Lyophilization, a practical way to store and transport tissues prior to protein extraction for 2DE analysis?
To date, lyophilized samples are rarely utilized in proteomics experiments. This is most likely because researchers are concerned about inducing cross-linking of proteins via amide bonds, leading to artefactual charge modification and thus resulting in irreproducible results and bad gels. Indeed, it is known that lyophilization can cause crosslinking. The potential reaction is a reaction of free amino groups of a protein (N-terminal alpha-amino groups and epsilon-amino groups from lysine) with the reducing group of sugar molecules. The rate and extent of this reaction depends on the sugar content of the sample, the efficiency of lyophilization process, the residual water content of the material and the storage temperature. Lyophilization is a cheap, practical and safe alternative for the storage and transportation of samples prior to protein extraction, separation and quantification via 2DE, when care is taken (i) to dry the samples to the lowest practicable moisture content, (ii) to transport and store them under water- and airtight conditions and (iii) to avoid heating of the sample.
Carpentier
Sebastien C
SC
Faculty of Bioscience Engineering, K. U. Leuven, Leuven, Belgium. sebastien.carpentier@biw.kuleuven.be
Dens
Koen
K
Van den houwe
Ines
I
Swennen
Rony
R
Panis
Bart
B
D002241:Carbohydrates / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005612:Freeze Drying / Q000379:methods*; D008239:Lysine / Q000737:chemistry; D028521:Musa / Q000737:chemistry; D018515:Plant Leaves / Q000737:chemistry; D010940:Plant Proteins / Q000302:isolation & purification; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D013048:Specimen Handling / Q000379:methods
D002241:Carbohydrates; D010940:Plant Proteins; D011506:Proteins; D008239:Lysine