Human CD47 Antibody

Catalog # Availability Size / Price Qty
AF4670
AF4670-SP
Detection of Human CD47 by Western Blot.
10 Images
Product Details
Citations (14)
FAQs
Supplemental Products
Reviews (3)

Human CD47 Antibody Summary

Species Reactivity
Human
Specificity
Detects human CD47 in direct ELISAs and Western blots. In direct ELISAs, 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 either lyophilized or 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

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.

Scientific Data

Western Blot Detection of Human CD47 antibody by Western Blot. View Larger

Detection of Human CD47 by Western Blot. fWestern 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.

Flow Cytometry Detection of CD47 antibody in Human Lymphocytes antibody by Flow Cytometry. View Larger

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 Northern-Lights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # NL010).

Immunohistochemistry CD47 antibody in Human Placenta by Immunohistochemistry (IHC-P). View Larger

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.

Neutralization Cell Adhesion Mediated by SIRP alpha /CD172a and Neutral-ization by Human CD47 Antibody. View Larger

Cell Adhesion Mediated by SIRP alpha /CD172a and Neutral-ization by Human CD47 Antibody. Recombinant Human SIRPa/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-a (2 µg/mL) is neutralized (green line) by increasing concentrations of Sheep Anti-Human CD47 Poly-clonal Antibody (Catalog # AF4670). The adhesion was maximally inhibited (70-100%) by 2 µg/mL of the antibody.

Knockout Validated Western Blot Shows Human CD47 Antibody Specificity by Using Knockout Cell Line. View Larger

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 knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western Blot Detection of Human CD47 by Western Blot View Larger

Detection of Human CD47 by Western Blot Increased CD47 expression in aging and replicative senescence. (A) Quantification of relative CD47 mRNA levels by qPCR. Expression levels of senescent cells were normalized to the respective proliferating control (white bar). Data are representative of three independent experiments. All values are means ± SEM. **P < 0.005, ***P < 0.0005, ****P < 0.0001. Statistically significant differences were determined by unpaired Student’s t test. (B) Representative immunofluorescence images from indicated cell types stained for CD47 (green) and Hoechst (blue). Scale bar: 20 µm, except for 3T3 cells: 50 µm. (C) Whole-cell lysates from indicated cell types were isolated and analyzed by Western blotting for the indicated proteins. Senescence was induced by chemical treatment (palbociclib, except for A549: etopsoside). Shown are representative blots of three independent experiments. GAPDH images are derived from the same blot as Fig. S3 (for Panc1 cells the same GAPDH loading control is additionally used in Fig. 8 G). (D) Representative immunofluorescence images of proliferating or senescent small epithelial cells (SAEC) stained for CD47 (green) and Hoechst (blue). scale bars indicate 20 µm. n = 2–3. (E) Western blot analysis of CD47 expression in SAEC cells. Senescence was induced by chemical treatment (palbociclib). GAPDH was used as loading control. n = 2. GAPDH images are derived from the same blot as Fig. S3. (F) Representative immunofluorescence images of primary lung fibroblast (proliferative or replicative senescent by serially passaging; normal healthy [NHLF] or IPF-derived [IPF]) showing CD47 (green) and Hoechst (blue) staining. scale bars indicate 20 µm. n = 2. (G) Whole-cell lysates from primary NHLF or IPF lung fibroblasts were isolated and analyzed by Western blotting for the indicated proteins. Senescence was induced by replicative passing. Shown are representative blots of three to four independent experiments. Source data are available for this figure: SourceData F7. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/36459066), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunocytochemistry/ Immunofluorescence Detection of Human CD47 by Immunocytochemistry/Immunofluorescence View Larger

Detection of Human CD47 by Immunocytochemistry/Immunofluorescence Adhesion of EpCAMpos/neg cells on multi-marker arrays.2x105 EpCAMpos (cell pool of MCF7, SKBR3, HCC1500, ZR-75-1, TMX2-28) (A) and EpCAMlow/neg cells (MDA-MB-231) (B) were either stained with 1 μM MitoTracker Green FM (A) or MitoTracker Orange CM (B) and incubated for cell adhesion experiments on NEXTERION slides AL, coated with different antibodies and ECM molecules, alone and in combination (0.1 mg/ml each). The labeling above the single spots indicates respective capture molecules (Iso = isotype control, ms = mouse, Lam = laminin, Col = collagen, HA = hyaluronic acid, rt = rat, T = Trop2, EpC = EpCAM, 49f = CD49f, rb = rabbit). (C) Overlay image of (A) and (B); scale bars (white) = 500 μm, 20x magnification. (D) Array layout (5x5 mm) with 36 spots (spot diameter = 500 μm; pitch = 800 μm) printed on NEXTERION slides AL. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26695635), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Human CD47 by Western Blot View Larger

