Human CD47 Antibody

  (2 citations)
(1 Review)
    
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Citations (2)
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  • Species Reactivity
    Human
  • Specificity
    Detects human CD47 in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 5% cross-reactivity with recombinant mouse CD47 is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human CD47
    Gln19-Pro139
    Accession # Q08722
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Product Datasheets

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Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Flow Cytometry
    2.5 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
    CD47 is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in CD47 knockout HEK293T cell line.
  • Neutralization
    Measured by its ability to neutralize SIRP alpha /CD172a-mediated adhesion of human erythrocytes.  The adhesion of human erythrocytes to immobilized Recombinant Human SIRP alpha /CD172a/Fc Chimera (2 µg/mL, 100 µL/well) was maximally inhibited (70-100%) by 2 µg/mL of the antibody.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human CD47 by Western Blot. Western blot shows lysates of U937 human histiocytic lymphoma cell line and human placenta tissue, not heated to minimize aggregation. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human CD47 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4670) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for CD47 at approximately 45-70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of CD47 in Human Lymphocytes by Flow Cytometry. Human whole blood lymphocytes were stained with Sheep Anti-Human CD47 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF4670, filled histogram) or control antibody (Catalog # 5-001-A, open histogram), followed by Northern­Lights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # NL010).
Immunohistochemistry
CD47 in Human Placenta. CD47 was detected in immersion fixed paraffin-embedded sections of human placenta using 5 µg/mL Sheep Anti-Human CD47 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4670) overnight at 4 °C. Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Cell Adhesion Mediated by SIRP alpha /CD172a and Neutral­ization by Human CD47 Antibody. Recombinant Human SIRP alpha /CD172a (Catalog # 4546-SA), immobilized onto a microplate, supports the adhesion of the human erythrocytes in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Human SIRP-alpha (2 µg/mL) is neutralized (green line) by increasing concentrations of Sheep Anti-Human CD47 Poly­clonal Antibody (Catalog # AF4670). The adhesion was maximally inhibited (70-100%) by 2 µg/mL of the antibody.
Knockout Validated
Western Blot Shows Human CD47 Specificity by Using Knockout Cell Line. Western blot shows lysates of HEK293T human embryonic kidney parental cell line and CD47 knockout HEK293T cell line (KO). PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human CD47 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4670) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for CD47 at approximately 50 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockoutHEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
    Reconstitution Buffer Available
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: CD47

CD47 (also integrin-associated protein/IAP and OA3) is a variably glycosylated, 40‑60 kDa atypical member of the Ig-Superfamily. It is expressed on almost all cell types, including erythrocytes. CD47 binds to TSP-1 and SIRP alpha, and forms a membrane complex with CD36 and  alpha v beta 3. Mature human CD47 is a 305 amino acid (aa), five-transmembrane glycoprotein. It contains a 123 aa extracellular region (aa 19‑141) that is characterized by the presence of a V-type Ig-like domain (aa 19‑127), and a 34 aa C-terminal cytoplasmic tail that interacts with Gi alpha subunits. Three splice variants occur over aa 293‑323. Over aa 19‑139, human CD47 shares 61%, 71% and 66% aa identity with mouse, porcine and canine CD47, respectively.

  • Entrez Gene IDs:
    961 (Human); 16423 (Mouse); 102139846 (Cynomolgus Monkey)
  • Alternate Names:
    antigen identified by monoclonal 1D8; Antigenic surface determinant protein OA3; CD47 antigen (Rh-related antigen, integrin-associated signal transducer); CD47 antigen; CD47 glycoprotein; CD47 molecule; CD47; IAP; IAPintegrin associated protein; Integrin-associated protein; leukocyte surface antigen CD47; MER6integrin-associated signal transducer; OA3; Protein MER6; Rh-related antigen
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Species
Applications
Sample Type
  1. Relationship between tumor-associated macrophage subsets and CD47 expression in squamous cell carcinoma of the head and neck in the tumor microenvironment
    Authors: Koichi Sakakura
    Lab Invest, 2016;0(0):.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  2. Identification of a population of blood circulating tumor cells from breast cancer patients that initiates metastasis in a xenograft assay.
    Authors: Baccelli I, Schneeweiss A, Riethdorf S, Stenzinger A, Schillert A, Vogel V, Klein C, Saini M, Bauerle T, Wallwiener M, Holland-Letz T, Hofner T, Sprick M, Scharpff M, Marme F, Sinn H, Pantel K, Weichert W, Trumpp A
    Nat Biotechnol, 2013;31(6):539-44.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded

FAQs

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Cell and Tissue Staining Kits
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Anti-Sheep HRP-DAB Cell & Tissue Staining Kit

CTS019 2
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Immunohistochemistry Reagents
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Mounting Medium

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Antigen Retrieval Reagent Sampler (50 mL each)

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VisUCyte Antigen Retrieval Reagent-Acidic

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VisUCyte Antigen Retrieval Reagent-Universal

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Isotype Controls
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Normal Sheep IgG Control

Ctrl 5-001-A 11
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Reconstitution Buffers
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Reconstitution Buffer 1 (PBS)

RB01
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Secondary Antibodies
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Sheep IgG HRP-conjugated Antibody

WB , Simple Western HAF016 18  
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Donkey Anti-Sheep IgG NorthernLights™ NL557-conjugated Antibody

Flow , IHC , ICC NL010 3
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Donkey Anti-Sheep IgG NorthernLights™ NL637-conjugated Antibody

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Donkey Anti-Sheep IgG NorthernLights™ NL493-conjugated Antibody

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Sheep IgG (H+L) PE-conjugated Antibody

Flow F0126  
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Sheep IgG (H+L) APC-conjugated Antibody

Flow F0127  
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Donkey Anti-Sheep IgG Biotinylated Antibody

WB BAF016 2
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Sheep IgG VisUCyte HRP Polymer Antibody

IHC VC006  
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Donkey Anti-Sheep IgG (H+L) PerCP-conjugated Antibody

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Average Rating: 5 (Based on 1 review)

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We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Western Blot.

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Images Ratings Applications Species Reviewed By Date Details
Western Blot Human CD47 Antibody AF4670 Array AF4670
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Excellent
 WB Human Anonymous 07/15/2018
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Western Blot Human CD47 Antibody AF4670
Western Blot: Human CD47 Antibody [AF4670].

Summary

Application Western Blot
Sample Tested HepG2 human hepatocellular carcinoma cell line
Species Human

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