Important Sample Preparation Considerations



Sample handling guidelines for various sample types


Gel spots or bands: Excised gel bands can easily be contaminated with other proteins from direct contact with your hands when preparing the sample. To minimize contamination, please use nitrile gloves when handling the gel bands and keep ALL EQUIPMENT, which contacts the gel band clean by rinsing with HPLC grade water (including the eppendorf tubes used for the samples). Ship in DRY ICE.

Liquid samples: Wrap sample vial caps (flip top or screw cap) with ParaFilm to avoid inadvertent loss of material from the sample container. Ship in DRY ICE.

Frozen samples: Snap freeze samples using liquid nitrogen for efficient sample preservation. Please do not fill sample vials over 80% to avoid fracturing of the plastic vial by expansion of the sample material. When freezing the samples, please be sure they are in an upright position to keep the sample material away from the lid/cap. Ship in DRY ICE.

Cultured cells (suspension): Rinse cells, sequentially in 2-3 volumes of PBS to remove any media related protein or other soluble materials from the cultured cells. Pellet in an isotonic buffer such as PBS to prevent lysis. Flash freeze the cell pellet after rinsing the cells. Ship in DRY ICE.

Cultured cells (adherent): Rinse cells, sequentially in 2-3 volumes of PBS to remove any media related protein or other soluble materials from the cultured cells. Scrape cells in UREA LYSIS BUFFER (8 M urea, 20 mM HEPES pH 8.0, 1 mM beta-glycerophosphate, 1 mM sodium orthovanadate (activated), 2.5 mM sodium pyrophosphate, 1 mM DTT, 1 mM EDTA) and transfer to 50 mL Falcon tube. Flash freeze the cell slurry in liquid nitrogen. Ship in DRY ICE.

NOTE: Prepare all buffers and stock solutions with Milli-Q (or equivalent grade) purified waters. Do not use any labware that has been exposed with detergents since these materials can interfere with the LC and MS performance.

Tissue samples: Snap freeze tissue samples directly in liquid nitrogen for efficient sample preservation. Ship in DRY ICE.

© 2015 Lighthouse Proteomics LLC. All Rights Reserved.
Designed by Peak Design Firm