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Our Abpromise guarantee covers the use of ab125114 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||1/50. Fixed 2 hr in 2% paraformaldehyde and 0.1 % glutaraldehyde in 0.1 M PB.|
|ICC/IF||Use at an assay dependent concentration.|
|WB||1/50. Detects a band of approximately 43-46 kDa (predicted molecular weight: 43 kDa).|
|IHC-P||1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. 0.5 % pepsin in 0.01 N HCl for 15 min at 37 °C.|
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Creatine kinase B knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: SH-SY5Y cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab125114 observed at 45 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab125114 was shown to recognize Creatine kinase B when Creatine kinase B knockout samples were used, along with additional cross-reactive bands. Wild-type and Creatine kinase B knockout samples were subjected to SDS-PAGE. ab125114 and ab181602 (loading control to GAPDH) were diluted 1/50 and 1/10000 respectively 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 1/10000 dilution for 1 hour at room temperature before imaging.