Development of mini-gel technology in two-dimensional electrophoresis for mass-screening of samples: application to tears.
Despite the extensive literature available on tear proteins and lipids, very little has been reported on the tear fluid as a whole and it's changes in contact lens wear or ocular diseased patients. Initially a human reflex tear two-dimensional map was created by Molloy et al. (Electrophoresis 1997, 18, 2811-2815), using this information a process for mass-screening was established. The large format two-dimensional technique was evaluated, using a basal tear reference map, and modified to describe a fast, efficient and cost effective method of protein separation. The use of one pH 3-10 18 cm nonlinear immobilised pH gradient (IPG) strip and two mini-gels for the second-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) results in an effective separation of tear proteins which will be applied in diagnostic studies of tear samples.
Glasson
M J
MJ
Co-operative Research Centre for Eye Research and Technology, UNSW, Sydney, Australia.
Molloy
M P
MP
Walsh
B J
BJ
Willcox
M D
MD
Morris
C A
CA
Williams
K L
KL
D000180:Acrylic Resins; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D011506:Proteins / Q000302:isolation & purification*; D013666:Tears / Q000737:chemistry*
D000180:Acrylic Resins; D011506:Proteins; C016680:polyacrylamide gels