Proteomics

[Diwakar]

Hi

I have problems in preparing a xylem sap sample from tomato for analyzing the total proteome. Does any of you can advise me?

[elle]

what is your problem?

[danfive]

Try reading the article cited below:

Originally Posted by danfive

I just wanted to add this great article, "Reduction of proteins during sample preparation and two-dimensional gel electrophoresis of woody plant samples," as a reference for people interested in plant protein extraction and 2D gels, etc.

The link --> Wiley InterScience :: Session Cookies

[Diwakar]

Actually there are many small problems. The main problem is that i am not getting enough concentration of plant proteins (which should be) from infected tomato plants (with Ralstonia solanacearum).I have concentrated 30 mL-50 mL xylem sap to obtain 100-200 µg of protein but many times failed. I even tried lyophilizer to concentrated sap but then the powder obtained couldn,t be dissolved in sample buffer to run for gel electrophoresis. IF anybody has sugestion then please forward to me.

Thanks

[danfive]

Check out the methods in this article: http://www.springerlink.com/content/...3/fulltext.pdf

Maybe their acetone:methanol: DTT precipitation method will work for you.