ab71960 was purified using protein G column chromatography from culture supernatant of hybridoma cultured in a medium containing bovine IgG-depleted (approximately 95%) fetal bovine serum and filtered through a 0.22µm membrane.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. Detects a band of approximately 37 kDa (predicted molecular weight: 56 kDa).
Stress-activated, pro-apoptotic kinase which, following caspase-cleavage, enters the nucleus and induces chromatin condensation followed by internucleosomal DNA fragmentation. Key component of the Hippo signaling pathway which plays a pivotal role in organ size control and tumor suppression by restricting proliferation and promoting apoptosis. The core of this pathway is composed of a kinase cascade wherein MST1/MST2, in complex with its regulatory protein SAV1, phosphorylates and activates LATS1/2 in complex with its regulatory protein MOB1, which in turn phosphorylates and inactivates YAP1 oncoprotein and WWTR1/TAZ. Phosphorylation of YAP1 by LATS2 inhibits its translocation into the nucleus to regulate cellular genes important for cell proliferation, cell death, and cell migration. MST1/MST2 are required to repress proliferation of mature hepatocytes, to prevent activation of facultative adult liver stem cells (oval cells), and to inhibit tumor formation. Phosphorylates NKX2-1.
Expressed at high levels in adult kidney, skeletal and placenta tissues and at very low levels in adult heart, lung and brain tissues.
Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. STE20 subfamily. Contains 1 protein kinase domain. Contains 1 SARAH domain.
Cytoplasm. Nucleus. The caspase-cleaved form cycles between nucleus and cytoplasm.
Western blot - Anti-STK3 antibody [3067C3a] (ab71960)
Predicted band size : 56 kDa
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: STK3 knockout HAP1 cell lysate (20 µg) Lane 3: NIH/3T3 cell lysate (20 µg) Lane 4: 293T cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab71960 observed at 56 kDa. Red - loading control, ab181602, observed at 37 kDa. ab71960 was shown to recognize STK3 when STK3 knockout samples were used, along with additional cross-reactive bands. Wild-type and STK3 knockout samples were subjected to SDS-PAGE. ab71960 and ab181602 (loading control to STK3) were diluted 1 μg/mL and 1/10 000 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 1/10 000 dilution for 1 h at room temperature before imaging.
Western blot - STK3 antibody [3067C3a] (ab71960)
Anti-STK3 antibody [3067C3a] (ab71960) + immunising recombinant protein
Predicted band size : 56 kDa Observed band size : 37 kDa (why is the actual band size different from the predicted?) The molecular weight of the band on the western blot does not correspond to the molecular weight of the natural protein because only a fragment of the protein was used.