Product nameAnti-MGMT antibody [MT3.1]
See all MGMT primary antibodies
DescriptionMouse monoclonal [MT3.1] to MGMT
Tested applicationsSuitable for: IHC-Fr, WB, IHC-FoFr, ICC, Flow Cyt, ICC/IF, IHC-Pmore details
Species reactivityReacts with: Human
Recombinant full length protein corresponding to Human MGMT.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.05% Sodium azide
Constituents: 1% BSA, Tissue culture supernatant
Concentration information loading...
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.
|WB||1/500. Detects a band of approximately 21 kDa.|
|IHC-FoFr||1/25 - 1/50.|
|ICC||Use at an assay dependent concentration.|
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.|
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.
- Information by UniProt
- 6 O methylguanine DNA methyltransferase antibody
- 6-O-methylguanine-DNA methyltransferase antibody
- Agat antibody
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) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (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.
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.
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.
All lanes : Anti-MGMT antibody [MT3.1] (ab39253) at 1/500 dilution (for 16 hours at 4°C)
All lanes : Human OE33 cells - whole cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : 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
Blocking Step: 5% BSA for 1 hour at 23°C
Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human tonsil tissue, staining MGMT with ab39253.
This product has been referenced in:
- Chen TC et al. Rare Stochastic Expression of O6-Methylguanine- DNA Methyltransferase (MGMT) in MGMT-Negative Melanoma Cells Determines Immediate Emergence of Drug-Resistant Populations upon Treatment with Temozolomide In Vitro and In Vivo. Cancers (Basel) 10:N/A (2018). Read more (PubMed: 30274152) »
- Zink LM et al. H3.Y discriminates between HIRA and DAXX chaperone complexes and reveals unexpected insights into human DAXX-H3.3-H4 binding and deposition requirements. Nucleic Acids Res 45:5691-5706 (2017). Read more (PubMed: 28334823) »