2D gel or MS? for comparison proteomic

shifali chatrath via methods%40net.bio.net (by shifalich from rediffmail.com)
Thu Mar 5 00:22:48 EST 2009


  
Yeah! I later checked with my colleagues.He also told me the same thing DIGE. But he also told its very cumbersome!

Shifali

On Thu, 05 Mar 2009 Dr Engelbert Buxbaum wrote :
>Am 26.02.2009, 08:56 Uhr, schrieb shifali  chatrath  <shifalich from rediffmail.com>:
>
>
>>we do all kinds of proteomic techniques. Also, a lot of mol. Bio. stuff.  and as far as I know, 2D and MS/MS can only be used you define the  composition of proteome in two types of cells (If you meant so....) but  not to quantitate.
>
>Oh yes, 2D-electrophoresis can be used to compare levels of proteins in  different samples, this is called "differential gel electrophoresis"  (DIGE). From each sample you take an aliquote and label it with  fluorescent dyes (or stable isotopes). An additional dye is used to label  a mixture of all samples (internal standard). All samples are then mixed  and run on the same gel. Then the fluorescence of a spot at different  wavelengths is measured, the ratio is proportional to the ratio of  proteins in the samples.
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Shifali Chatrath
Graduate Student
Protein science Lab
Dept. of Biological sciences
National University of Singapore
Singapore
+65-65161210
+65-96393449


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