Detection of Human CD47 by Western Blot Increased CD47 expression in aging and replicative senescence. (A) Quantification of relative CD47 mRNA levels by qPCR. Expression levels of senescent cells were normalized to the respective proliferating control (white bar). Data are representative of three independent experiments. All values are means ± SEM. **P < 0.005, ***P < 0.0005, ****P < 0.0001. Statistically significant differences were determined by unpaired Student’s t test. (B) Representative immunofluorescence images from indicated cell types stained for CD47 (green) and Hoechst (blue). Scale bar: 20 µm, except for 3T3 cells: 50 µm. (C) Whole-cell lysates from indicated cell types were isolated and analyzed by Western blotting for the indicated proteins. Senescence was induced by chemical treatment (palbociclib, except for A549: etopsoside). Shown are representative blots of three independent experiments. GAPDH images are derived from the same blot as Fig. S3 (for Panc1 cells the same GAPDH loading control is additionally used in Fig. 8 G). (D) Representative immunofluorescence images of proliferating or senescent small epithelial cells (SAEC) stained for CD47 (green) and Hoechst (blue). scale bars indicate 20 µm. n = 2–3. (E) Western blot analysis of CD47 expression in SAEC cells. Senescence was induced by chemical treatment (palbociclib). GAPDH was used as loading control. n = 2. GAPDH images are derived from the same blot as Fig. S3. (F) Representative immunofluorescence images of primary lung fibroblast (proliferative or replicative senescent by serially passaging; normal healthy [NHLF] or IPF-derived [IPF]) showing CD47 (green) and Hoechst (blue) staining. scale bars indicate 20 µm. n = 2. (G) Whole-cell lysates from primary NHLF or IPF lung fibroblasts were isolated and analyzed by Western blotting for the indicated proteins. Senescence was induced by replicative passing. Shown are representative blots of three to four independent experiments. Source data are available for this figure: SourceData F7. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/36459066), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunocytochemistry/ Immunofluorescence Detection of Human CD47 by Immunocytochemistry/Immunofluorescence View Larger

Detection of Human CD47 by Immunocytochemistry/Immunofluorescence Adhesion of EpCAMpos/neg cells on multi-marker arrays.2x105 EpCAMpos (cell pool of MCF7, SKBR3, HCC1500, ZR-75-1, TMX2-28) (A) and EpCAMlow/neg cells (MDA-MB-231) (B) were either stained with 1 μM MitoTracker Green FM (A) or MitoTracker Orange CM (B) and incubated for cell adhesion experiments on NEXTERION slides AL, coated with different antibodies and ECM molecules, alone and in combination (0.1 mg/ml each). The labeling above the single spots indicates respective capture molecules (Iso = isotype control, ms = mouse, Lam = laminin, Col = collagen, HA = hyaluronic acid, rt = rat, T = Trop2, EpC = EpCAM, 49f = CD49f, rb = rabbit). (C) Overlay image of (A) and (B); scale bars (white) = 500 μm, 20x magnification. (D) Array layout (5x5 mm) with 36 spots (spot diameter = 500 μm; pitch = 800 μm) printed on NEXTERION slides AL. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26695635), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunocytochemistry/ Immunofluorescence Detection of Human CD47 by Immunocytochemistry/Immunofluorescence View Larger

Detection of Human CD47 by Immunocytochemistry/Immunofluorescence Adhesion of EpCAMpos/neg cells on multi-marker arrays.2x105 EpCAMpos (cell pool of MCF7, SKBR3, HCC1500, ZR-75-1, TMX2-28) (A) and EpCAMlow/neg cells (MDA-MB-231) (B) were either stained with 1 μM MitoTracker Green FM (A) or MitoTracker Orange CM (B) and incubated for cell adhesion experiments on NEXTERION slides AL, coated with different antibodies and ECM molecules, alone and in combination (0.1 mg/ml each). The labeling above the single spots indicates respective capture molecules (Iso = isotype control, ms = mouse, Lam = laminin, Col = collagen, HA = hyaluronic acid, rt = rat, T = Trop2, EpC = EpCAM, 49f = CD49f, rb = rabbit). (C) Overlay image of (A) and (B); scale bars (white) = 500 μm, 20x magnification. (D) Array layout (5x5 mm) with 36 spots (spot diameter = 500 μm; pitch = 800 μm) printed on NEXTERION slides AL. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26695635), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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); 397042 (Porcine); 478552 (Canine); 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; MER6; MER6integrin-associated signal transducer; OA3; Protein MER6; Rh-related antigen

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Citations for Human CD47 Antibody

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.

