A BN-PAGE-based method for histochemical staining and quantification of DLDH diaphorase activity is presented. The method involves the resolution of mitochondrial proteins by BN-PAGE that is capable of localizing a DLDH protein band. Diaphorase activity staining is then achieved by DLDH-mediated formation of NBT formazan at the expense of NADH. The technique should be the method of choice when DLDH dehydrogenase activity can not be measured spectrophotometrically due to the presence of interfering chemicals or reactive reagents in assay mixtures. Moreover, the method can also be used for isolation of mitochondrial DLDH that is to be analyzed by other techniques such as mass spectrometry in studying DLDH post-translational modifications.
|Title of host publication||2007 AIChE Annual Meeting|
|State||Published - 1 Dec 2007|
|Event||2007 AIChE Annual Meeting - Salt Lake City, UT, United States|
Duration: 4 Nov 2007 → 9 Nov 2007
|Other||2007 AIChE Annual Meeting|
|City||Salt Lake City, UT|
|Period||4/11/07 → 9/11/07|