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 The CNRU Mass Spectrometry Core Facility is equipped with today's most advanced scientific equipment.
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RECIPES FOR SAMPLE BUFFERSThe following are recipes for the buffers typically used to load samples onto the isoelectric focusing strips. These buffers may also be used to lyse cells. At the CNRU Mass Spectrometry Core Facility we typically first try the Chaotropic Buffer for our sample preparation.CHAOTROPIC BUFFER
2M Thiourea (760 mg) *50 mM DTT 4% CHAPS (100 mg) **0.2% ampholytes Nanopure H2O to 5ml STANDARD BUFFER
*50 mM DTT (385 mg) 4% CHAPS (2g) **0.2 % Carrier ampholytes Nanopure (adjust to 50 ml) MODIFIED O'FARRELL BUFFER(Gorg et al 2000)9M Urea (27g dissolved in 25 ml water) *1% DTT (0.5 g) 4% CHAPS (2g) **0.2% Ampholytes Nanopure to 50 ml AMPHOLYTESUse pH 3-10 for all ranges except pH 6-11 (in which case use Amersham's 6-11 ampholytes). Ampholytes should have a final concentration of 0.2%. For 1 mL of buffer, add 5 ul of ampholytes. For 5 mL of buffer, add 25 ul of ampholytes.* DTT is added just prior to use ** Carrier ampholytes will be added by us just prior to focusing. |
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