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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B.5. APO LOGIX JC-1 Mitochondrial Membrane Potential Detection<br />
Kit<br />
APO LOGIX JC-1 Mitochondrial Membrane Potential Detection Kit was<br />
purchased from Cell Technology, USA.<br />
B.5.1. JC-1 Reagent<br />
The lyophilized vial was reconstituted with 500 μl DMSO to obta<strong>in</strong> a 100X<br />
stock solution. Follow<strong>in</strong>g that, it was mixed by <strong>in</strong>vert<strong>in</strong>g several times at room<br />
temperature until contents were completely dissolved. Next, resuspended JC-1 reagent<br />
was aliquated <strong>in</strong> small amountssufficient for one day experimental work and stored at -<br />
20 o C. When it was used, it was diluated 1:100 <strong>in</strong> 1X Assay Buffer (100X JC-1 reagent<br />
to 1X) immediately prior to use.<br />
B.5.2. 10X Assay Buffer<br />
10X Assay Buffer were diluted 1:10 with ultrapure water (1 ml 10X Assay<br />
Buffer + 9 ml UP Water).<br />
B.6. 2D PAGE<br />
B.6.1. Isoelectric Focus<strong>in</strong>g (IEF)<br />
B.6.1.1. Preparation of Rehydration Buffer<br />
It was used for rehydration of strips.<br />
This protocol was for <strong>in</strong>clud<strong>in</strong>g prote<strong>in</strong>s <strong>in</strong> buffer.<br />
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