changes in protein profiles in bortezomib applied multiple myeloma ...
changes in protein profiles in bortezomib applied multiple myeloma ...
changes in protein profiles in bortezomib applied multiple myeloma ...
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Urea (8M) 4.8 g<br />
CHAPS (%2) 0.2 g<br />
DTT (50 mM) 0.077 g<br />
Ampholites (3-10) (%0.2) 100 µl<br />
Bromophenol Blue (%0.5) 1-2 µl<br />
Preparation of Rehydration Buffer<br />
Total<br />
Volume<br />
= 10 ml<br />
For urea was <strong>in</strong>creas<strong>in</strong>g the volume of liquid when dissolved, firstly, it<br />
was dissolved <strong>in</strong> 4ml ddH2O. Then the other chemicals were added, total<br />
solution was measured to see its volume and the f<strong>in</strong>al volume was<br />
br<strong>in</strong>ged to 10 ml.<br />
Ampholites was diluted to 1X from the 10X stock concentration. The<br />
ampholites concentration <strong>in</strong> total volume is %0.2.<br />
Nearly, 1-2 µl Bromophenol Blue (%0.5) was used just to have color <strong>in</strong><br />
buffer.<br />
B.6.1.2. Preparation of Equilibration Buffer I and II Stock<br />
Equilibration Buffer I (20 ml) Equilibration Buffer II (20 ml)<br />
7.2 g / 20 ml Urea 7.2 g / 20 ml Urea<br />
0.4 g / 20 ml SDS 0.4 g / 20 ml SDS<br />
5 ml Ultrapura H2O 5 ml Ultrapura H2O<br />
5 ml 1.5 M Tris – HCl pH = 8.8 5 ml 1.5 M Tris – HCl pH = 8.8<br />
0.4 g / 20 ml DTT -<br />
4 ml Glycerol 4 ml Glycerol<br />
- 0.5 g / 20 ml Iodoacetamide*<br />
* Iodoacetamide should be added freshly. (<strong>in</strong> Day 3).<br />
After preparation, Equilibration Buffer I was lifted to +4 o C while Equilibration<br />
Buffer II was kept at -20 o C.<br />
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