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    cd147-antibody-10e10-ab230921.pdf

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Immunology Cell Type Markers CD Endothelial Cells
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Validated using a knockout cell line

Anti-CD147 antibody [10E10] (ab230921)

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Western blot - Anti-CD147 antibody [10E10] (ab230921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [10E10] (ab230921)
  • Western blot - Anti-CD147 antibody [10E10] (ab230921)
  • Flow Cytometry - Anti-CD147 antibody [10E10] (ab230921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [10E10] (ab230921)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD147 antibody [10E10] (ab230921)
  • Western blot - Anti-CD147 antibody [10E10] (ab230921)
  • Flow Cytometry - Anti-CD147 antibody [10E10] (ab230921)
  • Flow Cytometry - Anti-CD147 antibody [10E10] (ab230921)

Key features and details

  • Mouse monoclonal [10E10] to CD147
  • Suitable for: WB, Flow Cyt, ICC/IF, IHC-P
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG2b

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Knockout
Product image
Human BSG (CD147) knockout A549 cell line (ab273748)
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PE Anti-CD147 antibody [MEM-M6/1] (ab77133)

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Overview

  • Product name

    Anti-CD147 antibody [10E10]
    See all CD147 primary antibodies
  • Description

    Mouse monoclonal [10E10] to CD147
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Rat, Dog, Monkey
  • Immunogen

    Recombinant full length protein corresponding to Human CD147. (NP_001719). Produced in HEK293T cell.
    Database link: P35613

  • Positive control

    • WB: HEK-293T, HepG2, HeLa, HT-29, A549, COS-7, Jurkat, MDCK, Raji, Jurkat, PC-12 and MCF7 whole cell lysate. IHC-P: Human brain and placenta tissue. Flo Cyt: A549, HeLa and Jurkat cells. ICC: HeLa cells.
  • General notes

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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: 7.30
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 1% BSA, 50% Glycerol
  • Concentration information loading...
  • Purity

    Protein A/G purified
  • Clonality

    Monoclonal
  • Clone number

    10E10
  • Isotype

    IgG2b
  • Research areas

    • Immunology
    • Cell Type Markers
    • CD
    • Endothelial Cells
    • Neuroscience
    • Sensory System
    • Visual system
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Extracellular matrix
    • Other
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Cardiovascular
    • Angiogenesis
    • Endothelial Cell Markers

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
  • KO cell lines

    • Human BSG (CD147) knockout A549 cell line (ab273748)
  • KO cell lysates

    • Human BSG (CD147) knockout A549 cell lysate (ab275500)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HepG2 whole cell lysate (ab166833)
    • MCF7 whole cell lysate (ab29537)
    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab30128)
    • HT29 whole cell lysate (ab3952)
    • Jurkat whole cell lysate (ab7899)
    • A549 whole cell lysate (ab7910)

Applications

Our Abpromise guarantee covers the use of ab230921 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 1/500 - 1/2000.
Flow Cyt 1/100.
ICC/IF 1/100.
IHC-P Use a concentration of 10 µg/ml.

Target

  • Function

    Plays pivotal roles in spermatogenesis, embryo implantation, neural network formation and tumor progression. Stimulates adjacent fibroblasts to produce matrix metalloproteinases (MMPS). May target monocarboxylate transporters SLC16A1, SLC16A3 and SLC16A8 to plasma membranes of retinal pigment epithelium and neural retina. Seems to be a receptor for oligomannosidic glycans. In vitro, promotes outgrowth of astrocytic processes.
  • Tissue specificity

    Present only in vascular endothelium in non-neoplastic regions of the brain, whereas it is present in tumor cells but not in proliferating blood vessels in malignant gliomas.
  • Sequence similarities

    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Post-translational
    modifications

    N-glycosylated.
  • Cellular localization

    Cell membrane. Melanosome. Colocalizes with SLC16A1 and SLC16A8 (By similarity). Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Target information above from: UniProt accession P35613 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 682 Human
    • Entrez Gene: 25246 Rat
    • Omim: 109480 Human
    • SwissProt: P35613 Human
    • SwissProt: P26453 Rat
    • Unigene: 501293 Human
    • Unigene: 2269 Rat
    • Alternative names

      • 5A11 antigen antibody
      • 5F7 antibody
      • BASI_HUMAN antibody
      • Basigin (Ok blood group) antibody
      • Basigin antibody
      • Blood brain barrier HT7 antigen antibody
      • Bsg antibody
      • CD 147 antibody
      • CD147 antibody
      • CD147 antigen antibody
      • Collagenase stimulatory factor antibody
      • EMMPRIN antibody
      • Extracellular matrix metalloproteinase inducer antibody
      • Leukocyte activation antigen M6 antibody
      • M 6 antibody
      • M6 antibody
      • M6 leukocyte activation antigen antibody
      • Neurothelin antibody
      • OK antibody
      • OK blood group antibody
      • OK blood group antigen antibody
      • TCSF antibody
      • Tumor cell derived collagenase stimulatory factor antibody
      • Tumor cell-derived collagenase stimulatory factor antibody
      see all

