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iTRAQ cost compared to a DIGE approach - (Nov/03/2009 )

Dear all,

Iím thinking about doing a non-gel based technique approach to identify cell wall proteins (most of them are basic and highly glycosilated). Because of these characteristics, proteins doesnít go easily inside of isoelectric focusing gels, so we will lose a lot of such proteins in a 2D approach.

Iím thinking about using iTRAQ technology, but until this point I donít know how high would be the cost for analyzing 3 control and 3 treated samples. Do someone of you know, if the cost of such non-gel based techniques are much higher than a usual DIGE experiment? I refer to the cost of chemicals.

Thank you for your support.

-kern-

Dear Kern,

I am also planning to use iTRAQ in my lab. What I have understood after going through different protocols, quotations etc is that iTRAQ is very expensive!

Also let me ask you one questions: Are you planning to outsource the samples or do you have all the necessary equipments in your lab? I am asking oyu this because this factor has got a huge impact on the cost of iTRAQ.

1. iTRAQ kits can cost you from $1200 to around $2000 (depends what kind of kit you want - 4-plex or 8-plex). This kit is the basic necessity that you can not avoid. After iTRAQ labelling comes the processing of these labelled-proteins/peptides and then comes analysis.

2. I think Strong Cation-Exchange Chromatography is the best method to separate when using iTRAQ, though SDS-PAGE and subsequent in-gel digestion etc. is also found to be used.

3. LC-MS/MS analysis after SCX fractionation can also be costly depending upon the number of samples/fractions.

By the way, what protocol are you going to use? Have you come across any interesting protocol that is neatly written, then I would request you to share it so that everybody can benefit from it.

I am facing many technical challenges regarding iTRAQ-SCX. So, I will be posting some questions on that here. Please contribute to that in all the ways possible.

Thanks

K.

-Neanderthal-