Studies On Detection And Distribution Of Plcln Protein

pICln protein was cloned in 1992 by Paulmichl from MDCK(madin-darby canine kidney cell line).As a new protein that can regulates cell volume , some researchers believe that it is a chloride channel protein, while others suggest it is a cytoplasmic regulators.We have done some researches on it.The results are as follows:一.Production of antibody against C-end eighteen peptide fragment of pICln proteinA synthesized peptide with 19 amino acid(NH2-EVDTTPTVAGQFEDADVDH-COOH) residues of pICln C-terminal from MDCK cells line was used to immune rabbit,the antibody against the peptide was produced.The results confirmed that the peptide antibody was specific to pICln protein by ELISA.二.Assay of pICln in tissue and cells with Western BlotWe detected some tissues and cells with Western Blot,found that they all had pICln expression and 40kDa immunoreactive bands were detected.三.Distributions of pICln in LLC-PK1Distributions of pICln in cells of LLC-PK1 were detected by immunohistochemical. pICln of LLC-PK1 located mainly in microsome and mitochondria of insoluble fraction.Compared to isotonic solution,the distribution of pICln in LLC-PK1 cells had a obvious change in hypotonic solution,which show the sensitivity of pICln to hypotonic solution.