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    cd276-antibody-epncir122-ab134161.pdf

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Immunology Immune Interventions Transplantation
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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-CD276 抗体 [EPNCIR122] (ab134161)

  • Datasheet
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Abpromise

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Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
  • Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (ab134161)
  • Immunocytochemistry - Anti-CD276 antibody [EPNCIR122] (ab134161)
  • Immunoprecipitation - Anti-CD276 antibody [EPNCIR122] (ab134161)
  • Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
  • Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (ab134161)
  • Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (ab134161)
  • Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
  • Immunohistochemistry (Frozen sections) - Anti-CD276 antibody [EPNCIR122] (ab134161)
  • Anti-CD276 antibody [EPNCIR122] (ab134161)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPNCIR122] to CD276
  • Suitable for: IHC-Fr, WB, IP, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

こちらの製品もご検討ください

二次抗体
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
タンパク質
Recombinant human CD276 protein (Fc Chimera Active) (ab216204)
ノックアウト
Product image
Human CD276 knockout HEK293T cell line (ab266658)

関連製品

製品の概要

  • 製品名

    Anti-CD276 antibody [EPNCIR122]
    CD276 一次抗体 製品一覧
  • 製品の詳細

    Rabbit monoclonal [EPNCIR122] to CD276
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: IHC-Fr, WB, IP, Flow Cyt, ICC/IFmore details
    適用なし: IHC-P
  • 種交差性

    交差種: Mouse, Human
  • 免疫原

    Recombinant full length protein corresponding to Human CD276. Alternate injections with Human recombinant protein and Human CD276 over-expressing CHO cells.
    Database link: Q5ZPR3
    (Peptide available as ab182198)

  • ポジティブ・コントロール

    • WB: LnCaP, CHO-K1 cells lysates transfected with human and mouse CD276. Flow Cyt: HEK293 and THP1 cells. ICC: HEK293 cells.
  • 特記事項

    This antibody was developed as part of a collaboration between Epitomics, the National Cancer Institute's Center for Cancer Research and the lab of Brad St. Croix.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • バッファー

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • 精製度

    Protein A purified
  • ポリ/モノ

    モノクローナル
  • クローン名

    EPNCIR122
  • アイソタイプ

    IgG
  • 研究分野

    • Immunology
    • Immune Interventions
    • Transplantation
    • Immunology
    • Adaptive Immunity
    • T Cells
    • CD
    • Cancer
    • Tumor biomarkers
    • Other
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • T Lymphocytic Lineage

関連製品

  • Alternative Versions

    • Anti-CD276 antibody [EPNCIR122] - Low endotoxin, Azide free (ab209895)
    • Alexa Fluor® 488 Anti-CD276 antibody [EPNCIR122] (ab218280)
    • Alexa Fluor® 594 Anti-CD276 antibody [EPNCIR122] (ab218282)
    • Alexa Fluor® 555 Anti-CD276 antibody [EPNCIR122] (ab218283)
    • Anti-CD276 antibody [EPNCIR122] - BSA and Azide free (ab256585)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CD276 knockout HEK293T cell line (ab266658)
  • Recombinant Protein

    • Recombinant human CD276 protein (Fc Chimera Active) (ab216204)
  • Related Products

    • Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)

アプリケーション

Our Abpromise guarantee covers the use of ab134161 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
IHC-Fr Use at an assay dependent concentration.
WB 1/200. Predicted molecular weight: 57 kDa.
IP 1/10 - 1/100.
Flow Cyt 1/1000 - 1/10000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use a concentration of 1 µg/ml.
  • 追加情報
    Is unsuitable for IHC-P.
  • ターゲット情報

    • 機能

      May participate in the regulation of T-cell-mediated immune response. May play a protective role in tumor cells by inhibiting natural-killer mediated cell lysis as well as a role of marker for detection of neuroblastoma cells. May be involved in the development of acute and chronic transplant rejection and in the regulation of lymphocytic activity at mucosal surfaces. Could also play a key role in providing the placenta and fetus with a suitable immunological environment throughout pregnancy. Both isoform 1 and isoform 2 appear to be redundant in their ability to modulate CD4 T-cell responses. Isoform 2 is shown to enhance the induction of cytotoxic T-cells and selectively stimulates interferon gamma production in the presence of T-cell receptor signaling.
    • 組織特異性

