help me to prepare urine protein sample preparation in 2D gel Electrophoresis

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nalpy
nalpy's picture
help me to prepare urine protein sample preparation in 2D gel Electrophoresis

Can any one send the details protocol to prepare urine protein sample preparation in 2D gel Electrophrosis and already not not work in crude protein sample.

pundir
pundir's picture
Hi nalpy,


Hi nalpy,
you can find some relevent information in the articles mentioned below
"Systematic Evaluation of Sample Preparation Methods for Gel-Based Human Urinary Proteomics: Quantity, Quality, and Variability" pubs.acs.org/doi/pdf/10.1021/pr0502525

 
"Proteomic analysis of normal human urinary proteins isolated by acetone precipitation or ultracentrifugation"  www.nature.com/ki/journal/v62/n4/pdf/4493245a.pdf

ARGERINE
ARGERINE's picture
Hi Nalpy

Hi Nalpy
There are alot of published research articles and reviews in the field of Urinary proteome analysis. Especially During the HUPOs plasma proteome project in Proteomics and Journal of proteomics as well. I have alot of those articles with me as Im working in the field of plasma proteomics.
Few of the key links are
http://jasn.asnjournals.org/cgi/reprint/18/4/1057
http://www.mcponline.org/cgi/content/full/5/10/1760
http://ajprenal.physiology.org/cgi/reprint/287/3/F353.pdf
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2215359
http://www3.interscience.wiley.com/journal/107629553/abstract?CRETRY=1&SRETRY=0
In case you dont have access to these journals send me your email ID I will enclose the articles to you.
All the best

nalpy
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Dear Pundir & Argerine

Dear Pundir & Argerine
Thank you for your kind suggestion and papers link

varsha
varsha's picture
Hi Nalpy.

Hi Nalpy.
I knew a student who did it. It involves acetone pptn of the urine sample. Then you can up the ppt in PBS  for protein estimation or directly into 2D lysis buffer. The protocol is described in the following paper if you can get it from the library . Good luck!
Varsha

nalpy
nalpy's picture
Dear  Dr. Varsha

Dear  Dr. Varsha
Thank you for your kind help and follow the protocol.
Regards
palanisamy