Overview

  • Product nameAnti-CD47 antibody [B6H12.2]See all CD47 primary antibodies ...
  • Description
    Mouse monoclonal [B6H12.2] to CD47
  • Tested applicationsFlow Cyt, Functional Studies, ICC, IP, ICC/IF, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Intact CD47 purified from placenta.

  • EpitopeIg domain.
  • Positive control
    • Tonsil.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer10mM PBS, pH7.4, 0.2%BSA, 0.09% sodium azide
  • Concentration information loading...
  • PurityProtein G purified
  • Clonality Monoclonal
  • Clone numberB6H12.2
  • IsotypeIgG1
  • Light chain typekappa
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab3283 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
Flow Cyt Use 1µg for 106 cells.
Functional Studies Use at an assay dependent dilution. This antibody blocks the binding of SIRP alpha (Yoshida, et al.; Latour et al.) and inhibits in assays where CD47-integrin association is required.
ICC Use at an assay dependent dilution.
IP Use at an assay dependent dilution. (native, with protein G): use at a concentration of 2µg / mg protein lysate.
ICC/IF Use at an assay dependent dilution. PubMed: 20923773
IHC-Fr Use at an assay dependent concentration. PubMed: 22451919
WB Use a concentration of 1 µg/ml. Detects a band of approximately 49 kDa (predicted molecular weight: 35 kDa).

Target

  • FunctionHas 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 specificityVery broadly distributed on normal adult tissues, as well as ovarian tumors, being especially abundant in some epithelia and the brain.
  • Sequence similaritiesContains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Cellular localizationCell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Antigenic surface determinant protein OA3 antibody
    • CD 47 antibody
    • CD47 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
    • 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
    see all

Anti-CD47 antibody [B6H12.2] images

  • ab3283 staining CD47 in HUVEC cells by Immunocytochemistry/ Immunofluorescence.
    The cells were fixed with 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.25% Triton X-100 for 5 minutes and blocked with 1% BSA in 0.1% Tween 20 and PBS for 1 hour at room temperature. The cells were incubated in a mixture of two primary antibodies ab3283 and a rabbit anti-human VEGFR2 overnight at 4 °C. The cells were then incubated with a mixture of two secondary antibodies (Alexa Fluor 488 anti-rabbit and Alexa Fluor 594 anti-mouse) in 1% BSA for 1 hour at room temperature in the dark. Counterstained with DAPI.
  • ab3283 at 1µg/ml staining CD47 from human OVCA3 cells by ICC. The cells were methanol fixed, blocked with 3% serum and incubated with the primary antibody for 30 minutes. A biotinylated horse anti-mouse IgG was used as the secondary antibody.

    See Abreview

  • Anti-CD47 antibody [B6H12.2] (ab3283) at 1 µg/ml + Brain (Human) Tissue Lysate - adult normal tissue (ab29466) at 10 µg

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 35 kDa
    Observed band size : 49 kDa (why is the actual band size different from the predicted?)


    Exposure time : 4 minutes
  • Overlay histogram showing peripheral blood lymphocytes stained with ab3283 (red line). The cells were incubated with the antibody (ab3283, 1µg/1x106 cells) for 30 min at 4ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H&L) (ab96879) at 1/200 dilution for 30 min at 4ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed gating on peripheral blood lymphocytes.

References for Anti-CD47 antibody [B6H12.2] (ab3283)

This product has been referenced in:
  • Edris B  et al. Antibody therapy targeting the CD47 protein is effective in a model of aggressive metastatic leiomyosarcoma. Proc Natl Acad Sci U S A 109:6656-61 (2012). IHC-Fr . Read more (PubMed: 22451919) »
  • Willingham SB  et al. The CD47-signal regulatory protein alpha (SIRPa) interaction is a therapeutic target for human solid tumors. Proc Natl Acad Sci U S A 109:6662-7 (2012). IHC-Fr ; Human . Read more (PubMed: 22451913) »

See all 12 Publications for this product

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Non-reduced Non-Denaturing (Native)
Sample Cow Serum (FBS)
Specification FBS
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jun 11 2014

These products are all mouse monoclonal against CD-47. It appears that ab3283 and ab140325 are the same clone. Clone B6H12.2 is first described in: The Role of CD47 in Neutrophil Transmigration increased rate of migration correlates with increased ...

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Thank you for contacting Abcam.

As we discussed on the phone, the immunogen sequences for ab53444, ab9089, and ab3283 are unknown due to all 3 immunogens arising from tissue and cell preparations.

For CD47 (ab9089 and ab3283) the huma...

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The protocol for incubating the antibody with the protein is the same as incubating the antibody with peptide. Ideally, you want to inucbate the antibody with a molar excess of 50-fold more protein per antibody binding site to increase the probability ...

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Thank you for contacting us.

The antibody ab3283 is raised against full-length CD47 and the epitope the antibody recognizes has not been mapped, so there is no blocking peptide available. That is also the case for several other CD47 antibodi...

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DISCOUNT CODE: ***
Expiration date: November 30th, 2012

I am very pleased to hear you would like to accept our offer and test ab3283 in IHC-P. This code will give you: 1 freeprimary antibody before the expiration date. To redeem this off...

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Thank you for your email.

Please check the following link carefully; http://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount

You have to purchase ab3283 and after submitting Abreview the...

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Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guaran...

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Thank you for contacting us.

This antibody recognizes the Ig domain. According to the information provided by Uniprot for Q08722 the Ig domain is located within aa 19-127, which is in the extracellular domain.

I hope this informatio...

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Thank you for contacting Abcam.

The products ab3283 and ab70328 have not been tested for cross reactivity withcynomolgus monkeys. I was unable to find the associated cynomolgis proteins to the human proteins that many of the CD47 and HLAC1 a...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"