Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-MGMT antibody [EPR4397] - BSA and Azide free (ab233012)

Overview

  • Product name
    Anti-MGMT antibody [EPR4397] - BSA and Azide free
    See all MGMT primary antibodies
  • Description
    Rabbit monoclonal [EPR4397] to MGMT - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human MGMT aa 1-100. The exact sequence is proprietary.
    Database link: P16455

  • Positive control
    • WB: MCF7, Jurkat, HeLa, and HT29 cell lysates IHC-P: Human clear cell carcinoma, lung cancer, and spleen tissues
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®). 

    ab233012 is a PBS-only buffer format of ab108630. Please refer to ab108630 for recommended dilutions, protocols, and image data.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab233012 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 Use at an assay dependent concentration. Detects a band of approximately 23 kDa (predicted molecular weight: 22 kDa).
IHC-P Use at an assay dependent concentration.

Antigen retrieval is recommended. See IHC antigen retrieval protocols.

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

  • 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 - unpurified ab108630 observed at 22 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab108630 was shown to specifically react with MGMT when MGMT knockout samples were used. Wild-type and MGMT knockout samples were subjected to SDS-PAGE. ab108630 and ab8245 (loading control to GAPDH) 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108630).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung cancer tissue sections labeling MGMT with purified ab108630 at 1:800 dilution (0.94 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using Citrate buffer, pH 6.0. 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�(ab108630)
  • Unpurified ab108630, at a 1/250 dilution, staining MGMT in paraffin embedded Human clear cell carcinoma tissue by Immunohistochemistry.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108630).

  • Unpurified ab108630, at a 1/250 dilution, staining MGMT in paraffin embedded Human spleen tissue by Immunohistochemistry.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108630).

References

ab233012 has not yet been referenced specifically in any publications.

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