UCDHSC Anesthesiology Research - Clinical Nutrition Research Unit

The CNRU Mass Spectrometry Core offers Proteomics and Metabolomics workshops.

RECIPES FOR SECOND DIMENSION

AGAROSE SEALANT

1% agarose in running buffer. Boil gently until agarose is in solution, then add 0.002% bromophenol blue and aliquot. Store at 4°C.

BUFFERS

Equilibration Stock Solution

	10 mL	100 mL	500 mL
Urea (6M)	6 g	36 g	180g
1.5 M Tris pH 8.8 (0.05M)	0.33 ml	3.3 mL	16.5 mL
20% SDS (2%)	1 mL	10mL	50 mL
Glycerol (20%)	2 mL	20 mL	100 mL
Water		Adjust to 100mL	Adjust to 500 ml

Add DTT (100mg per 10mL) and IAA (250 mg per 10mL) fresh just prior to using. SDS Running Buffer
10L Tris Running Buffer (25mM tris, 192mM glycine, 0.1% SDS) 30.28g Tris
144.1g glycine
50ml of 20% SDS

20L Tris Running Buffer (25mM tris, 192mM glycine, 0.1% SDS)
60.56g Tris
288.2g Glycine
100ml of 20%SDS

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