ABSTRACT
OBJECTIVE: To optimize the two-dimensional electrophoresis (2-DE) method for the proteome analysis of the Cervus nippon antler, and compare the protein maps of different parts of Cervus nippon antler. METHODS: The total proteins of Cervus nippon antler were extracted by protein lysate containing 7 mol/L Urea, 4% CHAPS, 2 mol/L Thiourea, 65 mmol/L DTT, 1 mmol/L PMSF and 0.2% Bio-Lyte. The proteins were separated by immobilized pH 3 - 10 linear gradient (IPG), 7 cm strips as the first dimension. Isoelectric focusing conditions were optimized. 12% SDS-PAGE was used as the second dimension electrophoresis. The gels were stained with Coomassie brilliant blue and analyzed by PDQuest analysis software. RESULTS: The contents of total protein and the numbers of protein points of three different parts of Cervus nippon antler reduced gradually from the top to the bottom. Comparing three maps of different parts of Cervus nippon antler, there were 18 different protein points. Isoelectric point, molecular weight and gray value of each different protein point were calculated. CONCLUSION: An optimized two-dimensional electrophoresis method for the proteome analysis of the Cervus nippon antler is established. The 2-DE profiles of different parts of Cervus nippon antler exist obvious differences. The different protein points can be used as reference for Cervus nippon antler quality control and evaluation.
Subject(s)
Antlers/chemistry , Deer , Proteins/chemistry , Proteome/analysis , Animals , Electrophoresis, Gel, Two-Dimensional , Electrophoresis, Polyacrylamide Gel , Hydrogen-Ion Concentration , Isoelectric Focusing , Male , Materia Medica/chemistry , Molecular Weight , Proteins/isolation & purification , Quality Control , SoftwareABSTRACT
OBJECTIVE: To optimize the formulation of Eisemia foetida protein (EFP) burn spray. METHODS: A five-factor, three-level response surface method was employed; The response variable was the proliferation effect of EFP on NIH3T3 cells. RESULTS: The optimization formulation was as follows: the proportion of EFP, glycerol and mannitol was 0.91%, 1.42% and 5%, respectively; 0.02 mol/L Na2 HPO4 and 0.01 mol/L citric acid buffer system corresponding pH value was 7.0. CONCLUSION: The response surface method is reliable, efficient and suitable for optimizing the formulation of EFP burn spray.
Subject(s)
Cell Proliferation/drug effects , Chemistry, Pharmaceutical/methods , Mannitol/chemistry , Materia Medica/chemistry , Oligochaeta/chemistry , Proteins/chemistry , Aerosols , Animals , Burns/drug therapy , Citric Acid/chemistry , Hydrogen-Ion Concentration , Materia Medica/pharmacology , Mice , NIH 3T3 Cells , Preservatives, Pharmaceutical/chemistryABSTRACT
An in vitro detection method of the gastrointestinal absorption of Pilose Antler protein was established for mixed protein activity. Five bands of protein with molecular weight of 17.8-160 kD derived from the Pilose Antler were extracted and sufficiently labeled with FITC (FITC-PE). The stability and variation of FITC-PE in gastrointestinal circumstances were detected by native polyacrylamide gel electrophoresis and confocal laser scanning microscope. Results showed that the main component of FITC-PE kept invariant after being reacted with artificial gastric fluid and artificial intestinal fluid. The fluorescence signal was detected 20 min after administration in the valgus intestinal purse experiment, and three kinds of protein, with molecular weight of 45, 25, and 17.8 kD, were detected in the mixture of absorbent protein. The research laid the foundation for the further in vivo study of Pilose Antler protein. Meanwhile, it would be an in vitro screening method for the absorption, distribution and metabolism of mixed protein from traditional Chinese medicine.
Subject(s)
Antlers/chemistry , Deer , Intestinal Mucosa/metabolism , Materia Medica/pharmacokinetics , Proteins/pharmacokinetics , Animals , Fluorescein-5-isothiocyanate , Gastric Mucosa/metabolism , Intestinal Absorption , Male , Materia Medica/chemistry , Materia Medica/isolation & purification , Microscopy, Confocal , Molecular Weight , Native Polyacrylamide Gel Electrophoresis , Proteins/chemistry , Proteins/isolation & purification , Rats , Rats, WistarABSTRACT
OBJECTIVE: To offer scientific basis of distinguish p recious animal TCM. METHOD: Molecular weights of the proteins were measured by modified sodium dodecyl sulphate-polyaerylamide gel eleetrophoresis (SDS-PAGE). RESULTS: The bands were always quite distinct. CONCLUSION: The method is accurate, reliable and with good reproducibility. It can supply reference datas on quality con-tol and preparation produce of snakes, Calculus Bovis and Cervus.
Subject(s)
Electrophoresis, Polyacrylamide Gel/methods , Materia Medica/chemistry , Medicine, Chinese Traditional , Proteins/chemistry , Animals , Cattle , Deer , Drug Combinations , Molecular Weight , Proteins/isolation & purification , Proteins/standards , Quality Control , Reproducibility of Results , Snakes , Sodium Dodecyl SulfateABSTRACT
The comparative study of the proteins of Bungarus multicinctus multicinctus and Natrix annularis was carried out by SDS-PAGE. The results showed that there were significant differences between their SDS-PAGE figures. On the basis of the study the molecular weights of their characteristic proteins were determined. The results could be regarded as a reference for identification of Bungarus multicinctus multicinctus and its confused species.