A method for application of samples to matrix-assisted laser desorption ionization time-of-flight targets that enhances peptide detection.

Item request has been placed! ×
Item request cannot be made. ×
loading   Processing Request
Read More Add to Saved list
  • Author(s): Landry F;Landry F; Lombardo CR; Smith JW
  • Source:
    Analytical biochemistry [Anal Biochem] 2000 Mar 01; Vol. 279 (1), pp. 1-8.
  • Publication Type:
    Journal Article; Research Support, U.S. Gov't, P.H.S.
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Elsevier Country of Publication: United States NLM ID: 0370535 Publication Model: Print Cited Medium: Print ISSN: 0003-2697 (Print) Linking ISSN: 00032697 NLM ISO Abbreviation: Anal Biochem Subsets: MEDLINE
    • Publication Information:
      Publication: <2000- > : San Diego, CA : Elsevier
      Original Publication: Orlando Fl : Academic Press
    • Subject Terms:
      Peptides/*analysis ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/*methods; Animals ; Cattle ; Detergents ; Humans ; Membrane Proteins/analysis ; Membrane Proteins/isolation & purification ; Peptide Fragments/analysis ; Peptide Fragments/isolation & purification ; Peptides/isolation & purification
    • Abstract:
      Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry has become a fundamental tool for the identification and analysis of peptides and proteins. MALDI-TOF is well suited for the analysis of complex biological mixtures because samples are crystallized onto a solid support that can be washed to remove contaminants and salts prior to laser desorption. A number of approaches for immobilizing samples onto MALDI targets have been put forth. These include the use of different chemical matrices and the immobilization of samples onto different solid supports. In large part though, the preparation of MALDI targets has been an empirical exercise that often requires a unique series of conditions for every sample. Here, a simple method for the application of peptide mixtures onto MALDI targets is put forth. This method differs because peptides are added directly to a sample of nitrocellulose dissolved in acetone, allowing them to interact in solution-phase organic solvent. This solution-phase mixture is then spotted to the MALDI target and evaporated, forming a homogenous solid surface for laser desorption. This procedure is robust, highly sensitive, tolerant to detergents, and easily learned. In our hands, the method provides as much as a 10-fold enhancement to the detection of tryptic peptide fragments derived from in-gel digests.
      (Copyright 2000 Academic Press.)
    • Grant Information:
      AR42750 United States AR NIAMS NIH HHS; CA30199 United States CA NCI NIH HHS; CA69306 United States CA NCI NIH HHS; etc.
    • Accession Number:
      0 (Detergents)
      0 (Membrane Proteins)
      0 (Peptide Fragments)
      0 (Peptides)
    • Publication Date:
      Date Created: 20000223 Date Completed: 20000508 Latest Revision: 20071114
    • Publication Date:
      20221213
    • Accession Number:
      10.1006/abio.1999.4468
    • Accession Number:
      10683224