Isoelectric focusing under dissociating conditions for analysis of muscle protein from clinically normal dogs and Labrador retrievers with hereditary myopathy.
Protein profiles of whole homogenates of anconeus (slow twitch) and biceps femoris (fast twitch) muscles of clinically normal dogs and of Labrador Retrievers with hereditary myopathy (HM) were resolved on flat bed polyacrylamide isoelectric-focusing gels. Three methods of sample solubilization were performed. The solubilization buffer, with high concentrations of urea, precipitated the zwitterionic detergent, but use of the buffer containing 3% NP-40, 9.2M urea, and 0.1M arginine resulted in better resolution and stability of pH gradient. Gels of anconeus muscle from clinically normal dogs contained 2 protein bands specific to anconeus muscle, whereas gels of biceps femoris muscle from clinically normal dogs contained 3 protein bands amplified in biceps femoris muscle that were barely detectable in anconeus muscle. The staining intensity of protein bands in biceps femoris muscles from Labrador Retrievers with HM was decreased, relative to controls. The quantitative analysis of peak height ratios of biceps femoris muscle revealed significant (P less than 0.05) differences between profiles of clinically normal dogs and Labrador Retrievers with HM.
Mehta
J R
JR
Neuromuscular Laboratory, College of Veterinary Medicine, Auburn University, AL 36849.
Braund
K G
KG
McKerrell
R E
RE
Toivio-Kinnucan
M
M
D000818:Animals; D003720:Densitometry; D004283:Dog Diseases / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D004285:Dogs; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D009124:Muscle Proteins / Q000032:analysis*; D009132:Muscles / Q000032:analysis / Q000473:pathology; D009135:Muscular Diseases / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000662:veterinary*
D009124:Muscle Proteins