Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-MGMT antibody [EPR4398] (ab108989)

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Overview

  • Product name

    Anti-MGMT antibody [EPR4398]
    See all MGMT primary antibodies

  • Description

    Rabbit monoclonal [EPR4398] to MGMT

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: ICC/IF or IHC-P

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human MGMT aa 150 to the C-terminus.

  • Positive control

    • MCF7 and Jurkat cell lysates

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these 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.

    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

Applications

Our Abpromise guarantee covers the use of ab108989 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/10000. Detects a band of approximately 23 kDa (predicted molecular weight: 22 kDa).
IP 1/10 - 1/100.
  • Application notes
    Is unsuitable for ICC/IF or IHC-P.

    • Target

    • Function

      Involved in the cellular defense against the biological effects of O6-methylguanine (O6-MeG) in DNA. Repairs alkylated guanine in DNA by stoichiometrically transferring the alkyl group at the O-6 position to a cysteine residue in the enzyme. This is a suicide reaction: the enzyme is irreversibly inactivated.

    • Sequence similarities

      Belongs to the MGMT family.

    • Cellular localization

      Nucleus.
    • Information by UniProt

    • Database links

      • Alternative names

        • 6 O methylguanine DNA methyltransferase antibody
        • 6-O-methylguanine-DNA methyltransferase antibody
        • Agat antibody
        • AGT antibody
        • AI267024 antibody
        • EC 2.1.1.63 antibody
        • Methylated DNA protein cysteine methyltransferase antibody
        • Methylated-DNA--protein-cysteine methyltransferase antibody
        • Methylguanine DNA methyltransferase antibody
        • MGC107020 antibody
        • MGMT antibody
        • MGMT_HUMAN antibody
        • O 6 methylguanine DNA alkyltransferase antibody
        • O 6 methylguanine DNA methyltransferase antibody
        • O-6-methylguanine-DNA methyltransferase antibody
        • O-6-methylguanine-DNA-alkyltransferase antibody
        see all

      Images

      • Western blot - Anti-MGMT antibody [EPR4398] (ab108989)
        Western blot - Anti-MGMT antibody [EPR4398] (ab108989)

        Lane 1: Wild-type HAP1 cell lysate (20 µg)
        Lane 2: MGMT knockout HAP1 cell lysate (20 µg)
        Lane 3: MCF7 cell lysate (20 µg)
        Lane 4: Jurkat cell lysate (20 µg)
        Lanes 1 - 4: Merged signal (red and green). Green - ab108989 observed at 22 kDa. Red - loading control, ab8245, observed at 37 kDa.
        ab108989 was shown to specifically react with MGMT when MGMT knockout samples were used. Wild-type and MGMT knockout samples were subjected to SDS-PAGE. ab108989 and ab8245 (loading control to MGMT) were diluted 1/1000 and 1/2000 respectively and 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 h at room temperature before imaging.

      • Western blot - Anti-MGMT antibody [EPR4398] (ab108989)
        Western blot - Anti-MGMT antibody [EPR4398] (ab108989)
        All lanes : Anti-MGMT antibody [EPR4398] (ab108989) at 1/1000 dilution (purified)

        Lane 1 : MCF7 cell lysate
        Lane 2 : Jurkat cell lysate

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

        Predicted band size: 22 kDa
        Observed band size: 23 kDa
        why is the actual band size different from the predicted?



        Blocking buffer: 5% NFDM/TBST
        Dilution buffer: 5% NFDM/TBST

      • Immunoprecipitation - Anti-MGMT antibody [EPR4398] (ab108989)
        Immunoprecipitation - Anti-MGMT antibody [EPR4398] (ab108989)

        ab108989 (purified) at 1/30 immunoprecipitating MGMT in 10 μg MCF7 (Lanes 1 and 2, observed at 23 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

      • Western blot - Anti-MGMT antibody [EPR4398] (ab108989)
        Western blot - Anti-MGMT antibody [EPR4398] (ab108989)
        All lanes : Anti-MGMT antibody [EPR4398] (ab108989) at 1/1000 dilution (unpurified)

        Lane 1 : MCF7 cell lysate
        Lane 2 : Jurkat cell lysate

        Lysates/proteins at 10 µg per lane.

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

        Predicted band size: 22 kDa
        Observed band size: 23 kDa why is the actual band size different from the predicted?

      References

      ab108989 has not yet been referenced specifically in any publications.

      Publishing research using ab108989? Please let us know so that we can cite the reference in this datasheet.

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      Question

      Many thanks for your reply. The answer was perfect.
      I was also wondering whether you have any information regarding the concentration of the particular Lot that I have?

      Read More
      Answer

      Thank you for your reply.

      ab108989 is atissue culture supernatant, not a purified antibody.
      Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet.

      The concentration estimate of antibodyin tissue culture supernatants is usuallyin the range of1 - 3 mg/ml.

      I am sorry I cannot provide an exact concentration on this occasion, but I hope this information is nevertheless helpful to you.
      If you have any further questions, please do not hesitate to contact us.

      Read More

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