Cell Signaling Canine Pancreatic Microsomal Membranes Product Canine Pancreatic Microsomal Membranes For Research Use Only. Not for Use in Diagnostic Procedures. Description: Microsomal vesicles are used to study co-translational 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 the transcription/translation of the appropriate DNA in the TNT® Lysate Systems when used with Canine Pancreatic 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 β-lactamase (or ampicillin resistance gene product) from E. coli and the precursor for α-mating factor (or α-factor gene product) from S. cerevisiae. The Signal Sequence Control mRNA (E. coli β-lactamase) is transcribed by SP6 RNA polymerase from a plasmid bearing the coding region for the E. coli gene encoding the precursor to β-lactamase (the ampicillin resistance gene product). The RNA is synthesized without a cap analog. This control mRNA is used to assay for signal peptidase activity and is supplied with the Canine Pancreatic Microsomal Membranes System. The Core Glycosylation Control mRNA (S. cerevisiae α-factor) is transcribed by SP6 RNA polymerase from a plasmid bearing the coding region for the S. cerevisiae α-mating factor. The RNA is synthesized without a cap analog. This control mRNA is used to assay for core glycosylation activity and is supplied with the Canine Pancreatic Microsomal Membranes System. Features: • Reliable: Microsomes are stripped of endogenous membrane-bound ribosomes and mRNA to ensure consistent performance with minimal translational inhibition and background. Performance tested in rabbit reticulocyte lysate. Storage Conditions: Store at –70°C or below. Membranes are stable at –70°C for 1 year. After thawing, unused portions should be rapidly refrozen in liquid nitrogen. No detectable loss of activity results after two freeze-thaw cycles. Protocol Canine Pancreatic Microsomal Membranes Technical Manual Part# TM231 Size Cat.# Price (Fr) 50 µl Y4041 588.00 µl CMM precursor (~31.5kDa) processed (~28.9kDa) Signal Processing 0 0.3 0.6 0.9 1.2 1.5 1.8 2.1 2.4 13 µl CMM processed (~30.0kDa) 0 0.3 0.6 0.9 1.2 1.5 1.8 2.1 2.4 Glycosylation precursor (~18.6kDa) Processing and glycosylation activity of Canine Pancreatic Microsomal Membranes (CMM). The positive control mRNAs (0.5µg each of E. coli β-lactamase and S. cerevisiae α-factor) were translated using Rabbit Reticulocyte Lysate in a 25μl reaction for 60 minutes in the presence of the indicated amounts of CMM (3μl). Translation products were analyzed by gel electrophoresis followed by autoradiography of the [35S]-labeled proteins. Amino Acid Mixtures Product Amino Acid Mixture, Complete Amino Acid Mixture Minus Cysteine Amino Acid Mixture Minus Methionine and Cysteine Amino Acid Mixture Minus Leucine Amino Acid Mixture Minus Methionine For Laboratory Use. Size Conc. Cat.# Price (Fr) 175 µl 1 mM L4461 93.00 175 µl 1 mM L4471 93.00 175 µl 1 mM L5511 93.00 175 µl 1 mM L9951 93.00 175 µl 1 mM L9961 93.00 Description: The Amino Acid Mixture, Complete, is an aqueous solution containing 1mM each of the 20 essential amino acids. This mixture is compatible for use in the Flexi® Lysate, TNT® Lysate and standard Rabbit Reticulocyte Lysate Systems as well as in the Wheat Germ Extract and E. coli S30 Systems. Amino Acid Mixtures are also available lacking cysteine, methionine and cysteine, leucine or methionine. Storage Conditions: Store at –70°C. For complete and up-to-date product information visit: www.promega.com/catalog 229 Protein Expression and Analysis 0373TA07_3B Pagina 232
Pagina 234Heeft u een onderzoeksrapport, onlinemag of online flyers? Gebruik Online Touch: artikel naar een online publicatie omzetten.
Promega Switzerland - Life Sciences Catalog 2012 Lees publicatie 100Home