Guidelines

Preparation of protein samples separated by SDS-PAGE and delivered in gel form or blotted onto PVDF membrane:

• Work always with gloves, on dust-free surfaces, and with clean tools in order to avoid any contamination with keratin. Rinse/wash all surfaces and tools with 70% ethanol.
• De-staining of Coomassie stained gels to a clear background can be an advantage. For silver-staining, stain only as long until bands of interest appear (usually only a few minutes); avoid over-staining!
• If possible, take a picture of the gel prior to excision of gel bands and submit an image along with the sample.
• Cut gel bands with scalpel accurately along stained region. Please cut only a vertical segment in the middle of the band in case of a fat and broad gel band.
• Please avoid any gel pieces from the stacking/separation gel boundary. Please note that low % acrylamide gel pieces (stacking gel) do block our LC setup!
• Slice bands into small (~1 mm3) cubes. Do not deliver more than 6 small cubes.
• Transfer gel cubes into a clean, keratin-free 1.5 ml-reaction vial with lid (Eppendorf type). Please NO tubes with screw caps!
• Cover gel cubes with liquid composed of ethanol/water 20:80.  Pieces of membranes are kept dry. Under this condition, samples can be stored for several weeks at -20° or 4°C.
• Label tubes with permanent ink with your name and the sample identifier. Please do not label the lid!
• Keep samples at 4°C and bring them to our laboratory Murtenstrasse 28, level 4, Laboratory 474

Thank you!