Go Back   Molecular Biology Forum > Molecular Research Topics Forum > Proteomics Forum
Register Blogs FAQ Members List Calendar Science Groups New! Arcade Search Today's Posts Mark Forums Read

Proteomics Forum Post Questions and Discuss Proteomics, Proteomic Bioinformatics, Proteomic Techniques such as 2-D, Mass Spec etc.


Protein extraction from rabbit hearts

Proteomics Forum

Post Questions and Discuss Proteomics, Proteomic Bioinformatics, Proteomic Techniques such as 2-D, Mass Spec etc.



Register Molecular Biology Forums
Reply
 
LinkBack Thread Tools Display Modes
  #1 (permalink)  
Old 10-25-2006, 07:17 PM
Pipette Filler
Points: 1,089, Level: 18Points: 1,089, Level: 18Points: 1,089, Level: 18
Activity: 0%Activity: 0%Activity: 0%
 

Join Date: Oct 2006
Posts: 3
Amiodaron RSS Feed
Question Protein extraction from rabbit hearts

Hi!
We are planning to apply proteomics to unveil cardiomyocyte protein expression patterns in distinct stages of heart failure. We want to use a rabbit model of heart failure. So does anyone know any protocols regarding the first steps of the proteomic-workflow in this specific experimental setting: retrograd perfusion of the heart, cardiomyocyte isolation, protein extraction...?

Thanks
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
Alt Today
Advertising
Google Adsense
 
This advertising will not be shown
in this way to registered members.
Register your free account today
and become a member on
Molecular Biology Forum
Standard Sponsored Links

  #2 (permalink)  
Old 10-26-2006, 12:50 AM
kiki06's Avatar
Post-Doc
Points: 4,288, Level: 44Points: 4,288, Level: 44Points: 4,288, Level: 44
Activity: 0%Activity: 0%Activity: 0%
 
Join Date: Feb 2006
Posts: 263
kiki06 RSS Feed
Lightbulb Re: Protein extraction from rabbit hearts

Wow! welcome to the forum. That is a pretty complicated procedure although certainly doable.

1) You would have to isolated the cardiomyocytes carefully.
This would entail looking up Pubmed to see what previous investigators did to extract viable cardiomyocytes.
Q: Do you really need cardiomyocytes carefully extracted? Or can you simply remove the heart and simply extract the proteins from whole tissue?


2) Protein extraction is quite easy once you got the cells. You could easily extact with a proteomics compatible lysis buffer. Dialyze the cells and get them ready in a buffer that allows 2-D separation. Hard part is going to be getting SAME amount of protein loaded for DISEASE heart failure cells and Normal cells.

3) Separate on 2-D. Get a fun robot to collect the spots. Send them all in for sequencing! and compare using scanning/sequencing CONTROL vs DISEASE!

hope that helps!
post here your response!

Kiki
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
  #3 (permalink)  
Old 10-26-2006, 07:27 AM
Pipette Filler
Points: 1,089, Level: 18Points: 1,089, Level: 18Points: 1,089, Level: 18
Activity: 0%Activity: 0%Activity: 0%
 

Join Date: Oct 2006
Posts: 3
Amiodaron RSS Feed
Default Re: Protein extraction from rabbit hearts

Hi Kiki,

thank you for your response. I have already scanned the available literature (there are just a few research groups dealing with that specific experimental setting), but the procedural approach here is quite different. It is indeed an important question, if the isolation of cardiomycytes is really necessary or if it would not be easier to prefer a whole tissue-extract. The problem of whole tissue-extracts thereby is the unavoidable contamination by other cardiac cells (fibroblasts, leukocytes, endothelial cells, vascular smooth muscle cells...), so altered protein expression patterns in cardiac tissue-extracts do not necessarily reflect an altered cardiomyocyte protein-expression status. Nevertheless, some scientist have chosen that way...

best regards
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
Reply

Tags
extraction , hearts , protein , rabbit

Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On
Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Plant Protein Extraction the Hard Way danfive Protein Science 0 04-24-2008 08:53 PM
Protein Extraction Problems molecule2005 Protein Science 0 04-16-2008 08:57 AM
Production of Monoclonal Antibodies - Technique in Mouse Cellbiogal Antibody Forum 1 02-17-2008 02:00 PM
Nuclear Protein Extraction Mossaded Protocols and Methods Forum 1 06-12-2007 08:56 PM
Nuclear Protein Extraction Mossaded Protein Science 0 06-12-2007 02:27 PM


All times are GMT. The time now is 12:04 AM.


Powered by vBulletin® Version 3.7.1
Copyright ©2000 - 2008, Jelsoft Enterprises Ltd.
Copyright 2005-2007 Molecular Station | All Rights Reserved