Hi, I am totally new in this field. My lab usually uses 0.25M sucrose and 0.1mM EDTA to homogenize fresh livers from rat. I just wonder if this homogenate could be used for Western Blot. If not, how much tissue (weight) will be needed for it? Thanks!
0
2 replies to this topic
#2
mdfenko
-
- Active Members
-
- 2,251 posts
an elder
79
Excellent
Excellent
Posted 04 February 2009 - 01:28 PM
yes, but...
there will be a lot of proteins in a homogenate, your protein of interest needs to be in enough abundance that it can be picked out from the crowd (depends on sensitivity of antibody).
there will be a lot of proteins in a homogenate, your protein of interest needs to be in enough abundance that it can be picked out from the crowd (depends on sensitivity of antibody).
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
#3
yobou
-
- Active Members
-
- 86 posts
Enthusiast
0
Neutral
Neutral
Posted 13 March 2009 - 09:56 PM
I guess you should homogenize in ordinary lysis buffer with protease and phosphatase inhibitors on ice. It is better to homogenize large piece to be representative as some area may be fibrotic or cirrhotic .
