Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4397] to MGMT
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Product nameAnti-MGMT antibody [EPR4397]
See all MGMT primary antibodies
DescriptionRabbit monoclonal [EPR4397] to MGMT
Tested applicationsSuitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt,ICC or IP
Species reactivityReacts with: Human
Synthetic peptide within Human MGMT aa 1-100. The exact sequence is proprietary.
Database link: P16455
- WB: MCF7, Jurkat, HeLa, and HT29 cell lysates IHC-P: Human clear cell carcinoma, lung cancer, and spleen tissues
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
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.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab108630 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Detects a band of approximately 23 kDa (predicted molecular weight: 22 kDa).|
|IHC-P||1/800. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Antigen retrieval is recommended. See IHC antigen retrieval protocols.
For unpurified use at 1/250- 1/500.
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
Anti-MGMT antibody [EPR4397] (ab108630) at 1/5000 dilution (Purified) + MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 22 kDa
Observed band size: 22 kDa
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.
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 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.
All lanes : Anti-MGMT antibody [EPR4397] (ab108630) at 1/1000 dilution (Unpurified)
Lane 1 : MCF7 cell lysate
Lane 2 : Jurkat ell lysate
Lane 3 : HeLa cell lysate
Lane 4 : HT29 cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 22 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?
Unpurified ab108630, at a 1/250 dilution, staining MGMT in paraffin embedded Human clear cell carcinoma tissue by Immunohistochemistry.
Unpurified ab108630, at a 1/250 dilution, staining MGMT in paraffin embedded Human spleen tissue by Immunohistochemistry.
ab108630 has been referenced in 5 publications.
- von Roemeling CA et al. Therapeutic modulation of phagocytosis in glioblastoma can activate both innate and adaptive antitumour immunity. Nat Commun 11:1508 (2020). PubMed: 32198351
- Lai SW et al. Differential Characterization of Temozolomide-Resistant Human Glioma Cells. Int J Mol Sci 19:N/A (2018). PubMed: 29301329
- Isohookana J et al. Proteins of the retinoblastoma pathway, FEN1 and MGMT are novel potential prognostic biomarkers in pancreatic adenocarcinoma. Pathol Res Pract 214:840-847 (2018). IHC-P ; Human . PubMed: 29735403
- Wang X et al. Coaxial extrusion bioprinted shell-core hydrogel microfibers mimic glioma microenvironment and enhance the drug resistance of cancer cells. Colloids Surf B Biointerfaces 171:291-299 (2018). PubMed: 30048904
- Niittykoski M et al. Immunohistochemical Characterization and Sensitivity to Human Adenovirus Serotypes 3, 5, and 11p of New Cell Lines Derived from Human Diffuse Grade II to IV Gliomas. Transl Oncol 10:772-779 (2017). PubMed: 28797937