Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2759(2)] to CD166 - BSA and Azide free
  • Suitable for: WB, IP, ICC/IF, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free
    See all CD166 primary antibodies

  • Description

    Rabbit monoclonal [EPR2759(2)] to CD166 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

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

  • Immunogen

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

  • Positive control

    • WB: SH-SY5Y, HuT-78, HT-1080 and Daudi cell lysates. Mouse and rat brain tissue lysates. IHC-P: Human liver and prostatic adenocarcinoma tissues. ICC/IF: Jurkat cells. Flow Cyt: HuT-78 cells. IP: SH-SY5Y cells.

  • General notes

    Ab206127 is the carrier-free version of ab109215. 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

    ab206127 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.

    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.

    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

  • Flow Cytometry - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)
    Flow Cytometry - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)

    Flow Cytometry analysis of HuT-78 cells labelling CD166 with purified ab109215 at 1/90 (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 (ab109215).

  • Immunocytochemistry/ Immunofluorescence - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)
    Immunocytochemistry/ Immunofluorescence - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)

    Immunocytochemistry/Immunofluorescence analysis of THP-1 (human monocytic leukemia cell line) cells labelling CD166 (green) with purified ab109215 at 1/250. Cells were fixed with 100% methanol. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as a nuclear counterstain.  

    Secondary Only Control: PBS was used instead of the primary antibody as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109215).

  • Immunoprecipitation - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)
    Immunoprecipitation - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)

    ab109215 (purified) at 1/30 immunoprecipitating CD166 in SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109215).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling CD166 with purified ab109215 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109215).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling CD166 with unpurified ab109215 at 1/100.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109215).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic adenocarcinoma tissue labelling CD166 with unpurified ab109215 at 1/100.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109215).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)
    Western blot - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109215).

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: CD166 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: SH-SY5Y whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab109215 observed at 100 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab109215 was shown to specifically react with CD166 in wild-type HAP1 cells as signal was lost in CD166 knockout cells. Wild-type and CD166 knockout samples were subjected to SDS-PAGE. Ab109215 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)
    Western blot - Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)
    Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127) + SH-SY5Y (human neuroblastoma) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

    Predicted band size: 65 kDa
    Observed band size: 100-105 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM/TBST

  • Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)
    Anti-CD166 antibody [EPR2759(2)] - BSA and Azide free (ab206127)

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