Phosphorylation of Cx43 was quantified by immunoprecipitation with an anti-Cx43 antibody followed by immunoblot analysis using an anti-phosphoserine antibody.
The expression levels of MerE protein were measured in transgenic plants (lines E2) by immunoblot analysis using a polyclonal anti-MerE antibody, which was prepared in a previous study (Kiyono et al. 2009).
A 30 kDa polypeptide detected by immunoblot analysis using anti-phosphotyrosine antibody in livers from rats 48 to 72 h after administration of a single dose of CCl4 was identified as galectin-3 induced in cytoplasm of periportal hepatocytes and phosphorylated on tyrosine residue(s).
ATF-2 phosphorylation was detected by immunoblot analysis using a phospho-ATF-2 (Thr71) antibody.
The electrophoretic pattern of some proteins was investigated by immunoblot analysis using specific polyclonal antibodies.
Expression of each protein was confirmed by immunoblot analysis using anti-His antiserum (Fig. 3A).
Reactions were stopped by addition of Laemmli buffer and analyzed by immunoblot analysis using anti-V5 antibody.
We also confirmed the mutation by immunoblot analysis using anti-phospho-APP Thr668 specific antibody (Fig. 1D).
Next, we examined protein levels of APP C-terminal fragments (CTFs) by immunoblot analysis using anti-APP cytoplasmic tail antibodies (Fig. 4).
To verify the role of p53 in DOX-induced autophagic cell death at a biochemical level, the conversion of LC3-I to LC3-II, an indicator of autophagy activation, was assessed by immunoblot analysis using LC3 antibody (Figure 3A).
The presence of O-linked glycosylation (O-GlcNAc) was determined by immunoblot analysis using the O-GlcNAc Western Blot Detection system from Pierce (Rockford, IL), and according to the directions supplied by the manufacture.
