This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids aa 10-35 of human myosin light chain protein.
This affinity-purified antibody was tested by ELISA and immunoblotting and was found to be reactive with both the unphosphorylated and mono-phosphorylated forms of the protein. Although not tested, this antibody is likely functional in immunohistochemist
Western blot using Rocklands anti-RLC of Smooth and Non-muscle Myosin antibody to detect vascular myosin (rat aorta, lane 1) but not cardiac myosin (mouse heart, lane2). Each lane was loaded with 35 µg of lysate. Arrowheads indicate the detection of both mono-phosphorylated (upper) and unphosphorylated (lower) forms of the protein. After blocking with 5% Normal goat serum and 0.5% BLOTTO in PBS, the membrane was probed with the primary antibody diluted in blocking buffer to 1:600 for 2 h at room temperature. The membrane was washed and reacted with a 1:10,000 dilution of IRDye800(TM) conjugated Gt-a-Rabbit IgG [H&L] MX (611-132-122) for 45 min at room temperature (800 nm channel, green). Molecular weight estimation was made by comparison to prestained MW markers in lane M (700 nm channel, red). IRDye(TM)800 fluorescence image was captured using the Odyssey Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.
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