14 Citations: Showing 1 - 10
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  1. Remote Co-loading of amphipathic acid drugs in neutrophil nanovesicles infilled with cholesterol mitigates lung bacterial infection and inflammation
    Authors: Jin Gao, Yujie Su, Zhenjia Wang
    Biomaterials
  2. EGFR‐Induced and c‐Src‐Mediated CD47 Phosphorylation Inhibits TRIM21‐Dependent Polyubiquitylation and Degradation of CD47 to Promote Tumor Immune Evasion
    Authors: Linyong Du, Zhipeng Su, Silu Wang, Ying Meng, Fei Xiao, Daqian Xu et al.
    Advanced Science
  3. Senescent cells suppress macrophage-mediated corpse removal via upregulation of the CD47-QPCT/L axis
    Authors: Daniela Schloesser, Laura Lindenthal, Julia Sauer, Kyoung-Jin Chung, Triantafyllos Chavakis, Eva Griesser et al.
    Journal of Cell Biology
  4. Human lung adenocarcinoma CD47 is upregulated by interferon-gamma and promotes tumor metastasis
    Authors: Shuang Qu, Zichen Jiao, Geng Lu, Jiahan Xu, Bing Yao, Ting Wang et al.
    Molecular Therapy - Oncolytics
  5. Cancer cells under immune attack acquire CD47-mediated adaptive immune resistance independent of the myeloid CD47-SIRP alpha axis
    Authors: Mark A.J.M. Hendriks, Isabel Britsch, Xiurong Ke, Anne P. van Wijngarden, Douwe F. Samplonius, Emily M. Ploeg et al.
    OncoImmunology
  6. HIV-1 Vpu Promotes Phagocytosis of Infected CD4 + T Cells by Macrophages through Downregulation of CD47
    Authors: Lijun Cong, Scott M. Sugden, Pascal Leclair, Chinten James Lim, Tram N. Q. Pham, Éric A. Cohen
    mBio
  7. Bispecific antibody approach for EGFR-directed blockade of the CD47-SIRP alpha “don’t eat me” immune checkpoint promotes neutrophil-mediated trogoptosis and enhances antigen cross-presentation
    Authors: Mark A. J. M. Hendriks, Emily M. Ploeg, Iris Koopmans, Isabel Britsch, Xiurong Ke, Douwe F. Samplonius et al.
    OncoImmunology
  8. Formation of Human Neuroblastoma in Mouse-Human Neural Crest Chimeras
    Authors: MA Cohen, S Zhang, S Sengupta, H Ma, GW Bell, B Horton, B Sharma, RE George, S Spranger, R Jaenisch
    Cell Stem Cell, 2020-03-05;0(0):.
    Species: Mouse
    Sample Types: Tissue
    Applications: IHC-P
  9. Clinical Relevance of Immune Checkpoints on Circulating Tumor Cells in Breast Cancer
    Authors: MA Papadaki, AV Koutsopoul, PG Tsoulfas, E Lagoudaki, D Aggouraki, A Monastirio, C Koutoulaki, CA Apostolopo, AC Merodoulak, C Papadaki, D Mavroudis, S Agelaki
    Cancers (Basel), 2020-02-06;12(2):.
    Species: Human
    Sample Types: Whole Cells, Whole Tissue
    Applications: ICC, IHC
  10. Thrombospondin-1/CD47 Interaction Regulates Th17 and Treg Differentiation in Psoriasis
    Authors: Pedro Rodríguez-Jiménez, Pablo Chicharro, Mar Llamas-Velasco, Danay Cibrian, Laura Trigo-Torres, Alicia Vara et al.
    Frontiers in Immunology
  11. Combined prognostic value of the cancer stem cell markers CD47 and CD133 in esophageal squamous cell carcinoma
    Authors: JH Wang, ST Huang, L Zhang, ZG Liu, RX Liang, SW Jiang, YN Jiang, XJ Yu, YC Jiang, XZ Li, PF Zhang, ZS Wen, M Zheng
    Cancer Med, 2019-02-11;0(0):.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  12. 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, 2016-06-20;0(0):.
    Species: Human
    Sample Types: Whole Cells, Whole Tissue
    Applications: IHC-P, Neutralization
  13. EpCAM-Independent Enrichment of Circulating Tumor Cells in Metastatic Breast Cancer
    Authors: Helen Schneck, Berthold Gierke, Frauke Uppenkamp, Bianca Behrens, Dieter Niederacher, Nikolas H. Stoecklein et al.
    PLOS ONE
  14. 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, 2013-04-21;31(6):539-44.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P

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Human CD47 Antibody
By Anonymous on 05/20/2019
Application: WB Sample Tested: A549 human lung carcinoma cell line Species: Human

Human CD47 Antibody
By Anonymous on 10/17/2018
Application: WB Sample Tested: Breast cancer cells Species: Human

Human CD47 Antibody
By Anonymous on 07/15/2018
Application: WB Sample Tested: HepG2 human hepatocellular carcinoma cell line Species: Human