Question regarding muscle tissue homogenization - Why rotate homogenates after homogenization? (Nov/17/2007 )
Hello everyone!
I've been looking for homogenization protocols for muscle tissue protein extraction for Western Blot.
I have found several protocols that seem to be used regularly and most protocols are very similar.
However I have noticed one major difference amongst some of the protocols and that is that in some protocols the homogenates are rotated at 4 degrees for 30-120 min after homogenization.
My question is why? I haven't been able to find an explanation for rotating the homogenates following homogenization.
Any thoughts on the matter?
/W
-Willemman-
QUOTE (Willemman @ Nov 17 2007, 07:32 AM)
Hello everyone!
I've been looking for homogenization protocols for muscle tissue protein extraction for Western Blot.
I have found several protocols that seem to be used regularly and most protocols are very similar.
However I have noticed one major difference amongst some of the protocols and that is that in some protocols the homogenates are rotated at 4 degrees for 30-120 min after homogenization.
My question is why? I haven't been able to find an explanation for rotating the homogenates following homogenization.
Any thoughts on the matter?
/W
I've been looking for homogenization protocols for muscle tissue protein extraction for Western Blot.
I have found several protocols that seem to be used regularly and most protocols are very similar.
However I have noticed one major difference amongst some of the protocols and that is that in some protocols the homogenates are rotated at 4 degrees for 30-120 min after homogenization.
My question is why? I haven't been able to find an explanation for rotating the homogenates following homogenization.
Any thoughts on the matter?
/W
you may offer the protocol you mean; I speculate that it is to extract myofibrills; total homogenate is rarely used and difficult to handle
-The Bearer-
QUOTE (The Bearer @ Nov 17 2007, 09:39 PM)
QUOTE (Willemman @ Nov 17 2007, 07:32 AM)
Hello everyone!
I've been looking for homogenization protocols for muscle tissue protein extraction for Western Blot.
I have found several protocols that seem to be used regularly and most protocols are very similar.
However I have noticed one major difference amongst some of the protocols and that is that in some protocols the homogenates are rotated at 4 degrees for 30-120 min after homogenization.
My question is why? I haven't been able to find an explanation for rotating the homogenates following homogenization.
Any thoughts on the matter?
/W
I've been looking for homogenization protocols for muscle tissue protein extraction for Western Blot.
I have found several protocols that seem to be used regularly and most protocols are very similar.
However I have noticed one major difference amongst some of the protocols and that is that in some protocols the homogenates are rotated at 4 degrees for 30-120 min after homogenization.
My question is why? I haven't been able to find an explanation for rotating the homogenates following homogenization.
Any thoughts on the matter?
/W
you may offer the protocol you mean; I speculate that it is to extract myofibrills; total homogenate is rarely used and difficult to handle
Hi,
The protocol consists of homogenization followed by rotation of the homogenate at 4 degrees for 60 min after which the homogenate is centrifuged and the supernatant is used for the western blot. Other protocols I have found skip the rotation step but are the same in all other steps.
/W
-Willemman-