Overview

  • Product nameAnti-MGMT antibody [MT3.1]
    See all MGMT primary antibodies
  • Description
    Mouse monoclonal [MT3.1] to MGMT
  • Tested applicationsSuitable for: IHC-Fr, WB, IHC-FoFr, ICC, Flow Cyt, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein corresponding to Human MGMT.

  • Positive control
    • Tonsil

Properties

Applications

Our Abpromise guarantee covers the use of ab39253 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
IHC-Fr 1/50.
WB 1/500. Detects a band of approximately 21 kDa.
IHC-FoFr 1/25 - 1/50.
ICC Use at an assay dependent concentration.
Flow Cyt 1/50. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF Use at an assay dependent concentration.
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • FunctionInvolved 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 similaritiesBelongs to the MGMT family.
  • Cellular localizationNucleus.
  • 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

Anti-MGMT antibody [MT3.1] 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 - ab39253 observed at 22 kDa. Red - loading control, ab181602, observed at 37 kDa.
    ab39253 was shown to specifically react with MGMT when MGMT knockout samples were used. Wild-type and MGMT knockout samples were subjected to SDS-PAGE. ab39253 and ab181602 (loading control to GAPDH) were diluted 1/500 and 1/10 000 and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) ab216772 and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) ab216777 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • ab39253 staining MGMT in Human stomach tissue sections by IHC-Fr (frozen sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at 23°C. Samples were incubated with primary antibody (1/50 in blocking buffer) for 1 hour at 23°C. An undiluted HRP-conjugated goat anti-mouse IgG polyclonal was used as secondary antibody.

    See Abreview

  • ab39253 staining MGMT in human stomach tissue sections by Immunohistochemistry. Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using 10mM citrate buffer pH6.0. Samples were then blocked with 5% serum for 1 hour at 23°C followed by incubation with the primary antibody, at a 1/100 dilution, for 1 hour at 23°C. An undiluted HRP-conjugated goat anti-mouse IgG polyclonal was used as secondary antibody.

    See Abreview

  • All lanes : Anti-MGMT antibody [MT3.1] (ab39253) at 1/500 dilution (for 16 hours at 4°C)

    Lane 1 : Human OE33 cells - whole cell lysate
    Lane 2 : Human OE33 cells - whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/1000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 21 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

    This image is courtesy of an anonymous Abreview

    Blocking Step: 5% BSA for 1 hour at 23°C

    See Abreview

  • Overlay histogram showing Jurkat cells stained with ab39253 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab39253, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human tonsil tissue, staining MGMT with ab39253.

References for Anti-MGMT antibody [MT3.1] (ab39253)

This product has been referenced in:
  • Viel T  et al. Optimizing Glioblastoma Temozolomide Chemotherapy Employing Lentiviral-based Anti-MGMT shRNA Technology. Mol Ther 21:570-9 (2013). Read more (PubMed: 23319055) »
  • Viel T  et al. Early assessment of the efficacy of temozolomide chemotherapy in experimental glioblastoma using [18F]FLT-PET imaging. PLoS One 8:e67911 (2013). WB ; Human . Read more (PubMed: 23861829) »

See all 4 Publications for this product

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We do not have expiration dates for our products, but we do guarantee our products for 6 months if stored according to the datasheet. Although we cannot guarantee it, based on our in-house testing if the antibo...

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Thank you for your reply. The publications that are known to reference an Abcam product can be found on our online datasheets on the specific references tab. Below are PubMed IDs for the antibodies you are interested in.

MGMT (ab39253) - 2175...

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Thank you for contacting us.

The final dilutions should be optimized for your particular samples and protocols, but the following would be my recommended starting dilutions for WB with an overnight incubation:

MGMT (ab39253) - 1:500...

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I can confirm that theaverage concentration of this antibody is 66.6 ug/ml.


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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - other (OE33 cells)
Loading amount 20 µg
Specification OE33 cells
Gel Running Conditions Reduced Denaturing
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Aug 27 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (AGS Gastric Carcinoma cells)
Specification AGS Gastric Carcinoma cells
Fixative Formaldehyde
Permeabilization Yes - 0.025% Triton-X in TBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Jun 02 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry
Sample Human Cultured Cells (AGS Gastric Carcinoma cells)
Specification AGS Gastric Carcinoma cells
Fixative Formaldehyde
Permeabilization Yes - 0.025% Triton-X in TBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted May 19 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Stomach)
Specification Stomach
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10mM citrate pH6.0
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Mar 24 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (Stomach)
Specification Stomach
Fixative Paraformaldehyde
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Mar 18 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"