K-K-R-P-Q-R-A-T-(pS)-N-V-F-A-M-F-D (aa 12-27 of Human sequence, aa 12-28 of Mouse)
Specificity
This antibody is specific for the phosphorylated form of myosin light chain. This affinity purified antibody is directed against the regulatory light chain of smooth and nonmuscle myosin. The antibody is phosphospecific and detect monophosphorylated and diphosphorylated forms of the protein. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide. This phospho specific polyclonal antibody is specific for phosphorylated pS20 of the protein. Reactivity with non-phosphorylated myosin light chain is less than 1 % by ELISA. Cross reactivity with myosin light chain from other species has not been determined, however, the sequence of the immunogen is identical in mammalian and avian species. BLAST search analysis was used to determine that the smooth and non-muscle forms of myosin regulatory light chain have identical sequences. Cross reactivity is expected.
Cross-Reactivity (Details)
Species reactivity (tested):Human.
Purification
Immunoaffinity Chromatography.
Immunogen
Human Myosin Light Chain phospho peptide corresponding to a region of the Human smooth/non-muscle form of Myosin regulatory light chain conjugated to Keyhole Limpet Hemocyanin (KLH).
MYL12A
Reactivity: Human
WB
Host: Rabbit
Polyclonal
unconjugated
Application Notes
This phospho specific polyclonal antibody was tested by ELISA, Immunohistochemistryand Immunoblotting and was found to be reactive with unstimulated and stimulatedcardiac myocytes and the phosphorylated form of the immunizing peptide and minimallyreactive with the non-phosphorylated form of the immunizing peptide.
Restrictions
For Research Use only
Concentration
0.92 mg/mL (by UV absorbance at 280 nm)
Buffer
0.02 M Potassium Phosphate, 0.12 M Sodium Chloride, pH 7.2 with 0.01 % (w/v) Sodium Azide as preservative.
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
4 °C
Storage Comment
Store undiluted at 2-8 °C. DO NOT FREEZE!
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Lai, Martinka, Fähling, Mrowka, Steege, Gericke, Sendeski, Persson, Persson, Patzak: "Adenosine restores angiotensin II-induced contractions by receptor-independent enhancement of calcium sensitivity in renal arterioles." in: Circulation research, Vol. 99, Issue 10, pp. 1117-24, (2006) (PubMed).
Myosin is the major component of thick muscle filaments, and is a long asymmetric molecule containing a globular head and a long tail. The molecule consists of two heavy chains each ~200,000 daltons, and four light chains each ~16,000 - 21,000 daltons. Activation of smooth and cardiac muscle primarily involves pathways which increase calcium and myosin phosphorylation resulting in contraction. Myosin light chain phosphatase acts to regulate muscle contraction by dephosphorylating activated myosin light chain. The selected peptide sequence used to generate the polyclonal antibody is located near the amino terminal end of the polypeptide corresponding to the smooth/non-muscle form of myosin regulatory light chain found in cardiac myocytes in addition to smooth and non-muscle cells. This sequence differs from that of the sarcomeric/cardiac form of myosin regulatory light chain that has a different sequence around the phosphorylation site. Human and mouse have almost identical sequences. In human the phosphorylation site is pS19, while in mouse the site maps to pS20.Synonyms: MLC-2B, MLCB, MRLC3, Myosin RLC, Myosin regulatory light chain 12A, Myosin regulatory light chain 2 nonsarcomeric, Myosin regulatory light chain MRLC3, RLC