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Cardiovascular Cardiovascular Markers Cell Markers Endothelial Cells

Anti-CD146 antibody [EPR3208] (ab75769)

Price and availability

367 717 ₸

Availability

Order now and get it on Tuesday March 02, 2021

Anti-CD146 antibody [EPR3208] (ab75769)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3208] to CD146
  • Suitable for: ICC/IF, WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-CD146 antibody [EPR3208]
    See all CD146 primary antibodies
  • Description

    Rabbit monoclonal [EPR3208] to CD146
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human CD146 aa 600 to the C-terminus.
    Database link: P43121

  • Positive control

    • WB: HeLa, A375, HUVEC and B16-F0 cell lysate; Human fetal artery lysate and Rat placenta. ICC/IF: Murine bone marrow cell lysates. IHC-Fr: Mouse spleen tissue. IHC-P: Melanoma, breast carcinoma vessel, urinary bladder transitional carcinoma vessel, glioma vessel, normal tonsil and normal spleen tissue. Flow Cyt: A375 and HUVEC cells
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3208
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Cell Type Markers
    • CD
    • Endothelial Cells
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cell Adhesion
    • Cell Adhesion Molecules
    • Endothelial
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Cell adhesion
    • Other
    • Stem Cells
    • Endothelial Progenitors
    • Endothelial Markers
    • Cardiovascular
    • Angiogenesis
    • Endothelial Cell Markers

Images

  • Western blot - Anti-CD146 antibody [EPR3208] (ab75769)
    Western blot - Anti-CD146 antibody [EPR3208] (ab75769)

    Lane 1: Wild-type HAP1 whole cell lysate (40 µg)
    Lane 2: CD146 knockout HAP1 whole cell lysate (40 µg)
    Lane 3: A375 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab75769 observed at 120-72 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab75769 was shown to specifically react with CD146  in wild-type HAP1 cells as signal was lost in CD146 knockout cells. Wild-type and CD146 knockout samples were subjected to SDS-PAGE. ab75769 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769) Davaine, J.M. et al PLoS One. 2014 Sep 26;9(9):e107642. doi: 10.1371/journal.pone.0107642. eCollection 2014 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Immunohistochemistry experiments were used to compare symptomatic carotid plaques (SC) and asymptomatic carotid plaques (AsC)

    Asymptomatic lesions presented higher CD146+ pericyte infiltration, p

    ab75769 used at 1/200 dilution.

    (After Figure 2 of Davaine et al)

  • Immunocytochemistry/ Immunofluorescence - Anti-CD146 antibody [EPR3208] (ab75769)
    Immunocytochemistry/ Immunofluorescence - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunocytochemistry/Immunofluorescence analysis of A375 (human malignant melanoma) cells labelling CD146 with purified ab75769 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD146 with purified ab75769 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Flow Cytometry - Anti-CD146 antibody [EPR3208] (ab75769)
    Flow Cytometry - Anti-CD146 antibody [EPR3208] (ab75769)

    Flow Cytometry analysis of A375 (human malignant melanoma) cells labeling CD146 with unpurified ab75769 at 1/20 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • Western blot - Anti-CD146 antibody [EPR3208] (ab75769)
    Western blot - Anti-CD146 antibody [EPR3208] (ab75769)
    All lanes : Anti-CD146 antibody [EPR3208] (ab75769) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : MCAM knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 72 kDa
    Observed band size: 120 kDa
    why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab75769 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab75769 was shown to react with CD146 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab261790 (knockout cell lysate ab256985) lane below 120kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and MCAM knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab75769 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 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.

  • Western blot - Anti-CD146 antibody [EPR3208] (ab75769)
    Western blot - Anti-CD146 antibody [EPR3208] (ab75769)
    All lanes : Anti-CD146 antibody [EPR3208] (ab75769) at 1/10000 dilution (purified)

    Lane 1 : A375 cell lysate
    Lane 2 : Human fetal artery lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 72 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-CD146 antibody [EPR3208] (ab75769)
    Western blot - Anti-CD146 antibody [EPR3208] (ab75769)
    Anti-CD146 antibody [EPR3208] (ab75769) at 1/10000 dilution (purified) + HUVEC cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 72 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-CD146 antibody [EPR3208] (ab75769)
    Western blot - Anti-CD146 antibody [EPR3208] (ab75769)
    Anti-CD146 antibody [EPR3208] (ab75769) at 1/10000 dilution (purified) + B16-F0 cell lysate at 20 µg/ml

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 72 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-CD146 antibody [EPR3208] (ab75769)
    Western blot - Anti-CD146 antibody [EPR3208] (ab75769)
    Anti-CD146 antibody [EPR3208] (ab75769) at 1/10000 dilution (purified) + Rat placenta lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 72 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of melanoma tissue labelling CD146 with unpurified ab75769 at 1/250. A HRP/AP polymerized secondary antibody was used.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of breast carcinoma vessels tissue labelling CD146 with unpurified ab75769.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of urinary bladder transitional carcinoma vessels tissue labelling CD146 with unpurified ab75769.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of glioma vessels tissue labelling CD146 with unpurified ab75769.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal tonsil tissue labelling CD146 with unpurified ab75769.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal spleen tissue labelling CD146 with unpurified ab75769.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-CD146 antibody [EPR3208] (ab75769)
    Flow Cytometry - Anti-CD146 antibody [EPR3208] (ab75769)

    Flow Cytometry analysis of HUVEC cells labelling CD146 with purified ab75769 at 1/50 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Anti-CD146 antibody [EPR3208] (ab75769)
    Anti-CD146 antibody [EPR3208] (ab75769)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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