1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Serine/threonine kinase that plays a key role in M phase by acting as a regulator of mitosis entry and maintenance. Acts by promoting the inactivation of protein phosphatase 2A (PP2A) during M phase: does not directly inhibit PP2A but acts by mediating phosphorylation and subsequent activation of ARPP19 and ENSA at 'Ser-62' and 'Ser-67', respectively. ARPP19 and ENSA are phosphatase inhibitors that specifically inhibit the PPP2R2D (PR55-delta) subunit of PP2A. Inactivation of PP2A during M phase is essential to keep cyclin-B1-CDK1 activity high. Following DNA damage, it is also involved in checkpoint recovery by being inhibited. Phosphorylates histone protein in vitro; however such activity is unsure in vivo. May be involved in megakaryocyte differentiation.
Involvement in disease
Defects in MASTL are the cause of thrombocytopenia type 2 (THC2) [MIM:188000]. Thrombocytopenia is defined by a decrease in the number of platelets in circulating blood, resulting in the potential for increased bleeding and decreased ability for clotting.
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. Contains 1 AGC-kinase C-terminal domain. Contains 1 protein kinase domain.
Phosphorylation at Thr-741 by CDK1 during M phase activates its kinase activity (By similarity). Maximum phosphorylation occurs in prometaphase.
Cytoplasm > cytoskeleton > centrosome. Nucleus. Cleavage furrow. During interphase is mainly nuclear, upon nuclear envelope breakdown localizes at the cytoplasm and during mitosis at the centrosomes. Upon mitotic exit moves to the cleavage furrow.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testicular seminoma (left) and lung carcinoma (right) tissues labelling MASTL with ab86387 at 1/1000 (1µg/ml) and 1/500 (2µg/ml). Detection: DAB and a DyLight® 594-conjugated goat anti-rabbit IgG (H+L) (1/100).
Western blot - Anti-MASTL antibody (ab86387)
All lanes : Anti-MASTL antibody (ab86387) at 0.1 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg Lane 4 : 293T whole cell lysate at 50 µg
Detection of MASTL by Western blot of Immunprecipitate.
ab86387, at 1 µg/ml, staining MASTL in HeLa whole cell lysate immunoprecipitated using ab86387 at 3 µg/mg lysate (1 mg/IP; 20% of IP loaded/lane). Detection: Chemiluminescence with an exposure time of 10 seconds.