Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR10583] to TRIM56 - BSA and Azide free
  • Suitable for: ICC/IF, IHC-P, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-TRIM56 antibody [EPR10583] - BSA and Azide free
    See all TRIM56 primary antibodies

  • Description

    Rabbit monoclonal [EPR10583] to TRIM56 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IHC-P, Flow Cyt, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • IHC-P: Human brain, pancreas and colon cancer tissues. WB: HAP1, A549, A375, MCF7 and HeLa cell lysates. ICC/IF: MCF7 and HeLa cells Flow Cyt: MCF7 cells

  • General notes

    Ab249161 is the carrier-free version of ab154862. 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

    ab249161 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Images

  • Western blot - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)
    Western blot - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)
    All lanes : Anti-TRIM56 antibody [EPR10583] (ab154862) at 1/1000 dilution

    Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
    Lane 2 : TRIM56 knockout A549 (Human lung carcinoma cell line) whole cell lysate
    Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
    Lane 4 : A-375 (Human malignant melanoma cell line) whole 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: 81 kDa
    Observed band size: 88 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab154862, the same antibody clone in a different buffer formulation.

    Lanes 1-4: Merged signal (red and green). Green - ab154862 observed at 88 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab154862 Anti-TRIM56 antibody [EPR10583] was shown to specifically react with TRIM56 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267063 (knockout cell lysate ab258250) was used. Wild-type and TRIM56 knockout samples were subjected to SDS-PAGE. ab154862 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-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)
    Immunocytochemistry/ Immunofluorescence - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling TRIM56 with purified ab154862 at 1/70 dilution (10 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 µg/ml) dilution. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab154862).

  • Flow Cytometry - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)
    Flow Cytometry - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)

    Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling TRIM56 with purified ab154862 at 1/70 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab154862).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon cancer tissue sections labeling TRIM56 with purified ab154862 at 1/1000 dilution (0.691 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab154862).

  • Western blot - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)
    Western blot - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)
    All lanes : Anti-TRIM56 antibody [EPR10583] (ab154862) at 1/1000 dilution

    Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
    Lane 2 : TRIM56 knockout A549 (Human lung carcinoma cell line) whole cell lysate
    Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
    Lane 4 : A-375 (Human malignant melanoma cell line) whole 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: 81 kDa
    Observed band size: 88 kDa why is the actual band size different from the predicted?



    This data was developed using ab154862, the same antibody clone in a different buffer formulation.

    Lanes 1-4: Merged signal (red and green). Green - ab154862 observed at 88 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab154862 Anti-TRIM56 antibody [EPR10583] was shown to specifically react with TRIM56 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267062 (knockout cell lysate ab258249) was used. Wild-type and TRIM56 knockout samples were subjected to SDS-PAGE. ab154862 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.

  • Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)
    Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (ab249161)

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