    Images

    • Western blot - Anti-CD147 antibody [10E10] (ab230921)
      Western blot - Anti-CD147 antibody [10E10] (ab230921)
      All lanes : Anti-CD147 antibody [10E10] (ab230921) at 1/500 dilution

      Lane 1 : pCMV6-ENTRY control-transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
      Lane 2 : pCMV6-ENTRY CD147-transfected HEK-293T whole cell lysate

      Lysates/proteins at 5 µg per lane.

      Developed using the ECL technique.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [10E10] (ab230921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [10E10] (ab230921)

      Formalin-fixed, paraffin-embedded human brain tissue stained for CD147 using ab230921 at 10 μg/ml in immunohistochemical analysis.

    • Western blot - Anti-CD147 antibody [10E10] (ab230921)
      Western blot - Anti-CD147 antibody [10E10] (ab230921)
      All lanes : Anti-CD147 antibody [10E10] (ab230921) at 1/500 dilution

      Lane 1 : Wild-type A549 cell lysate
      Lane 2 : BSG knockout A549 cell lysate
      Lane 3 : Raji cell lysate
      Lane 4 : Jurkat cell lysate

      Lysates/proteins at 30 µg per lane.

      Performed under reducing conditions.

      Observed band size: 42-70 kDa
      why is the actual band size different from the predicted?



      Lanes 1 - 4: Merged signal (red and green). Green - ab230921 observed at 42-70 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

      ab230921 was shown to react with CD147 in wild-type A549 cells in western blot with loss of signal observed in BSG knockout cell line ab273748 (knockout cell lysate ab275500). Wild-type and BSG knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab230921 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at a 1 in 500 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Flow Cytometry - Anti-CD147 antibody [10E10] (ab230921)
      Flow Cytometry - Anti-CD147 antibody [10E10] (ab230921)

      Flow cytometry overlay histogram showing wild-type A549 (green line) and BSG knockout A549 cells (ab273748) stained with ab230921 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab230921) (1x106 in 100μl at 5 μg/ml) for 30 min at 4°C.

      The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150117) was used at 1/2000 for 30 min at 4°C.

      Isotype control antibody was mouse IgG1κ (ab170190) used at the same concentration and conditions as the primary antibody (wild-type A549 - black line; BSG knockout A549 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

      Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [10E10] (ab230921)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [10E10] (ab230921)

      Formalin-fixed, paraffin-embedded human placenta tissue stained for CD147 using ab230921 at 10 μg/ml in immunohistochemical analysis.

    • Immunocytochemistry/ Immunofluorescence - Anti-CD147 antibody [10E10] (ab230921)
      Immunocytochemistry/ Immunofluorescence - Anti-CD147 antibody [10E10] (ab230921)

      HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for CD147 (green) using ab230921 at 1/100 dilution in ICC/IF.

    • Western blot - Anti-CD147 antibody [10E10] (ab230921)
      Western blot - Anti-CD147 antibody [10E10] (ab230921)
      All lanes : Anti-CD147 antibody [10E10] (ab230921) at 1/500 dilution

      Lane 1 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
      Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 3 : HT-29 (human colorectal adenocarcinoma cell line) whole cell lysate
      Lane 4 : A549 (human lung carcinoma cell line) whole cell lysate
      Lane 5 : COS-7 (African green monkey kidney fibroblast-like cell line) whole cell lysate
      Lane 6 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
      Lane 7 : MDCK (canine kidney cell line) whole cell lysate
      Lane 8 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
      Lane 9 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate

      Lysates/proteins at 35 µg per lane.

      Developed using the ECL technique.
    • Flow Cytometry - Anti-CD147 antibody [10E10] (ab230921)
      Flow Cytometry - Anti-CD147 antibody [10E10] (ab230921)

      Flow cytometric analysis of HeLa (human T cell leukemia cell line from peripheral blood) cell line labeling CD147 with ab230921 at 1/100 dilution (red) compared to a nonspecific negative control antibody (blue).

    • Flow Cytometry - Anti-CD147 antibody [10E10] (ab230921)
      Flow Cytometry - Anti-CD147 antibody [10E10] (ab230921)

      Flow cytometric analysis of Jurkat (human T cell leukemia cell line from peripheral blood) cell line labeling CD147 with ab230921 at 1/100 dilution (red) compared to a nonspecific negative control antibody (blue).

    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
  • References (0)

    Publishing research using ab230921? Please let us know so that we can cite the reference in this datasheet.

    ab230921 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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