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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.|
|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.|
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.
This image is courtesy of an anonymous AbreviewBlocking Step: 5% BSA for 1 hour at 23°C