Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3947] to PMS2 - BSA and Azide free
  • Suitable for: Flow Cyt, ICC/IF, IHC-P, WB, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-PMS2 antibody [EPR3947] - BSA and Azide free
    See all PMS2 primary antibodies

  • Description

    Rabbit monoclonal [EPR3947] to PMS2 - 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: Hap1 and HeLa cell lysates. Flow Cyt: HeLa cells. ICC/IF: HeLa cells. IHC-P: Human colonic adenocarcinoma tissue. IP: HeLa whole cell lysate.

  • General notes

    Ab214442 is the carrier-free version of ab110638. 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

    ab214442 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)

    This IHC data was generated using the same anti-PMS2 antibody clone, EPR3947, in a different buffer formulation (cat# ab110638).

    ab110638 at 1/100 dilution staining PMS2 in Human colonic adenocarcinoma by Immunohistochemistry, Paraffin-embedded tissue.

    Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    Western blot - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    All lanes : Anti-PMS2 antibody [EPR3947] (ab110638) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : PMS2 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 96 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 (ab110638).

      Lanes 1- 2: Merged signal (red and green). Green - ab110638 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab110638 was shown to react with PMS2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261776 (knockout cell lysate ab257142) was used. Wild-type HeLa and PMS2 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. ab110638 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    Immunocytochemistry/ Immunofluorescence - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    ab110638 at 1/100 dilution staining PMS2 in HeLa cells by Immunofluorescence.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab110638).

  • Immunoprecipitation - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    Immunoprecipitation - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)

    This data was developed using ab110638, the same antibody clone in a different buffer formulation.
    Purified ab110638 at 1/50 dilution (2µg) immunoprecipitating PMS2 in HeLa whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab110638 + HeLa whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab110638 in HeLa whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 110 kDa
    Lower bands are degradation bands and fresh lysate is recommended.

  • Flow Cytometry - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    Flow Cytometry - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)

    Clone EPR3947 (ab214442) has been successfully conjugated by Abcam. This image was generated using Anti-PMS2 antibody [EPR3947] (Alexa Fluor® 647). Please refer to ab202835 for protocol details.

    Overlay histogram showing HeLa cells stained with ab202835 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab202835, 1/1500 dilution) for 30 min at 22°C.

    Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 647 (ab199093 used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 40mW Red laser (640nm) and 670/14 bandpass filter.

    This antibody gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min)/ /permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

  • Western blot - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    Western blot - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    All lanes : Anti-PMS2 antibody [EPR3947] (ab110638) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : MS2 knockout HAP1 whole cell lysate

    Lysates/proteins at 30 µg per lane.

    Predicted band size: 96 kDa



    This WB data was generated using the same anti-PMS2 antibody clone, EPR3947, in a different buffer formulation (cat# ab110638).

    Lanes 1-2: Merged signal (red and green). Green - ab110638 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab110638 was shown to specifically react with PMS2 in wild-type HAP1 cells. No band was observed when PMS2 knockout samples were used. Wild-type and PMS2 knockout samples were subjected to SDS-PAGE.  Ab110638 and ab8245 (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-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    Flow Cytometry - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    Overlay histogram showing HeLa cells stained with ab110638 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab110638, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab110638).

  • Immunoprecipitation - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    Immunoprecipitation - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)

    This data was developed using ab110638, the same antibody clone in a different buffer formulation.
    Purified ab110638 at 1/50 dilution (2µg) immunoprecipitating PMS2 in HeLa whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab110638 + HeLa whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab110638 in HeLa whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 110 kDa
    Lysate were made freshly and used in IP test immediately to minimize protein degradation. Incubation time was 2h.

  • OI-RD Scanning - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    OI-RD Scanning - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab110638).

  • Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)
    Anti-PMS2 antibody [EPR3947] - BSA and Azide free (ab214442)

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