The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Predicted molecular weight: 90 kDa.
Use at an assay dependent concentration.
1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Application notesIs unsuitable for ICC.
FunctionSerine/threonine kinase that may play a role in mediating the growth-factor and stress induced activation of the transcription factor CREB.
Tissue specificityExpressed in many tissues, highest levels in skeletal muscle.
Involvement in diseaseDefects in RPS6KA3 are the cause of Coffin-Lowry syndrome (CLS) [MIM:303600]; an X-linked dominant disorder characterized by severe mental retardation with facial and digital dysmorphisms, and progressive skeletal deformations.
Sequence similaritiesBelongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. S6 kinase subfamily. Contains 1 AGC-kinase C-terminal domain. Contains 2 protein kinase domains.
Post-translational modificationsAutophosphorylated on Ser-386, as part of the activation process. Ser-227 phosphorylation promotes Ser-386 phosphorylation and leads to basal activation. Full activation by growth factors requires additional phosphorylation on Ser-369.
Western blot - Anti-Rsk 2 / MAPKAP Kinase 1b antibody [Y82] (ab32062)
Predicted band size : 90 kDa
Lane 1: Wild-type HAP1 cell lysate (40 µg) Lane 2: Rsk 2 / MAPKAP Kinase 1b knockout HAP1 cell lysate (40 µg) Lane 3: MCF7 cell lysate (40 µg) Lane 4: HeLa cell lysate (40 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab32062 observed at 88 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32062 was shown to recognize Rsk 2 / MAPKAP Kinase 1b when Rsk 2 / MAPKAP Kinase 1b knockout samples were used, along with additional cross-reactive bands. Wild-type and Rsk 2 / MAPKAP Kinase 1b knockout samples were subjected to SDS-PAGE. Ab32062 and ab8245 (loading control to GAPDH) were diluted at 1/500 and 1/10000 dilution respectively and incubated overnight at 4C. 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.
Overlay histogram showing MCF7 cells stained with ab32062 (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 (ab32062, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 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. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Western blot - Rsk 2 / MAPKAP Kinase 1b antibody [Y82] (ab32062)