Sooner Scientific Catalog

P.O. Box 180 - Garvin, OK 74736 - 580-286-7047 Fax 580-286-4268

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• Thin Layer Peptide Mapping Electrophoresis (TLPME) System
  • Description
  • Applications and Results
  • Ordering/Price Information

Complete system includes:

• Highly efficient aluminum cooling platform with Teflon insulation
• Clamping system with inflatable nylon air bag
• Wick adapted buffer tanks
• Safety power leads
• Safety cover with electric interlock
• Coolant flow cut-off valve (power supply shuts down if coolant flow stops)
• Step-by-step laboratory protocol and installation instructions
• In-line air pressure regulator with gauge

Applications and Results - Applications of Peptide Mapping and Advantages of the TLPME System Over Other Techniques.

Peptide mapping is a powerful technique to help determine peptide structure and composition of proteins. Peptide maps or fingerprints of proteolysed proteins are usually obtained by resolution on either one-dimensional SDS-PAGE analysis (Cleveland), reverse-phase HPLC, or by two-dimensional separation on thin layer plates. Perhaps the most common applications of peptide mapping are: 1) to reveal identities between proteins suspected to be encoded by the same or related genes, 2) to prepare individual peptides to determine amino acid composition and sequence, and 3) to determine the precise location of amino acid residues that are post-transitionally modified by either fatty acid acylation, glycosylation, methylation, acetylation, or phosphorylation. Because the biochemist is often faced with the reality of obtaining only vanishingly small amounts of protein for analysis, it is often difficult or impossible to perform crucial experiments which reveal some of these important characteristics.

Two-dimensional separation of proteolytic digests using the TLPME system and chromatography on thin layer cellulose plates is a technique that is well-suited to solve at least some of these problems and has certain advantages over reverse-phase HPLC and SDS-PAGE. First, it is an extremely sensitive technique that requires only small amounts of metabolically labeled product (only a few dpm). Second, because digests are resolved in two dimensions, a variety of information is derived that often yields subtle but important clues about a given peptide that may help reveal its composition. Finally, because cellulose is an inert substance, the peptide material can be recovered for secondary analysis such as determining amino acid composition and sequence or determining the presence and position of phosphoamino acid residues.

The Sooner Scientific TLPME System features a clamping system and an inflatable nylon air bag (see figure 1) which removes excess liquid from the surface of the plate and prevents buffer from siphoning. This creates and maintains an ultrathin interface between the buffer and cellulose during high-voltage electrophoresis and allows uniform cooling because of close and even contact with the cooling surface.

RESULTS: (Figure 2) Two-Dimensional Peptide Maps of Methionine-Containing Tryptic Peptides of c-Jun\AP-1 and FAP p39. As a test of the identity between c-Jun\AP-1 and FAP p39, the proteins were subjected to tryptic peptide mapping. (A) A two-dimensional peptide map of c-Jun\AP-1. (B) FAP p39. (C) an equal mixture of the two proteins. Ascending arrowhead marks the origin of each plate. Electrophoresis was in the horizontal plane with the anode at the left. Small descending arrowheads mark the location of three FAP p39-specific peptides. Peptides A-G represent peptides common to both proteins. Approximately 24,000 cpm in each panel. Shown are fluorographs obtained after exposing each plate to preflashed film for 24 hours at -70C. (2)REFERENCES: (1) J.A. Cooper, B.M. Sefton, and T. Hunter. Detection and quantification of Phosphotyrosine in proteins. Methods Enzymol. 99:387-402 (1983)

(2) R. Chiu, W.J. Boule, J. Meek, T. Smeal, T. Hunter, and M. Karin. The c-Fos Protein Interacts with c-Jun\AP-1 to stimulate Transcription of AP-1 Responsive Genes. Cell, V.54, 541-552 (1988)

Ordering/Price Information - Please see page 10 or the index.

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