The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesWB: 1/250. Detects a band of approximately 52 kDa (predicted molecular weight: 50 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
FunctionImplicated in the hormonal control of several regulatory proteins including glycogen synthase, MYB and the transcription factor JUN.
Sequence similaritiesBelongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. GSK-3 subfamily. Contains 1 protein kinase domain.
Lane 1: GSK3A knockout HAP1 whole cell lysate (20 µg) Lane 2: Wild type HAP1 whole cell lysate (20 µg) Lane 3: HeLa whole cell lysate (20 µg) Lane 4: HepG2 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab78664 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab78664 was shown to recognize GSK3A when GSK3A knockout samples were used, along with additional cross-reactive bands. Wild-type and GSK3A knockout samples were subjected to SDS-PAGE. Ab78664 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 250 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - GSK3 alpha antibody (ab78664)
All lanes : Anti-GSK3 alpha antibody (ab78664) at 1/250 dilution
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution Developed using the ECL technique