Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (ab247421)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2520Y] to PBK/SPK - BSA and Azide free
  • Suitable for: IHC-P, WB, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free
    See all PBK/SPK primary antibodies

  • Description

    Rabbit monoclonal [EP2520Y] to PBK/SPK - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WB, IPmore details
    Unsuitable for: ICC

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: Human tonsil tissue lysate; HEK-293T, HeLa and HAP1 cell lysates. IHC-P: Human tonsil tissue

  • General notes

    ab247421 is the carrier-free version of ab75987 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

    Ab247421 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 was previously labelled as PBK

    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-PBK/SPK antibody [EP2520Y] - BSA and Azide free (ab247421)
    Western blot - Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (ab247421)
    All lanes : Anti-PBK/SPK antibody [EP2520Y] (ab75987) at 1/100000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : PBK knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 36 kDa
    Observed band size: 40 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 (ab75987).

    Lanes 1- 2: Merged signal (red and green). Green - ab75987 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab75987 was shown to react with PBK/SPK in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266827 (knockout cell lysate ab257575) was used. Wild-type HEK-293T and PBK knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab75987 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 100000 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-PBK/SPK antibody [EP2520Y] - BSA and Azide free (ab247421)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (ab247421)

    Immunohistochemical analysis of paraffin-embedded human tonsil using ab75987 at 1/100 dilution. Antigen retrieval was heat mediated with citrate buffer pH 6 before commencing with IHC staining protocol. 

    This data was developed using the same antibody clone in a different buffer formulation (ab75987).

  • Western blot - Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (ab247421)
    Western blot - Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (ab247421)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: PBK knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: Human tonsil tissue lysate (20 µg) 
    Lanes 1 - 4: Merged signal (red and green). Green - ab75987 observed at 40 kDa. Red - loading control, ab7291, observed at 52 kDa.

    ab75987 was shown to recognize PBK when PBK knockout samples were used, along with additional cross-reactive bands. Wild-type and PBK knockout samples were subjected to SDS-PAGE. ab75987 diluted to 1/100,000 and ab7291 (loading control to alpha Tubulin) diluted to 1/10,000 and incubated overnight at 4°C. 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 1/10,000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation (ab75987).

  • Western blot - Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (ab247421)
    Western blot - Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (ab247421)
    Anti-PBK/SPK antibody [EP2520Y] (ab75987) at 1/500000 dilution + HeLa cell lysate at 10 µg

    Secondary
    HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 36 kDa
    Observed band size: 40 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 (ab75987).

  • Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (ab247421)
    Anti-PBK/SPK antibody [EP2520Y] - BSA and Azide free (ab247421)

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