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  1. Link

    cd47-antibody-epr21794-ab218810.pdf

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Immunology Cell Type Markers CD Adhesion
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD47 antibody [EPR21794] (ab218810)

  • Datasheet
  • Certificate of Compliance
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (ab218810)
  • Western blot - Anti-CD47 antibody [EPR21794] (ab218810)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (ab218810)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (ab218810)
  • Western blot - Anti-CD47 antibody [EPR21794] (ab218810)
  • Anti-CD47 antibody [EPR21794] (ab218810)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21794] to CD47
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-CD47 antibody [EPR21794]
    See all CD47 primary antibodies
  • Description

    Rabbit monoclonal [EPR21794] to CD47
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK-293T, Jurkat, U937, A549 and U-2 OS whole cell lysates. IHC-P: Human ovary carcinoma, liver and placenta 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: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR21794
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Cell Type Markers
    • CD
    • Adhesion
    • Cancer
    • Tumor immunology
    • CD markers

Associated products

  • Alternative Versions

    • Anti-CD47 antibody [EPR21794] - BSA and Azide free (ab233122)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

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

    • Human CD47 knockout HEK-293T cell line (ab266324)
  • KO cell lysates

    • Human CD47 knockout HEK-293T cell lysate (ab257220)
  • Recombinant Protein

    • Recombinant Human CD47 protein (His tag) (ab219233)
  • Related Products

    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab218810 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 47-52 kDa (predicted molecular weight: 35 kDa).
IHC-P (1)
1/2000. 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 47-52 kDa (predicted molecular weight: 35 kDa).
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Has a role in both cell adhesion by acting as an adhesion receptor for THBS1 on platelets, and in the modulation of integrins. Plays an important role in memory formation and synaptic plasticity in the hippocampus (By similarity). Receptor for SIRPA, binding to which prevents maturation of immature dendritic cells and inhibits cytokine production by mature dendritic cells. Interaction with SIRPG mediates cell-cell adhesion, enhances superantigen-dependent T-cell-mediated proliferation and costimulates T-cell activation. May play a role in membrane transport and/or integrin dependent signal transduction. May prevent premature elimination of red blood cells. May be involved in membrane permeability changes induced following virus infection.
  • Tissue specificity

    Very broadly distributed on normal adult tissues, as well as ovarian tumors, being especially abundant in some epithelia and the brain.
  • Sequence similarities

    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Cellular localization

    Cell membrane.
  • Target information above from: UniProt accession Q08722 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 961 Human
    • Omim: 601028 Human
    • SwissProt: Q08722 Human
    • Unigene: 446414 Human
    • Alternative names

      • Antigen identified by monoclonal antibody 1D8 antibody
      • Antigenic surface determinant protein OA3 antibody
      • CD 47 antibody
      • CD47 antibody
      • CD47 antigen (Rh-related antigen, integrin-associated signal transducer) antibody
      • CD47 antigen antibody
      • CD47 glycoprotein antibody
      • CD47 molecule antibody
      • CD47_HUMAN antibody
      • IAP antibody
      • Integrin Associated Protein antibody
      • Integrin associated signal transducer antibody
      • Integrin-associated protein antibody
      • Leukocyte surface antigen CD47 antibody
      • MER 6 antibody
      • MER6 antibody
      • OA 3 antibody
      • OA3 antibody
      • OTTHUMP00000041152 antibody
      • OTTHUMP00000041153 antibody
      • Protein MER6 antibody
      • Rh related antigen antibody
      • Surface antigen identified by monoclonal antibody 1D8 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (ab218810)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (ab218810)

      Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling CD47 with ab218810 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on human placenta (PMID: 7998989). Counterstained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    • Western blot - Anti-CD47 antibody [EPR21794] (ab218810)
      Western blot - Anti-CD47 antibody [EPR21794] (ab218810)
      All lanes : Anti-CD47 antibody [EPR21794] (ab218810) at 1/500 dilution

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : CD47 knockout HEK293T cell lysate
      Lane 3 : Jurkat cell lysate
      Lane 4 : HepG2 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: 35 kDa
      Observed band size: 47-52 kDa why is the actual band size different from the predicted?



      Lanes 1-4: Merged signal (red and green). Green - ab218810 observed at 47-52 kDa. Red - loading control ab8245 observed at 36 kDa.

      ab218810 Anti-CD47 antibody [EPR21794] was shown to specifically react with CD47 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266324 (knockout cell lysate ab257220) was used. Wild-type and CD47 knockout samples were subjected to SDS-PAGE. ab218810 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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-CD47 antibody [EPR21794] (ab218810)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (ab218810)

      Immunohistochemical analysis of paraffin-embedded human liver tissue labeling CD47 with ab218810 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on sinus of human liver sinusoidal endothelial cells (PMID: 7998989). Counterstained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (ab218810)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (ab218810)

      Immunohistochemical analysis of paraffin-embedded human ovary carcinoma tissue labeling CD47 with ab218810 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weakly cytoplasmic staining in human ovary carcinoma (PMID: 7998989). Counterstained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    • Western blot - Anti-CD47 antibody [EPR21794] (ab218810)
      Western blot - Anti-CD47 antibody [EPR21794] (ab218810)
      Lanes 1-4 : Anti-CD47 antibody [EPR21794] (ab218810) at 1/5000 dilution
      Lane 5 : Anti-CD47 antibody [EPR21794] (ab218810) at 1/1000 dilution

      Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
      Lane 2 : U937 (human histiocytic lymphoma cell line) whole cell lysate at 20 µg
      Lane 3 : A549 (human lung carcinoma cell line) whole cell lysate at 20 µg
      Lane 4 : U-2 OS (human bone osteosarcoma epithelial cell line) whole cell lysate at 20 µg
      Lane 5 : Human brain lysate at 10 µg

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

      Predicted band size: 35 kDa
      Observed band size: 47-52 kDa why is the actual band size different from the predicted?



       

      Exposure time: Lane 1: 26 seconds; Lane 2: 70 seconds; Lanes 3-4: 3 minutes; Lane 5: 3 seconds.

      Blocking/Dilution buffer: 5% NFDM/TBST.

      We recommend that samples are not boiled after adding loading buffer as this may cause protein aggregates. The molecular mass observed is consistent with the literature (PMID 12393467, PMID 11034562).

       

       

    • Anti-CD47 antibody [EPR21794] (ab218810)
      Anti-CD47 antibody [EPR21794] (ab218810)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

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

    References (3)

    Publishing research using ab218810? Please let us know so that we can cite the reference in this datasheet.

    ab218810 has been referenced in 3 publications.

    • Jani VP  et al. RRx-001 Increases Erythrocyte Preferential Adhesion to the Tumor Vasculature. Int J Mol Sci 22:N/A (2021). PubMed: 33946824
    • 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
    • Yang Z  et al. PD-L1 and CD47 co-expression in pulmonary sarcomatoid carcinoma: a predictor of poor prognosis and potential targets of future combined immunotherapy. J Cancer Res Clin Oncol 145:3055-3065 (2019). PubMed: 31522278

    Customer reviews and Q&As

    Show All Reviews Q&A
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    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD47 antibody [EPR21794]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Pig Tissue sections (Heat - Wild-type vs hCD47-tg)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH9.0
    Permeabilization
    No
    Specification
    Heat - Wild-type vs hCD47-tg
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 11 2022

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