Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR10582] to TRIM56 - BSA and Azide free
  • Suitable for: ICC, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-TRIM56 antibody [EPR10582] - BSA and Azide free
    See all TRIM56 primary antibodies

  • Description

    Rabbit monoclonal [EPR10582] to TRIM56 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Human

  • Positive control

    • WB: A549, MCF-7, HeLa and A375 cell lysates. ICC: MCF-7 cells. Flow Cyt: HeLa cells.

  • General notes

    Ab249127 is the carrier-free version of ab154821. 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

    ab249127 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-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    Western blot - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)

    This data was developed using ab154821, the same antibody clone in a different buffer formulation.

    Lane 1: Wild-type HAP1 cell lysate (20 µg)

    Lane 2: TRIM56 knockout HAP1 cell lysate (20 µg)

    Lane 3: MCF7 cell lysate (20 µg)

    Lane 4: A375 cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab154821 observed at 88 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab154821 was shown to specifically react with TRIM56 when TRIM56 knockout samples were used. Wild-type and TRIM56 knockout samples were subjected to SDS-PAGE. ab154821 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) andGoat anti-Mouse IgG H&L (IRDye® 680RD)preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

  • Western blot - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    Western blot - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    All lanes : Anti-TRIM56 antibody [EPR10582] (ab154821) 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 ab154821, the same antibody clone in a different buffer formulation.

    Lanes 1-4: Merged signal (red and green). Green - ab154821 observed at 88 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab154821 Anti-TRIM56 antibody [EPR10582] 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. ab154821 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 - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    Immunocytochemistry - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    This data was developed using ab154821, the same antibody clone in a different buffer formulation.ab154821 staining TRIM56 in MCF-7 (human breast carcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody. ab7291 and ab150120 were used as counterstains for primary antibody ab154821 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain. Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
    Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)
  • Flow Cytometry - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    Flow Cytometry - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    This data was developed using ab154821, the same antibody clone in a different buffer formulation.Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling TRIM56 with purified ab154821 at 1:50 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(ab150077)(1:2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black)(ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
  • Western blot - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    Western blot - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    Anti-TRIM56 antibody [EPR10582] (ab154821) at 1/1000 dilution + MCF-7 (human breast carcinoma) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 81 kDa
    Additional bands at: 81 kDa. We are unsure as to the identity of these extra bands.



    This data was developed using ab154821, the same antibody clone in a different buffer formulation.

  • Western blot - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    Western blot - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    All lanes : Anti-TRIM56 antibody [EPR10582] (ab154821) 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



    This data was developed using ab154821, the same antibody clone in a different buffer formulation.

    Lanes 1-4: Merged signal (red and green). Green - ab154821 observed at 88 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab154821 Anti-TRIM56 antibody [EPR10582] 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. ab154821 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-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    Western blot - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    Anti-TRIM56 antibody [EPR10582] (ab154821) at 1/5000 dilution + A549 (human lung carcinoma) whole cell lysates at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 81 kDa
    Additional bands at: 81 kDa. We are unsure as to the identity of these extra bands.



    This data was developed using ab154821, the same antibody clone in a different buffer formulation.

  • Western blot - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    Western blot - Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    All lanes : Anti-TRIM56 antibody [EPR10582] (ab154821) at 1/1000 dilution (unpurified)

    Lane 1 : MCF 7 cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : A375 cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 81 kDa



    This data was developed using ab154821, the same antibody clone in a different buffer formulation.

     

  • Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)
    Anti-TRIM56 antibody [EPR10582] - BSA and Azide free (ab249127)

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