Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)

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 - BSA and Azide free
  • Suitable for: WB, Flow Cyt, ICC/IF, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-CD146 antibody [EPR3208] - BSA and Azide free
    See all CD146 primary antibodies

  • Description

    Rabbit monoclonal [EPR3208] to CD146 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

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

  • Immunogen

    This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • ICC/IF: A375 cells. IHC-Fr: Mouse spleen tissue. IHC-P: Melanoma, breast carcinoma vessel, urinary bladder transitional carcinoma vessel, glioma vessel, normal tonsil and normal spleen tissue. WB: HeLa and HAP1 cell lysates. Flow Cyt: A375 and HUVEC cells.

  • General notes

    Ab210072 is the carrier-free version of ab75769. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab210072 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
    Western blot - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).

  • Flow Cytometry - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
    Flow Cytometry - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).

  • Immunocytochemistry/ Immunofluorescence - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
    Immunocytochemistry/ Immunofluorescence - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).

  • Western blot - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
    Western blot - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
    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?



    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)

    This IHC data was generated using the same anti-CD146 antibody clone, EPR3208, in a different buffer formulation (cat# 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072) Image from Davaine J.M. et al. PLoS One. 2014 Sep 26;9(9):e107642. doi: 10.1371/journal.pone.0107642. eCollection 2014.

    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)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).

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

  • Flow Cytometry - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
    Flow Cytometry - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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] - BSA and Azide free (ab210072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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] - BSA and Azide free (ab210072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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] - BSA and Azide free (ab210072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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] - BSA and Azide free (ab210072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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] - BSA and Azide free (ab210072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).

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

  • Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
    Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)

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