BETA ACTIN?????

john_cromwell at my-dejanews.com john_cromwell at my-dejanews.com
Fri Oct 16 14:54:22 EST 1998


Beta-actin (or nuclear histone) is widely accepted in the literature as a
loading control for Western blots. It generally remains the same proportion
of total cell protein except under circumstances that your experimental
conditions are dramatically affecting cytoskeletal rearrangement or cell
adhesion. Without knowing your total protein concentration its the next best
alternative. Unfortunately, it means that you'll have to run several blots
until your loading control is equal in your experimental groups. I don't know
of any articles that review the expression patterns of b-actin, but I hope
this helps.

John W. Cromwell, M.D.
In article <3626717A.FFF1302F at julian.uwo.ca>,
  Dave O'Neill <doneill2 at julian.uwo.ca> wrote:
> HI,
> Due to technical problems (received in sample buffer)  I was unable to
> quantify the total protein content of my cultured P19 (embryonic stem
> cells) cells.  I ran equal volumes on a mini-gel and then stained them
> with commassie blue.  I quantified the entire lane and tried to estimate
> relative protein loads.  Realizing this is a less desirable method, I
> wanted to standardize my protein loads.  I was told I could immunoblot
> with beta-actin.  I'm only concerned about equaling the loads in each
> lane rather than the absolute protein content.
>
> Is Beta Actin always proportion to total cellular protein?  Do you know
> a I might find a basic review on Beta Actin?  Often textbooks fail to
> mention its relative expression to total protein, or its induction??.
> Any thoughts are appreciated.
> Kind regards,
> Dave O'Neill
>
>


--
John W. Cromwell, M.D.

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