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[western blotting] Abcam's ab9484 b-actin loading control vs casein blocking

[western blotting] Abcam's ab9484 b-actin loading control vs casein blocking - Protocols and Methods Forum

[western blotting] Abcam's ab9484 b-actin loading control vs casein blocking - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 12-08-2006, 10:48 AM
Lechu
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Default [western blotting] Abcam's ab9484 b-actin loading control vs casein blocking



Hi all,

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.

best, Lech

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Old 12-08-2006, 07:57 PM
Pow Joshi
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Default [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.

best
pow



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ab9484 , abcam , bactin , blocking , blotting , casein , control , loading , western


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