Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PICALM antibody [EPR12177] (ab172962)

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
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-PICALM antibody [EPR12177]
    See all PICALM primary antibodies

  • Description

    Rabbit monoclonal [EPR12177] to PICALM

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human PICALM aa 550-650. The exact sequence is proprietary.
    Database link: Q13492

  • Positive control

    • WB: U-2 OS, PC-3, Caco-2, HepG2, HeLa, and A431 cell lysates and human fetal brain tissue lysates. IHC-P: Human placenta and breast tissues. ICC/IF: HeLa cells.

  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this 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-PICALM antibody [EPR12177] (ab172962)
    Western blot - Anti-PICALM antibody [EPR12177] (ab172962)
    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?



    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.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-PICALM antibody [EPR12177] (ab172962)
    Immunocytochemistry/ Immunofluorescence - Anti-PICALM antibody [EPR12177] (ab172962)

    Immunofluorescent analysis of HeLa cells labeling PICALM with ab172962 at 1/100 dilution.

  • Western blot - Anti-PICALM antibody [EPR12177] (ab172962)
    Western blot - Anti-PICALM antibody [EPR12177] (ab172962)
    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?



    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.

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

    Lane 1 : HepG2 lysate
    Lane 2 : HeLa lysate
    Lane 3 : A431 lysate
    Lane 4 : Human fetal brain lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size: 71 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PICALM antibody [EPR12177] (ab172962)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PICALM antibody [EPR12177] (ab172962)

    Immunohistochemical analysis of paraffin-embedded Human Placenta tissue labeling PICALM with ab172962 at 1/50 dilution.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PICALM antibody [EPR12177] (ab172962)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PICALM antibody [EPR12177] (ab172962)

    Immunohistochemical analysis of paraffin-embedded Human Breast tissue labeling PICALM with ab172962 at 1/50 dilution.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Anti-PICALM antibody [EPR12177] (ab172962)
    Anti-PICALM antibody [EPR12177] (ab172962)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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