Abstract
The first experiments were carried out to examine the conditions for the preservation of saliva protein, the effect of the use or non-use of preservatives and procedures such as condensation of the sample, freezing and thawing, and lyophilization upon the electrophoretic picture on polyacrylamide gel disc was studied. The changes of the electrophoretic picture depending on the condition of preservation were studied at each temperature (-80-+50°C) with the passage of time.
As to each electrophoretic pattern, the presence or absence of amylase activity was studied, and attempts were made to detect amylase protein by immunoelectrophoresis. Rocket immunophoresis of each protein in the sample was carried out against various antibodies to serum protein and the differentiation with immunodiffusion was attempted. Each electrophoretic zone was subjected to SDS electrophoresis to estimate the molecular weight. Isoelectric focusing was performed to estimate the isoelectric point. Conditions for the appearance of abnormal zone were also studied. The results obtained are as follows :
1) Addition of EDTA was useful for the preparation and preservation of the whole saliva sample.
2) Under freezing, all the samples were essentially stable for several months. Since appearing and disappearing zones were noted in some parts of all the samples, it appears that care must be taken with preservation under freezing.
3) When samples were not frozen, the PVP-condensed sample with the addition of EDTA was most stable.
4) When the whole saliva was treated with urea or guanidine, considerable changes were noted in the regions II and III.
5) The molecular weight of protein in the whole saliva ranged from approximately 20, 000 to 115, 000. In isoelectric focusing, most of the patterns in the whole saliva were found to be on the acidic side. The main components of the whole saliva appear to be acidic protein.
6) Saliva protein related to serum protein such as Alb, Pre Alb, IgA, IgG, Tf and Secretory Piece was identified.
