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    products/primary-antibodies/cd276-antibody-epr20115-ab219648.pdf

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Immunology Immune Interventions Transplantation
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD276 antibody [EPR20115] (ab219648)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (1) Submit a question References (1)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)
  • Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)
  • Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
  • Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
  • Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)
  • Anti-CD276 antibody [EPR20115] (ab219648)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20115] to CD276
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Primary
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Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Protein
Recombinant human CD276 protein (Fc Chimera Active) (ab216204)

View more associated products

Overview

  • Product name

    Anti-CD276 antibody [EPR20115]
    See all CD276 primary antibodies
  • Description

    Rabbit monoclonal [EPR20115] to CD276
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: LNCaP, U-2OS, PC-3. 293T and MDA-MB-435S whole cell lysates; Human lung, skin and placenta lysates. IHC-P: Human tonsil, placenta, hepatocellular carcinoma, lung carcinoma and breast cancer tissues.
  • General notes

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20115
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Alternative Versions

    • Anti-CD276 antibody [EPR20115] - BSA and Azide free (ab227577)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

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

    • Human CD276 knockout HEK-293T cell line (ab266658)
  • KO cell lysates

    • Human CD276 knockout HEK-293T cell lysate (ab257097)
  • Recombinant Protein

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

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab219648 in the following tested applications.

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

Application Abreviews Notes
WB
1/1000. Detects a band of approximately 90-110 kDa (predicted molecular weight: 57 kDa).
IHC-P (1)
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
WB
1/1000. Detects a band of approximately 90-110 kDa (predicted molecular weight: 57 kDa).
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    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.
  • Tissue specificity

    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.
  • Sequence similarities

    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.
  • Cellular localization

    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
  • Database links

    • Entrez Gene: 80381 Human
    • Omim: 605715 Human
    • SwissProt: Q5ZPR3 Human
    • Unigene: 77873 Human
    • Alternative names

      • 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

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)

      Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Membranous staining on dendritic cells of human tonsil is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
      Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
      All lanes : Anti-CD276 antibody [EPR20115] (ab219648) at 1/1000 dilution

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : CD276 knockout HEK293T cell lysate
      Lane 3 : LNCaP cell lysate
      Lane 4 : Raji cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 57 kDa
      Observed band size: 90-110 kDa why is the actual band size different from the predicted?



      Lanes 1-4: Merged signal (red and green). Green - ab219648 observed at 90-110 kDa. Red - loading control ab8245 observed at 36 kDa.

      ab219648 Anti-CD276 antibody [EPR20115] was shown to specifically react with CD276 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266658 (knockout cell lysate ab257097) was used. Wild-type and CD276 knockout samples were subjected to SDS-PAGE. ab219648 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)

      Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Cytoplasmic and membranous staining on vascular endothelial cells and trophoblast cells of human placenta is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
      Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
      All lanes : Anti-CD276 antibody [EPR20115] (ab219648) at 1/1000 dilution

      Lane 1 : LNCaP (Human prostate cancer cell line) whole cell lysate
      Lane 2 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 57 kDa
      Observed band size: 90-110 kDa why is the actual band size different from the predicted?


      Exposure time: 3 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      Raji cells were used as a negative control for CD276 (B7-H3) (PMID: 21344157).

       Negative control: lane 2 Raji cell lysate

    • Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
      Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
      All lanes : Anti-CD276 antibody [EPR20115] (ab219648) at 1/2000 dilution

      Lane 1 : U-2OS (Human bone osteosarcoma epithelial cell line) whole cell lysate
      Lane 2 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate
      Lane 3 : 293T (Human epithelial cell line from embryonic kidney) whole cell lysate
      Lane 4 : MDA-MB-435S (Human ductal carcinoma cell line) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 57 kDa
      Observed band size: 90-110 kDa why is the actual band size different from the predicted?


      Exposure time: 3 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      CD276 (B7-H3) has a molecular weight of about 90–100 kDa, depending on the glycosylation levels. [PMID:22473715].

    • Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
      Western blot - Anti-CD276 antibody [EPR20115] (ab219648)
      All lanes : Anti-CD276 antibody [EPR20115] (ab219648) at 1/1000 dilution

      Lane 1 : Human lung lysate
      Lane 2 : Human skin lysate
      Lane 3 : Human placenta lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

      Predicted band size: 57 kDa
      Observed band size: 90-110 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1/2: 30 seconds; Lane 3: 10 seconds.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)

      Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Negative tissue: no staining on human skeletal muscle. [PMID: 18975328].

      Counter stained with Hematoxylin.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)

      Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Membranous staining on human hepatocellular carcinoma is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)

      Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Membranous staining on human lung carcinoma is observed [PMID: 25609202].

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD276 antibody [EPR20115] (ab219648)

      Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling CD276 with ab219648 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Membranous and cytoplasmic staining on human breast cancer tumor cells and stromal cells is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Anti-CD276 antibody [EPR20115] (ab219648)
      Anti-CD276 antibody [EPR20115] (ab219648)

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (1)

    Publishing research using ab219648? Please let us know so that we can cite the reference in this datasheet.

    ab219648 has been referenced in 1 publication.

    • Zhang L  et al. Immune Checkpoint-Associated Locations of Diffuse Gliomas Comparing Pediatric With Adult Patients Based on Voxel-Wise Analysis. Front Immunol 12:582594 (2021). PubMed: 33815356

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD276 antibody [EPR20115]

    Good
    Abreviews
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Skin)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH 6.0
    Permeabilization
    No
    Specification
    Skin
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Oct 22 2018

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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