1/10 - 1/100. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Reversibly catalyzes the transfer of phosphate between ATP and various phosphogens (e.g. creatine phosphate). Creatine kinase isoenzymes play a central role in energy transduction in tissues with large, fluctuating energy demands, such as skeletal muscle, heart, brain and spermatozoa.
Belongs to the ATP:guanido phosphotransferase family. Contains 1 phosphagen kinase C-terminal domain. Contains 1 phosphagen kinase N-terminal domain.
Western blot - Anti-Creatine kinase B type antibody [EPR3926] (ab108388)
Predicted band size : 43 kDa
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 - ab108388 observed at 45 kDa. Red - loading control, ab18058, observed at 124 kDa. ab108388 was shown to specifically react with Creatine kinase B when Creatine kinase B knockout samples were used. Wild-type and Creatine kinase B knockout samples were subjected to SDS-PAGE. ab108388 and ab18058 (loading control to vinculin) were diluted 1/50000 and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.
Flow Cytometry - Anti-Creatine kinase B type antibody [EPR3926] (ab108388)
Overlay histogram showing SH-SY5Y cells stained with ab108388 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab108388, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Western blot - Creatine Kinase BB antibody [EPR3926] (ab108388)
All lanes : Anti-Creatine kinase B type antibody [EPR3926] (ab108388) at 1/50000 dilution
Lane 1 : 293T cell lysate Lane 2 : SH SY5Y cell lysate Lane 3 : Y79 cell lysate Lane 4 : HeLa cell lysate