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[western blotting] Abcam's ab9484 b-actin loading control vs casein blocking
Abcam strongly reccommend to block ab9484 b-actin loading control
antibody (ab9484) with BSA instead of milk. Does anyone know what milk
component is most likely to cause problems with this ab? I need to
block this antibody with casein, hence my question.
[western blotting] Abcam's ab9484 b-actin loading control vscasein blocking
On 8 Dec 2006 02:48:57 -0800, Lechu <[Only registered users see links. ]> wrote:
so far I know milk protein gives problems with phosphorylation ....
that is if you plan to detect phosphorylation of proteins, you may
have a large background with phospho specific antibodies .... in this
case, where you're using GAPDH, the data sheet says that there's a
decrease in signal .....
now, if I were to give you a pie in the sky hypothesis, which would be
a plausible explanation for this decrease in activity:
a. you're using a monoclonal ... ergo: single epitope
b. the milk casein kinase phosphorylates this epitope
c ergo, your ab will not bind to it very well.
,,, probably if you had a polyclonal, you could get away with milk blocking.
|ab9484 , abcam , bactin , blocking , blotting , casein , control , loading , western|
|Thread||Thread Starter||Forum||Replies||Last Post|
|Loading Control Calculations||KKM||Western Blot Forum||1||09-23-2009 01:09 PM|
|Methods Digest, Vol 19, Issue 7 to Lechu Casein blocking question||Jess||Protocols and Methods Forum||0||12-09-2006 04:51 AM|
|[western blotting] polyclonal vs monoclonal loading control||Lechu||Protein Forum||1||11-29-2006 08:44 PM|
|actin loading control?||Josh Mcelwee||C Elegans Forum||0||07-18-2005 03:30 PM|