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kiki06 · #1 (**permalink**) 03-04-2006, 09:16 AM

Hey!, i need some ideas for loading controls...

what do you guys use for loading controls in western blot?

beta-actin?
tubulin?

what is the best to use?

amicia · #2 (**permalink**) 07-22-2006, 09:10 AM

Hello Kiki06,
the answer to your question is depend:
you have to choice which type of control works better. For exemple, in human studies (cell cultures and tissue) I used actin, beacuse is really well express in many cellular compartiments (cytosol, but also mitochondia and for somebody also in nuclei), but also gamma-tubulin generally works very well, but in my knowledge this last one is used only for cytosol protein.
Moreover, if you're evaluating the phosphorilation state of some proteins, I suggest you to use antibody against the phosphorilated state and the non-phosphorilated state, in order to compare the difference between the different treatments in phosphorilated state (of course the amount of no-phopho has to be the same). hope to have help you.
regards

kiki06 · #3 (**permalink**) 07-24-2006, 10:21 PM

thank you Amicia!

I think I will need to check if I can use gamma-tubulin...

shalmoli · #4 (**permalink**) 01-24-2007, 08:28 AM

I think actin will be a good choice

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Signalling is an integral part of the cell neural network. Phosphorylation and other protein modifications modulate this interaction. Post and discuss the study of protein-protein interactions, and signalling biology protocols and science here.