changes in protein profiles in bortezomib applied multiple myeloma ...

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