Efficient micro-recovery and guanidination of peptides directly from MALDI target spots.
A method is presented for the recovery and subsequent guanidination of tryptic peptides from samples previously spotted on a matrix-assisted laser desorption/ionization (MALDI) target. The procedure is shown to have applicability to both in-solution and in-gel digests, yielding improved confidence in protein identification and sequence coverage in all instances. Recovery from the plate is essentially quantitative, with no residual analyte observed on the target spot. The technique is rapid, simple, and has extended applicability to other processing steps, including (but not limited to) derivatization for specific peptide studies or enzymatic treatment for subsequent profiling of posttranslational modifications. This method circumvents the failure of an initial analysis to generate suitable information and is particularly relevant for the analysis of precious samples.
Cockrill
Steven L
SL
Sigma-Aldrich Corporation, St. Louis, MO, USA.
Foster
Kelly L
KL
Wildsmith
Justin
J
Goodrich
April R
AR
Dapron
John G
JG
Hassell
Tom C
TC
Kappel
William K
WK
Scott
Graham B I
GB
D019791:Guanidine / Q000737:chemistry*; D008832:Microchemistry / Q000379:methods*; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification*; D020539:Sequence Analysis, Protein / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D010455:Peptides; D019791:Guanidine