The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ELISA: Use at an assay dependent dilution.
With this antibody, we have found that blocking with 5% goat or donkey serum significantly reduces background as compared to BSA or milk.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Serine/threonine protein kinases phosphorylate the OH group of serine or threonine (which have similar sidechains).
Protein kinase C is actually a family of protein kinases consisting of ~10 isozymes. They are divided into three subfamilies: conventional (or classical), novel, and atypical based on their second messenger requirements.
While serine/threonine kinases all phosphorylate serine or threonine residues in their substrates, they select specific residues to phosphorylate on the basis of residues that flank the phosphoacceptor site, which together comprise the consensus sequence. Since the consensus sequence residues of the substrate to be phosphorylated make contact with the catalytic cleft of the kinase at several key amino acids (usually through hydrophobic forces and ionic bonds), a kinase is usually not specific to a single substrate, but instead can phosphorylate a whole "substrate family" having common recognition sequences.
The consensus sequence of protein kinase C enzymes is similar to that of protein kinase A, since it contains basic amino acids close to the Ser/Thr to be phosphorylated. Their substrates are MARCKS proteins, MAP kinase, transcription factor inhibitor IkappaB, the vitamin D3 receptor VDR, Raf kinase, calpain, and the epidermal growth factor receptor.