2 replies to this topic

[dzasp]

Hi everyone,

I'm researching a secreted protein in culture medium from cell lines of prostate cancer. I run a WB without problems, however I need a secreted protein to normalize the results and loading control (like actin in protein extract).

Are there any protein that always be released to culture medium?

Thank you!
Valentina

Edited by dzasp, 29 October 2009 - 09:39 AM.

[bellm]

I have read about others using erk (extracellular signal-regulated kinase) as such a loading control.  
See Coban 2005, J. Leukocyte Biology 78:647-655.  Also see Dittmer 2006, Electrophoresis 27, 2844-2845.

Cheers,
Matt

[Feelcontraire]

dzasp, on Oct 29 2009, 06:03 AM, said:

Hi everyone,

I'm researching a secreted protein in culture medium from cell lines of prostate cancer. I run a WB without problems, however I need a secreted protein to normalize the results and loading control (like actin in protein extract).

Are there any protein that always be released to culture medium?

Thank you!
Valentina

Loading controls are fake-prone.

I would recommend you simply performing ponceau stain, scan and lane densitometry.