An antibody for phosphorylated myosin light chain of smooth muscle: application to a biochemical study.
An antibody with specificity for the 20 kDa myosin light chain of smooth muscle phosphorylated by myosin light chain kinase was developed. The antibody was raised against the phosphorylated synthetic peptide, Lys-Lys-Arg-Pro-Gln-Arg-Ala-Thr-phospho-Ser-Asn-Val-Phe-Cys (residues 11-22 of the myosin light chain). This antibody only recognized the monophosphorylated myosin light chain at serine 19, i.e., with no detectable recognition of nonphosphorylated or diphosphorylated serine 19 or threonine 18. The specificity was tested by EIA assaying of myosin light chain kinase activity using a 96-well plate coated with the light chain. This EIA system was as sensitive as the radioisotopic method, and the newly developed non-radioisotopic method.
Sakurada
K
K
Life Science Center, Asahi Chem. Indust. Co., Ltd., Miyazaki.
Ikuhara
T
T
Seto
M
M
Sasaki
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009218:Myosins / Q000276:immunology / Q000378:metabolism*; D010766:Phosphorylation; D011817:Rabbits
D009219:Myosin-Light-Chain Kinase; D009218:Myosins