Total protein is an effective loading control for cerebrospinal fluid western blots.

ABSTRACT:

Background

Cerebrospinal fluid (CSF) has been used to identify biomarkers of neurological disease. CSF protein biomarkers identified by high-throughput methods, however, require further validation. While Western blotting (WB) is well-suited to this task, the lack of a validated loading control for CSF WB limits the method's accuracy.

New method

We investigated the use of total protein (TP) as a CSF WB loading control. Using iodine-based reversible membrane staining, we determined the linear range and consistency of the CSF TP signal. We then spiked green fluorescent protein (GFP) into CSF to create defined sample-to-sample differences in GFP levels that were measured by WB before and after TP loading correction. Levels of CSF complement C3 and cystatin C measured by WB with

SUBMITTER: Collins MA 

PROVIDER: S-EPMC4540354 | biostudies-literature | 2015 Aug

REPOSITORIES: biostudies-literature