Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PICALM antibody [EPR12177] - BSA and Azide free (ab249736)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR12177] to PICALM - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-PICALM antibody [EPR12177] - BSA and Azide free
    See all PICALM primary antibodies

  • Description

    Rabbit monoclonal [EPR12177] to PICALM - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt or IP

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse

  • Immunogen

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

  • Positive control

    • WB: HeLa, U-2 OS, PC-3, and Caco-2 cell lysates.

  • General notes

    ab249736 is the carrier-free version of ab172962. 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

    ab249736 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.

Properties

Images

  • Western blot - Anti-PICALM antibody [EPR12177] - BSA and Azide free (ab249736)
    Western blot - Anti-PICALM antibody [EPR12177] - BSA and Azide free (ab249736)
    All lanes : Anti-PICALM antibody [EPR12177] (ab172962) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : PICALM knockout HeLa cell lysate
    Lane 3 : Caco-2 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 71 kDa
    Observed band size: 70-75 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 (ab172962).

    Lanes 1-3: Merged signal (red and green). Green - ab172962 observed at 70-75 kDa. Red - loading control ab8245 observed at 36 kDa.

    ab172962 Anti-PICALM antibody [EPR12177] was shown to specifically react with PICALM in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265653 (knockout cell lysate ab258113) was used. Wild-type and PICALM knockout samples were subjected to SDS-PAGE. ab172962 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 20000 dilution for 1 hour at room temperature before imaging.

     

     

  • Western blot - Anti-PICALM antibody [EPR12177] - BSA and Azide free (ab249736)
    Western blot - Anti-PICALM antibody [EPR12177] - BSA and Azide free (ab249736)
    All lanes : Anti-PICALM antibody [EPR12177] (ab172962) at 1/5000 dilution

    Lane 1 : Wild-type U-2 OS whole cell lysate
    Lane 2 : PICALM knockout U-2 OS whole cell lysate
    Lane 3 : PC-3 whole cell lysate
    Lane 4 : Caco-2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 71 kDa
    Observed band size: 70-75 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 (ab172962).

    Lanes 1 - 4: Merged signal (red and green). Green - ab172962 observed at 70-75 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

    ab172962 was shown to react with PICALM in U-2 OS wild-type cells in Western blot. Loss of signal was observed when PICALM knockout sample was used. U-2 OS wild-type and PICALM knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab172962 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 5000 dilution and a 1 in 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Anti-PICALM antibody [EPR12177] - BSA and Azide free (ab249736)
    Anti-PICALM antibody [EPR12177] - BSA and Azide free (ab249736)

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