Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR15934-50] to MTH1 - BSA and Azide free
  • Suitable for: WB, IHC-P, IP, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free
    See all MTH1 primary antibodies

  • Description

    Rabbit monoclonal [EPR15934-50] to MTH1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

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

  • Immunogen

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

  • Positive control

    • WB: HEK-293T, HAP1, HeLa, Jurkat and A549 cell lysates. IHC-P: Human thymus and squamous cell carcinoma of lung tissues. ICC/IF: Jurkat and A549 cells. Flow Cyt: Jurkat cells. IP: Jurkat whole cell lysate.

  • General notes

    Ab246327 is the carrier-free version of ab200832. 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

    ab246327 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)
    Western blot - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)
    All lanes : Anti-MTH1 antibody [EPR15934-50] (ab200832) at 1/5000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : NUDT1 knockout HEK-293T cell lysate
    Lane 3 : HAP1 cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 23 kDa
    Observed band size: 18 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 (ab200832).

    Lanes 1- 4: Merged signal (red and green). Green - ab200832 observed at 18 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab200832 was shown to react with MTH1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266400 (knockout cell lysate ab257565) was used. Wild-type HEK-293T and NUDT1 knockout HEK-293T 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. ab200832 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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) 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-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)
    Immunocytochemistry/ Immunofluorescence - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (Human lung carcinoma) cells labeling MTH1 with ab200832 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Cytoplasmic and nuclear staining on A549 cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:-
    -ve control 1: ab200832 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

  • Immunocytochemistry/ Immunofluorescence - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)
    Immunocytochemistry/ Immunofluorescence - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling MTH1 with ab200832 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Cytoplasmic and nuclear staining on Jurkat cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:-
    -ve control 1: ab200832 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

  • Western blot - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)
    Western blot - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: MTH1 knockout HAP1 cell lysate (20 µg)
    Lane 3: Jurkat cell lysate (20 µg)
    Lane 4: A549 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab200832 observed at 18 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab200832 was shown to specifically react with MTH1 when MTH1 knockout samples were used. Wild-type and MTH1 knockout samples were subjected to SDS-PAGE. ab200832 at a dilution of 1/5000 and ab8245 (loading control to GAPDH) diluted to 1/10,000 were incubated overnight at 4°C. 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 1/10,000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

  • Immunoprecipitation - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)
    Immunoprecipitation - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)

    MTH1 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate with ab200832 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab200832 at 1/2000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: Jurkat whole cell lysate 10ug (Input).
    Lane 2: ab200832 IP in Jurkat whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200832 in Jurkat whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

  • Flow Cytometry - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)
    Flow Cytometry - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)

    Flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling MTH1 with ab200832 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)

    Immunohistochemical analysis of paraffin-embedded Human squamous cell carcinoma of lung tissue labeling MTH1 with ab200832 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining on Human squamous cell carcinoma of lung tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)

    Immunohistochemical analysis of paraffin-embedded Human thymus tissue labeling MTH1 with ab200832 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining on Human thymus tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)
    Immunocytochemistry/ Immunofluorescence - Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)

    Immunocytochemistry/Immunofluorescence analysis of Daudi (human Burkitt's lymphoma) labelling MTH-1 with purified ab200832 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

    Control: PBS only

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200832).

  • Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)
    Anti-MTH1 antibody [EPR15934-50] - BSA and Azide free (ab246327)

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