Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22825-2] to TRIM24 - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free
    See all TRIM24 primary antibodies

  • Description

    Rabbit monoclonal [EPR22825-2] to TRIM24 - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    This antibody is not recommended for mouse IHC.

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type HAP1 whole, HCT116, HeLa, A549, PC-3, WEHI-3, P815, CTLL-2 and C2C12 whole cell lysate. Mouse colon lysates. IHC-P: Human lung cancer and breast cancer tissue. ICC/IF: HepG2 and wild type HAP1 cells.

  • General notes

    ab260000 is the carrier-free version of ab256491.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)
    Western blot - Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)
    All lanes : Anti-TRIM24 antibody [EPR22825-2] (ab256491) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : TRIM24 knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 116 kDa
    Observed band size: 140 kDa
    why is the actual band size different from the predicted?



    The expression profile observed is consistent with what has been described in the literature (PMID: 19909775).

    ab256491 was shown to specifically react with TRIM24 in wild-type HAP1 cells as signal was lost in TRIM24 knockout cells. Wild-type and TRIM24 knockout samples were subjected to SDS-PAGE. ab256491 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique. Blocking/Diluting buffer and concentration: 5% NFDM/TBST. Exposure Time: 48 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256491).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)

    Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling TRIM24 with ab256491 at 1/500 dilution (1.08µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in human breast cancer (PMID: 21164480) is observed. The section was incubated with ab255611 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256491).

  • Western blot - Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)
    Western blot - Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)
    All lanes : Anti-TRIM24 antibody [EPR22825-2] (ab256491) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : TRIM24 knockout HeLa cell lysate
    Lane 3 : Hap1 cell lysate
    Lane 4 : A549 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 116 kDa
    Observed band size: 140 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 (ab256491).

      Lanes 1- 2: Merged signal (red and green). Green - ab256491 observed at 140 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab256491 was shown to react with Tripartite Motif Containing 24 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264963 (knockout cell lysate ab258246) lane below 140kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and TRIM24 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab256491 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 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-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)

    Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling TRIM24 with ab256491 at 1/500 dilution (1.08µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in human lung cancer (PMID: 22666376) is observed. The section was incubated with ab255611 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256491).

  • Immunocytochemistry/ Immunofluorescence - Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)
    Immunocytochemistry/ Immunofluorescence - Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Wild type HAP1 (Wild type human chronic myelogenous leukemia near-haploid cell line) and TRIM24 knockout HAP1 (TRIM24 knockout human chronic myelogenous leukemia near-haploid cell line) cells labeling TRIM24 with ab256491 at 1/100 5.4 µg/ml dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 2 µg/ml dilution (Green). Confocal image showing nuclear staining in Wild type HAP1 cell line, and no staining in TRIM24 knockout HAP1 cell line. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 2 µg/ml dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256491).

  • Immunocytochemistry/ Immunofluorescence - Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)
    Immunocytochemistry/ Immunofluorescence - Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labeling TRIM24 with ab256491 at 1/100 5.4 µg/ml dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 2 µg/ml dilution (Green). Confocal image showing nuclear staining in HepG2 cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 2 µg/ml dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256491).

  • Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)
    Anti-TRIM24 antibody [EPR22825-2] - BSA and Azide free (ab260000)

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