Improved methods for total and chloroplast protein extraction from Cajanus species for two-dimensional gel electrophoresis and mass spectrometry
Fig 5
SDS-PAGE and immunoblot analysis of the proteins to determine the purity of extracted chloroplast protein.
(A) One dimensional SDS-PAGE analysis of total and chloroplast proteins from pigeon pea genotype C. cajan and C. scarabaeoides showing absence of significant number of cytosolic proteins in chloroplast protein fraction in comparison to total protein fraction. (B) Immunoblot analysis of chloroplast and total protein, using LHCb1/LHCⅡ type I chlorophyll a/b binding protein antibody, showing higher band intensity in the proteins extracted from chloroplast fraction in comparison to the one from total proteins. Equal loading of the protein was confirmed with ponceau staining and anti- LHCb1/LHCⅡ type I was used at a concentration of 1:2500.