      Ubiquitous but not detectable in peripheral blood lymphocytes or granulocytes. Weakly expressed in resting monocytes. Expressed in dendritic cells derived from monocytes. Expressed in epithelial cells of sinonasal tissue. Expressed in extravillous trophoblast cells and Hofbauer cells of the first trimester placenta and term placenta.
    • 配列類似性

      Belongs to the immunoglobulin superfamily. BTN/MOG family.
      Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
      Contains 2 Ig-like V-type (immunoglobulin-like) domains.
    • 細胞内局在

      Membrane.
    • Target information above from: UniProt accession Q5ZPR3 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • 参照データベース

      • Entrez Gene: 80381 Human
      • Entrez Gene: 102657 Mouse
      • Omim: 605715 Human
      • SwissProt: Q5ZPR3 Human
      • SwissProt: Q8VE98 Mouse
      • Unigene: 744915 Human
      • Unigene: 77873 Human
      • Unigene: 5356 Mouse
      • 別名

        • 4Ig B7 H3 antibody
        • 4Ig-B7-H3 antibody
        • AU016588 antibody
        • B7 H3 antibody
        • B7 homolog 3 antibody
        • B7-H3 antibody
        • B7H3 antibody
        • B7RP-2 antibody
        • CD_antigen=CD276 antibody
        • CD276 antibody
        • CD276 antigen antibody
        • CD276 molecule antibody
        • CD276_HUMAN antibody
        • Costimulatory molecule antibody
        • Flags: Precursor antibody
        • PSEC0249 antibody
        • UNQ309/PRO352 antibody
        see all

      画像

      • Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Anti-CD276 antibody [EPNCIR122] (ab134161) at 1/200 dilution + CHO (hamster ovary epithelial cell) whole cell lysates transfected with human CD276 at 15 µg

        Secondary
        Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

        Predicted band size: 57 kDa
        Observed band size: 110 kDa
        why is the actual band size different from the predicted?


        Exposure time: 3 minutes


        Blocking/dilution buffer: 5% NFDM/TBST

        Performed using purified antibody.

      • Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (ab134161)

        Flow cytometry overlay histogram showing wild-type HEK293 (green line) and CD276 knockout HEK293 (ab266658) cells stained with ab134161 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab134161) (1x106 in 100μl at 0.2 μg/ml) for 30 min at 4°C.

        The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150081) was used at 1/2000 for 30 min at 4°C.

        Isotype control antibody was Rabbit IgG (monoclonal) (ab172730) used at the same concentration and conditions as the primary antibody (wild-type HEK293 - black line CD276 HEK293 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

        Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

      • Immunocytochemistry - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Immunocytochemistry - Anti-CD276 antibody [EPNCIR122] (ab134161)
        ab134161 staining CD276 in wild-type HEK293 cells (top panel) and CD276 knockout HEK293 cells (ab266658) (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab134161 at 1μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
        Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
      • Immunoprecipitation - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Immunoprecipitation - Anti-CD276 antibody [EPNCIR122] (ab134161)

        CD276 was immunoprecipitated from 10 ug of HEK 293 (human embryonic kidney epithelial cell) whole cell lysate with ab134161 at 1/40 diltuion. Western blot was permformed from the immunoprecipitate using ab 134161 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/500 dilution.

        Lane 1: HEK 293 whole cell lysate 10 ug (Input).

        Lane 2: ab134161 IP in HEK 293 whole cell lysate.

        Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab134161 in HEK 293 whole cell lysate.

        Blocking/dilution buffer: 5% NFDM/TBST

        Performed using purified antibody.

      • Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Anti-CD276 antibody [EPNCIR122] (ab134161) at 1/1000 dilution + LNCap (human prostate carcinoma epithelial cell) whole cell lysate at 15 µg

        Secondary
        Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

        Predicted band size: 57 kDa
        Observed band size: 110 kDa why is the actual band size different from the predicted?


        Exposure time: 3 minutes


        Blocking/dilution buffer: 5% NFDM/TBST

        Performed using purified antibody.

      • Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (ab134161)

        Overlay histogram showing THP-1 (human monocytic leukemia monocyte) cells stained with ab134161 (red line). The cells were fixed with 4% paraformaldehyde and then permeabilized with 90% methanol. The cells were incubated with the antibody (ab134161) at 1/80 dilution. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution. Isotype control antibody (black line) was rabbit monoclonal IgG (ab172730) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

        Performed using purified antibody.

      • Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Flow Cytometry - Anti-CD276 antibody [EPNCIR122] (ab134161)

        Overlay histogram showing THP1 cells stained with ab134161 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% human serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab134161, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

      • Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Western blot - Anti-CD276 antibody [EPNCIR122] (ab134161)
        All lanes : Anti-CD276 antibody [EPNCIR122] (ab134161) at 0.264 µg/ml

        Lane 1 : CHO-K1 cell lysates, transfected with Human CD276
        Lane 2 : CHO-K1 cell lysates, transfected with Mouse CD276.

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

        Predicted band size: 57 kDa


        Exposure time: 3 minutes


        Blocking/dilution buffer: 5% NFDM/TBST

      • Immunohistochemistry (Frozen sections) - Anti-CD276 antibody [EPNCIR122] (ab134161)
        Immunohistochemistry (Frozen sections) - Anti-CD276 antibody [EPNCIR122] (ab134161)Image from Seaman S et al. Cancer Cell, 31, 501-515. Fig2.D doi:10.1016/j.ccell.2017.03.005. with permission from Elsevier

        Immunohistochemistry of mouse MC38 colon liver metastasis. Staining CD276 with ab134161 (green). Normal tissue (N)/tumor tissue (T) margins are indicated by a white dash.

         

      • Anti-CD276 antibody [EPNCIR122] (ab134161)
        Anti-CD276 antibody [EPNCIR122] (ab134161)

      プロトコール

      • Flow cytometry protocols
      • Immunoprecipitation protocols
      • Immunohistochemistry protocols
      • Western blot protocols

      Click here to view the general protocols

      データシートおよび資料

      • Datasheet
      • SDS
    • 参考文献 (10)

      ab134161 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

      ab134161 は 10 報の論文で使用されています。

      • Bachawal S  et al. Evaluation of ductal carcinoma in situ grade via triple-modal molecular imaging of B7-H3 expression. NPJ Breast Cancer 6:14 (2020). PubMed: 32377564
      • Du H  et al. Antitumor Responses in the Absence of Toxicity in Solid Tumors by Targeting B7-H3 via Chimeric Antigen Receptor T Cells. Cancer Cell 35:221-237.e8 (2019). PubMed: 30753824
      • Xu ZL  et al. B7-H3 promotes malignant progression of muscle-invasive bladder cancer. Oncol Rep 40:2722-2733 (2018). PubMed: 30132557
      • Wilson KE  et al. Intraoperative Resection Guidance with Photoacoustic and Fluorescence Molecular Imaging Using an Anti-B7-H3 Antibody-Indocyanine Green Dual Contrast Agent. Clin Cancer Res 24:3572-3582 (2018). PubMed: 29712688
      • Xu Z  et al. High expression of B7-H3 and CD163 in cancer tissues indicates malignant clinicopathological status and poor prognosis of patients with urothelial cell carcinoma of the bladder. Oncol Lett 15:6519-6526 (2018). PubMed: 29725402
      • Seaman S  et al. Eradication of Tumors through Simultaneous Ablation of CD276/B7-H3-Positive Tumor Cells and Tumor Vasculature. Cancer Cell 31:501-515.e8 (2017). WB, IP, IHC-Fr, Flow Cyt ; Mouse, Human . PubMed: 28399408
      • Wilson KE  et al. Spectroscopic Photoacoustic Molecular Imaging of Breast Cancer using a B7-H3-targeted ICG Contrast Agent. Theranostics 7:1463-1476 (2017). PubMed: 28529630
      • Zhang W  et al. B7-H3 silencing inhibits tumor progression of mantle cell lymphoma and enhances chemosensitivity. Int J Oncol 46:2562-72 (2015). WB . PubMed: 25872657
      • Zhang W  et al. B7-H3 silencing by RNAi inhibits tumor progression and enhances chemosensitivity in U937 cells. Onco Targets Ther 8:1721-33 (2015). WB ; Human . PubMed: 26203263
      • Hasegawa K  et al. Chromosomal gain promotes formation of a steep RanGTP gradient that drives mitosis in aneuploid cells. J Cell Biol 200:151-61 (2013). WB, ICC/IF ; Human . PubMed: 23319601

      レビューと Q&A

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      There are currently no Customer reviews or Questions for ab134161.
      Please use the links above to contact us or submit feedback about this product.

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com

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