Bio-Rad ReadyPrep™ 2-D Starter Kit - Manual (Page 3)

Section 1
Introduction
The ReadyPrep ™ 2-D starter kit was designed as a single-use kit to familiarize first-time users with
the utilization of the Bio-Rad PROTEAN ® IEF cell and ReadyStrip ™ IPG strips. The kit provides all
the reagents necessary, including a protein control, to successfully perform two-dimensional poly-
acrylamide gel electrophoresis (2-D PAGE). The kit contains sufficient reagents to perform 2-D
PAGE of at least 6 samples with any of the three SDS-PAGE formats Bio-Rad offers. The amount
of sample applied to each gel is sufficient so that a large number of protein spots can be readily
detected when using either Bio-Safe
™ Coomassie* stain or Coomassie Brilliant Blue R-250 stain.
This manual provides instructions on how to perform 2-D PAGE with ReadyStrip IPG strips using
the Bio-Rad PROTEAN IEF cell and the control E. coli protein sample.
Section 2
Kit Components
ReadyPrep E. coli Protein Sample. One vial. Lyophilized. Each vial of reconstituted
ReadyPrep E. coli protein sample contains 2.7 mg of total protein at a concentration of 1.35
mg/ml.
ReadyPrep Rehydration/Sample Buffer. One vial. Lyophilized. Each vial of reconstituted
ReadyPrep rehydration buffer contains 10 ml of 8 M urea, 2% CHAPS, 50 mM dithiothreitol (DTT),
0.2% (w/v) Bio-Lyte
® 3/10 ampholytes, and Bromophenol Blue (trace).
Nanopure Water. One bottle containing 15 ml of sterile nanopure water.
Equilibration Buffer I. Two vials, each containing a stirbar. Lyophilized. Each vial of reconstitut-
ed equilibration buffer I contains 20 ml of 6 M urea, 2% SDS, 0.375 M Tris-HCl (pH 8.8), 20%
glycerol, and 2% (w/v) DTT.
Equilibration Buffer II. Two vials, each containing a stirbar. Lyophilized. Each vial of reconsti-
tuted equilibration buffer II contains 20 ml of 6 M urea, 2% SDS, 0.375 M Tris-HCl (pH 8.8), and
20% glycerol.
30% Glycerol Solution. One bottle containing 70 ml of sterile 30% (v/v) glycerol.
Iodoacetamide. Two vials. Each vial contains 0.5 gr of an ultrapure grade of iodoacetamide.
*Coomassie is a trademark of Imperial Chemical Industries PLC.
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