The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500 - 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
1/5000 - 1/15000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
Use at 2-5 µg/mg of lysate.
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human placenta tissue labelling STK3 with ab70546 at 1/1000 (1µg/ml). Detection: DAB.
Western blot - STK3 antibody (ab70546)
All lanes : Anti-STK3 antibody (ab70546) at 1/10000 dilution
Lane 1 : Whole cell lysate from HeLa cells at 50 µg Lane 2 : Whole cell lysate from HeLa cells at 15 µg Lane 3 : Whole cell lysate from HeLa cells at 5 µg Lane 4 : Whole cell lysate from Ramos cells at 50 µg Lane 5 : Whole cell lysate from NIH3T3 cells at 50 µg
Developed using the ECL technique.
Predicted band size: 56 kDa Observed band size: 56 kDa
Exposure time: 3 seconds
Immunoprecipitation - STK3 antibody (ab70546)
1mg whole cell lysate from HeLa cells was immunoprecipitated with ab70546 at 3ug/mg of lysate (lane 1) or a control rabbit Ig )lane 2). For the subsequent blot, 25% of the immnuoprecipitate (lanes 1 and 2) or the input (lane 3) were probed with ab70546 at 1mg/ml.