The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
1/500 - 1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
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.
All lanes : Anti-MGMT antibody (ab69629) at 1/500 dilution
Lane 1 : Extracts from Jurkat cells. Lane 2 : Extracts from Jurkat cells with immunising peptide at 5 µg
Lysates/proteins at 5 µg per lane.
Predicted band size: 22 kDa Observed band size: 22 kDa
Immunocytochemistry/ Immunofluorescence - MGMT antibody (ab69629)This image is courtesy of an Abreview submitted by Mark Willett.
ab69629 staining MGMT in T98G human glioma cell line by Immunocytochemistry/ Immunofluorescence. Cells were fixed in paraformaldehyde and permeabilized in 0.1% Triton X-100 prior to blocking in 3% BSA for 30 minutes at 20°C. The primary antibody was diluted 1/100 and incubated with the sample for 1 hour at 20°C. The secondary antibody was Alexa Fluor® 555-conjugated goat anti-rabbit polyclonal, diluted 1/300. MGMT staining is shown in red (AF 555) and stains nuclei and to a small extent, cytoplasm. Nuclei were stained with DAPI (blue), while microtubules were stained using an anti-tubulin FITC conjugate (green).
Moen EL et al. Characterization of CpG sites that escape methylation on the inactive human X-chromosome. Epigenetics10:810-8 (2015).
Read more (PubMed: 26178744) »
Ishiguro K et al. Expression of O(6)-Methylguanine-DNA Methyltransferase Examined by Alkyl-Transfer Assays, Methylation-Specific PCR and Western Blots in Tumors and Matched Normal Tissue. J Cancer Ther4:919-931 (2013).
Read more (PubMed: 23946891) »