Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-KAP1 (phospho S824) antibody [EPR5248] - BSA and Azide free (ab215549)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5248] to KAP1 (phospho S824) - BSA and Azide free
  • Suitable for: WB, IP, Dot blot
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-KAP1 (phospho S824) antibody [EPR5248] - BSA and Azide free
    See all KAP1 primary antibodies

  • Description

    Rabbit monoclonal [EPR5248] to KAP1 (phospho S824) - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    ab133440 only detects KAP1 phosphorylated at Serine 824.

  • Tested Applications & Species

    Application Species
    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: WT HAP1 cell lysate (+/- Bleomycin); HeLa cell lysate (+/- Bleomycin). Dot blot: KAP1 (phospho S824) phospho peptide. IP: HeLa treated with 3uM etoposide for 1h whole cell lysate.

  • General notes

    ab215549 is the carrier-free version of ab133440 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

    Ab215549 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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.

Properties

Images

  • Western blot - Anti-KAP1 (phospho S824) antibody [EPR5248] - BSA and Azide free (ab215549)
    Western blot - Anti-KAP1 (phospho S824) antibody [EPR5248] - BSA and Azide free (ab215549)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2:     Wild type HAP1 + DMSO whole cell lysate (20 µg)
    Lane 3:     Wild type HAP1 + Bleomycin whole cell lysate (20 µg)
    Lane 4:     KAP1 knockout HAP1 whole cell lysate (20 µg)
    Lane 5:     KAP1 knockout HAP1 + DMSO whole cell lysate (20 µg)
    Lane 6:     KAP1 knockout HAP1 + Bleomycin whole cell lysate (20 µg)
    Lane 7:     HeLa + DMSO whole cell lysate (20 µg)
    Lane 8: HeLa + Bleomycin whole cell lysate (20 µg)

    Lanes 1 - 8: Merged signal (red and green). Green - ab133440 observed at 105 kDa. Red - loading control, ab130007, observed at 125 kDa.

    ab133440 was shown to specifically react with KAP1 in wild type cells as signal was lost in KAP1 knockout cells. Wild-type and KAP1 knockout samples were subjected to SDS-PAGE. ab133440 and ab130007 (Mouse anti-vinculin loading control) were incubated overnight at 4°C both at a 1/20000 dilution. 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. Treated with 30 µg/mL Bleomycin in DMSO for 30 minutes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133440).

  • Immunoprecipitation - Anti-KAP1 (phospho S824) antibody [EPR5248] - BSA and Azide free (ab215549)
    Immunoprecipitation - Anti-KAP1 (phospho S824) antibody [EPR5248] - BSA and Azide free (ab215549)

    This data was developed using ab133440, the same antibody clone in a different buffer formulation.
    Purified ab133440 at 1/50 dilution (2µg) immunoprecipitating KAP1 in HeLa treated with 3uM etoposide for 1h whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) treated with 3uM etoposide for 1h whole cell lysate 10µg
    Lane 2 (+): ab133440 + HeLa treated with 3uM etoposide for 1h whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32132 in HeLa treated with 3uM etoposide for 1h whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 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

  • Dot Blot - Anti-KAP1 (phospho S824) antibody [EPR5248] - BSA and Azide free (ab215549)
    Dot Blot - Anti-KAP1 (phospho S824) antibody [EPR5248] - BSA and Azide free (ab215549)

    Dot blot analysis of KAP1 (phospho S824) phospho peptide (Lane 1) and KAP1 non-phospho peptide (Lane 2) labelling KAP1 (phospho S824) with ab133440 at a dilution of 1/1000. A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/20,000. Blocking buffer: 5% NFDM/TBST. Dilution buffer: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133440)
  • Anti-KAP1 (phospho S824) antibody [EPR5248] - BSA and Azide free (ab215549)
    Anti-KAP1 (phospho S824) antibody [EPR5248] - BSA and Azide free (ab215549)

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