Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16792] to PML Protein - BSA and Azide free
  • Suitable for: WB, ICC/IF, IP, IHC-P
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-PML Protein antibody [EPR16792] - BSA and Azide free
    See all PML Protein primary antibodies

  • Description

    Rabbit monoclonal [EPR16792] to PML Protein - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: 293T, K562, HeLa and A549 whole cell lysates; Human fetal brain, fetal heart and fetal kidney lysates; HAP1. IHC-P: Human mammary gland and breast carcinoma tissues. ICC/IF: K562 cells. ICC/IF KO: Hap1 cells (Hap1-PML KO used as a negative cell line). IP: K562 whole cell extract.

  • General notes

    Ab240213 is the carrier-free version of ab179466. 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

    ab240213 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.

    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

Images

  • Western blot - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)
    Western blot - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)
    All lanes : Anti-PML Protein antibody [EPR16792] (ab179466) at 1/1000 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : PML knockout A549 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 98 kDa
    Observed band size: 110 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 (ab179466).

    Lanes 1 - 2: Merged signal (red and green). Green - ab179466 observed at 110 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

    ab179466 was shown to react with PML in A549 wild-type cells in western blot with loss of signal observed in PML knockout cell line ab266980 (PML knockout cell lysate ab257082). Wild-type and PML knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk before incubation with ab179466 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)
    Immunocytochemistry/ Immunofluorescence - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling PML Protein with ab179466 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm and nuclear staining on K562 cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

    The negative controls are as follows:-
    -ve control 1 -  ab179466 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179466).

  • Immunoprecipitation - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)
    Immunoprecipitation - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)

    PML Protein was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell extract with ab179466 at 1/70 diltuion. Western blot was performed from the immunoprecipitate using ab179466 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: K562 whole cell extract. Lane 2: PBS instead of K562 whole cell extract.

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

    ab179466 recognizes 12 isoforms shown as multiple bands ranging from 50 kDa to 110 kDa.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179466).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)

    Immunohistochemical analysis of paraffin-embedded Human mammary gland tissue labeling PML Protein with ab179466 at 1/2000 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus staining on epithelial cells of Human mammary gland tissue is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179466).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)
    Western blot - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)
    All lanes : Anti-PML Protein antibody [EPR16792] (ab179466) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : PML knockout HeLa cell lysate
    Lane 3 : 293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 98 kDa
    Observed band size: 110 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 (ab179466).

    Lanes 1-3: Merged signal (red and green). Green - ab179466 observed at 50-110 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab179466 Anti-PML Protein antibody [EPR16792] was shown to specifically react with PML Protein in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261811 (knockout cell lysate ab257081) was used. Wild-type and PML Protein knockout samples were subjected to SDS-PAGE. ab179466 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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 10000 dilution for 1 hour at room temperature before imaging.

     

     

  • Western blot - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)
    Western blot - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)
    All lanes : Anti-PML Protein antibody [EPR16792] (ab179466) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : PML knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : HEK293 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 98 kDa
    Observed band size: 50-110 kDa why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab179466 observed at 50-110 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab179466 was shown to recognize in wild-type HAP1 cells as signal was lost at the expected MW in PML knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PML knockout samples were subjected to SDS-PAGE. Ab179466 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179466).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)

    Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling PML Protein with ab179466 at 1/2000 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with Hematoxylin. The breast cancer cells lost expression.

    Reference: Gurrieri C et al. Loss of the Tumor Suppressor PML in Human Cancers of Multiple Histologic Origins. J Natl Cancer Inst 96:269 –279 (2004).

    Negative control: Used PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179466).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)
    Anti-PML Protein antibody [EPR16792] - BSA and Azide free (ab240213)

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