Catalog # MK6370


Myosin IIA Heavy Chain Phospho-Regulation

Antibody Sampler Kit

$395

Non-muscle myosin II is an actin-based motor protein essential to cell motility, division, migration, adhesion and polarity. This myosin forms a hexameric complex comprised of two heavy chains (NMHC-II), two essential light chains, and two regulatory light chains (RLC). In vertebrates, there are three NMHC-II isoforms (NMHC-IIA, NMHC-IIB, and NMHC-IIC), which exhibit distinct patterns of expression in cells and tissues. Regulation of NMHC-II activity occurs through RLC and HC phosphorylation. RLCs are phosphorylated at Thr-18 and Ser-19, which activates myosin II motor activity and promotes filament stability. By contrast, PKC phosphorylation of Ser-1/Ser-2 and Thr-9 in RLC may decrease activated myosin II interaction with actin. Several kinases phosphorylation NMHC-II isoforms directly. TRPM7 phosphorylates Thr-1800, Ser-1803, and Ser-1808, which reduces NMHC-IIA incorporation into the actin cytoskeleton. PKC phosphorylates NMHC-IIA on Ser-1916 and NMHC-IIB on multiple tailpiece serines leading to inhibition of filament assembly. Casein kinase II phosphorylates NMHC-IIA on Ser-1943 and increases disassembly of NMHC-IIA filaments.

References
Dulyaninova, N.G. et al. (2007) Mol Biol Cell 18:3144.
Conti, M.A. & Adelstein, R.S. (2008) J Cell Sci. 121:11.
Breckenridge, M.T. et al. (2009) Mol Biol Cell 20:338.
Dulyaninova, N. & Bresnick, A. (2013) BioArchitect. 3(4):1.
Rabbit polyclonal antibodies are supplied in phosphate-buffered saline (PBS), 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.



*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

MP3831 Guilbert, A. et al. (2013) Eur J Cancer. 49(17):3694. WB: MDA-MB-231 cells
MP3831 Sanborn, K.B. et al. (2013) Blood 118(22):5862. WB: human YTS cells
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot

Western blot image of human A431 cells stimulated with calyculin A (100 nM, 30 min). The blots were untreated (lanes 1, 3 & 5) or treated with lambda phosphatase (lanes 2, 4 & 6), and probed with rabbit polyclonals Myosin IIA Heavy Chain (a.a. 1936-1950) (lanes 1 & 2), Myosin IIA Heavy Chain (Ser-1803), phospho-specific (lanes 3 & 4) or Myosin IIA Heavy Chain (Ser-1916) phospho-specific (lanes 5 & 6).

Immunocytochemical labeling of myosin IIA heavy chain phosphorylation relative to F-actin in chick fibroblasts. The cells were labeled with rabbit polyclonal Myosin IIA Heavy Chain (Ser-1943) antibody (MP3831), then detected using appropriate secondary antibody (Bottom, Red). This labeling is compared to F-actin staining (Bottom, Green) and to secondary only (Top). (Image provided by Dr. Gianluca Gallo at Drexel University).



This kit contains:

CATALOG#DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)
Myosin IIA Heavy Chain (a.a. 1936-1950) Rabbit pAb50 μlWB, E, ICCHu, Rt, Ms200
Myosin IIA Heavy Chain (Ser-1803), phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms, Ck200
Myosin IIA Heavy Chain (Ser-1916), phospho-specific Rabbit pAb 50 μlWB, EHu, Rt, Ms, Ck200
Myosin IIA Heavy Chain (Ser-1943), phospho-specific Rabbit pAb 50 μlWB, E, ICCHu, Rt, Ms, Ck200
Anti-Rabbit Ig Light Chain Specific:HRP Mouse mAb100 μlWB, E, ICC, IHCRb
KIT SUMMARY

The non-muscle myosin IIA heavy chain (NMHC-IIA) phospho-regulation antibody sampler kit can be used to detect the level of phosphorylation of NMHC-IIA at Ser-1803, Ser-1916, and Ser-1943 relative to the total expression level of NMHC-IIA. The kit includes rabbit polyclonal phospho-specific antibodies to Ser-1803, Ser-1916, and Ser-1943, a rabbit polyclonal antibody to detect total NMHC-IIA, as well as a secondary regent for detection of antibodies in western blot, ELISA, or immunocytochemistry.