Canine Pancreatic Microsomal Membranes Technical Manual

Literature # TM231

Microsomal vesicles are used to study cotranslational and initial post-translational processing of proteins. Processing events such as signal peptide cleavage, membrane insertion, translocation and core glycosylation can be examined by the translation of the appropriate mRNA in vitro in the presence of these microsomal membranes. In addition, processing and glycosylation events may be studied by transcription/translation of the appropriate DNA in the TNT® Lysate Systems when used with microsomal membranes. To assure consistent performance with minimal translational inhibition and background, microsomes have been isolated free from contaminating membrane fractions and stripped of endogenous membrane-bound ribosomes and mRNA. Membrane preparations are assayed for both signal peptidase and core glycosylation activities using two different control mRNAs. The two control mRNAs supplied with this system are the precursor for beta-lactamase (or ampicillin resistance gene product) from E. coli and the precursor for alpha-mating factor (or alpha-factor gene product) from S. cerevisiae.