Shen B, Zhang X
   PLoS ONE, 2011;6(9):e24751.
   Species: Human
   Sample Type: Cell Lysates
   Application: WB


7. Tumor cell and tumor vasculature expression of B7-H3 predict survival in clear cell renal cell carcinoma.
   Authors: Crispen PL, Sheinin Y, Roth TJ, Lohse CM, Kuntz SM, Frigola X, Thompson RH, Boorjian SA, Dong H, Leibovich BC, Blute ML, Kwon ED
   Clin. Cancer Res., 2008;14(16):5150-7.
   Species: Human
   Sample Type: Whole Tissue
   Application: IHC Paraffin-embedded


8. Anterior pituitary progenitor cells express costimulatory molecule 4Ig-B7-H3.
   Authors: Nagai Y, Aso H, Ogasawara H, Tanaka S, Taketa Y, Watanabe K, Ohwada S, Rose MT, Kitazawa H, Yamaguchi T
   J. Immunol., 2008;181(9):6073-81.
   Species: Bovine
   Sample Type: Cell Lysates
   Application: WB


9. Interactions of T cells with fibroblast-like synoviocytes: role of the B7 family costimulatory ligand B7-H3.
   Authors: Tran CN, Thacker SG, Louie DM, Oliver J, White PT, Endres JL, Urquhart AG, Chung KC, Fox DA
   J. Immunol., 2008;180(5):2989-98.
   Species: Human
   Sample Type: Whole Cells
   Application: ICC


10. 12-O-tetradecanoyl phorbol 13-acetate induces the expression of B7-DC, -H1, -H2, and -H3 in K562 cells.
    Authors: Jang BC, Park YK, Choi IH, Kim SP, Hwang JB, Baek WK, Suh MH, Mun KC, Suh SI
    Int. J. Oncol., 2007;31(6):1439-47.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow

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