Absolute quantitation of endogenous proteins with precision and accuracy using a capillary Western system.

ABSTRACT: Precise and accurate quantification of protein expression levels in a complex biological setting is challenging. Here, we describe a method for absolute quantitation of endogenous proteins in cell lysates using an automated capillary immunoassay system, the size-based Simple Western system (recently developed by ProteinSimple). The method was able to accurately measure the absolute amounts of target proteins at picogram or sub-picogram levels per nanogram of cell lysates. The measurements were independent of the cell matrix or the cell lysis buffer and were not affected by different antibody affinities for their specific epitopes. We then applied this method to quantitate absolute levels of expression of protein kinase C (PKC) isoforms in LNCaP and U937 cells, two cell lines used extensive

SUBMITTER: Chen JQ 

PROVIDER: S-EPMC3805113 | biostudies-literature | 2013 Nov

REPOSITORIES: biostudies-literature