c-bsa的制备（The preparation of c - bsa）.docVIP

c-bsa的制备（The preparation of c - bsa） bovine serum albumin Improve the bovine serum albumin (C / BSA) isoelectric point, can induce the in situ immune complex nephritis type, mechanism has been confirmed 1. C BSA and basement membrane by injection into the body of an animal can be planted in the glomerulus (GBM), and with negative charge of GBM deposition reaction induced nephritis 2. In this study, a method for the synthesis of stable bovine serum albumin (BSA) was designed and optimized, and the model of in situ composite nephritis was established. 1 materials and methods 1.1 materials 1.1.1 experimental animals The hybrid male rabbits were weighed (2.2 + 0.3) kg, and the urine protein was qualitative (- to +), which was provided by Tianjin experimental animal center. 1.1.2 C prepared by BSA reagent Bovine serum albumin (BSA), Sigma; two (EDC), carbide imine Sigma; anhydrous ethylenediamine (EDA), AR., Tianjin Chemical Reagent Factory; amphoteric electrolyte pH 4 ~ 10 (Ampholin), Beijing Military Medical Science Academy of the PLA; research Chinese blood dialysis bag, liquid Academy of sciences. 1.1.3 electrophoresis materials and equipment Electrophoresis: DYCP 36 electrophoresis (61 Beijing electronic instrument factory); chymotrypsin (pI=8.1) and cytochrome C (pI=9.8), soybean trypsin inhibitor (pI=4.55), were provided by Sigma company. The other reagents are grade A.R.. 1.2 method 1.2.1 C and BSA preparation process 700 ml ethylenediamine (a water), plus 5 L double distilled water, plus 3.5 L of 6 mol/L hydrochloric acid, pH 4.75, stirring, cooling to 25 DEG C. The addition of 300 ml BSA (stirring in the solution for 60 g BSA) aqueous solution, then add 24 g EDC, 25 DEG C, stirring for 6 hours, then add 300 ml acetic acid buffer solution 4 mol/L pH 4.75, after stopping the reaction, adding the powder of ammonium sulfate saturation reached 0.7, stirring, standing for 8 hours in and collected by centrifugation, dissolved in 500 ml water solution for water desalti