Trypsin Proteomics Grade

• Trypsin Spin Columns, Proteomics Grade
• Trypsin, Proteomics Grade
• Trypsin Profile IGD Kit

Trypsin, Proteomics Grade Superior Sequence Coverage Without Ambiguous Peaks! Proteomics Grade Trypsin (T6567) shows higher proteolytic efficiency than competitors' products, generating more tryptic peptides leading to greater sequence coverage of your proteins of interest. Mass spectra are significantly simplified due to the reduced number of interfering autolytic peaks and their ambiguous adducts. (Concerns over the lack of internal calibrant peaks at low mass may be addressed by calibrating on matrix peaks that routinely appear in the mass spectrum under normal circumstances.)

Proteomics Grade Trypsin has been extensively purified from porcine pancreas to enable accurate and precise cleavage on the carboxylic acid side of Arg and Lys residues. The enzyme has been exhaustively processed by reductive methylation to minimize autolysis and chymotryptic activity quenched by TPCK (N-Tosyl-L-phenylalanine chloromethyl ketone) treatment. Further purification steps including affinity chromatography and lyophilization from dilute acid produce a highly-purified, high specific activity trypsin purposely suited for the demanding criteria of proteomics research, and is designed to function for either in-solution or in-gel digests. The enzyme is conveniently packaged in five 20 µg vials to ensure fresh enzyme is available for each use.

Features & Benefits:

• Highly purified. Eliminates contaminant peaks and non- tryptic fragments from spectra.
• Stabilized form retains high specific activity. Complete digest with less protease for economy and confidence in spectra.
• Application document included with in-gel digest example. Save time in methods training and development.
• Convenient packaging. Save time and reduce waste by using only amount needed.

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Trypsin Profile IGD Kit Fast, Efficient, Complete Digestion, from PAGE to MALDI. The Trypsin IGD Kit (PP0100) enables fast, efficient, and complete in-gel tryptic digest of up to 100 excised protein spots. Digested proteins are ready for MALDI-MS and require no additional preparation. Because the Trypsin IGD Kit contains Proteomics Grade Trypsin, a higher sequence coverage and fewer ambiguous autolytic peaks are observed in MALDI spectra.

The Trypsin IGD Kit contains Proteomics Grade Trypsin and is optimized for polyacrylamide gels stained with Coomassie, SYPRO® Orange or Ruby dyes. If silver staining is used, ProteoSilver™ Plus silver staining kit is recommended (Product Code PROT-SIL2).

Features & Benefits:

• Includes highly purified Proteomics Grade Trypsin. Trypsin displays only minimal autolytic activity and no chymotryptic activity, which simplifies resulting peptide analysis.
• Detailed protocol. Easy to follow protocol simplifies the process.
• Works with Coomassie, SYPRO Orange, SYPRO Ruby or ProteoSilver Stained gels. Allows user a choice of staining methods.
• Resulting samples are suitable for analysis by MALDI-MS. No further processing needed, saves time.

Components
  Proteomics Grade Trypsin
  Destaining Solution
  Trypsin Reaction Buffer
  Biotech Grade Acetonitrile
  Trypsin Solubilization Reagent
  Peptide Extraction Solution

From SDS-PAGE to MALDI . . .

A solution of 2.5 mg/mL E. coli in Cellular and Organelle Membrane Solubilizing Reagent of ProteoPrep™ Total Extraction Sample Kit (Product Code PROT-TOT) was sonicated, then reduced with tributyl phosphine and alkylated with iodoacetamide (Product Code PROTRA). 109 µg of protein was loaded on a 7 cm, pH 4-7 IPG strip (Product Code I2906), focused for 50,000 volt hours, then run on a 4-20% Tris-Glycine SDS PAGE gel at 150 volts for 70 min. The gel was then stained with EZBlue (G 1041). A random spot was then excised and tryptically digested using the ProteoProfile Typsin In-Gel Digest Kit (Product Code PP 0100).

The sample was desalted using a ZipTip™ C18 pipette tip from Millipore and eluted directly onto the MALDI target using the MALDI matrix (a-cyano-4-hydroxycinnamic acid, 10 mg/mL in 70% ACN, 0.03% TFA). MALDI analysis was performed in the reflection positive ion mode. The resulting monoisotopic masses were searched against the NCBI database at a tolerance of 150 ppm. The protein was identified as outer membrane protein 3a from E. coli with the matched peptides providing 30% sequence coverage.

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