生化分离（Biochemical separation）.docVIP

生化分离（Biochemical separation） 1, RCF is the relative centrifugal force value of RCF (g) =1.1 * 10-5 (RPM) 2 * r RPM: RPM rpm/60=N; R to MM calculation, sometimes with a separation factor of Kc, defined as Fc type centrifugal force by unit mass of the material, the separation factor is a parameter to measure the degree of centrifugal the representation can achieve centrifugal acceleration and acceleration of gravity ratio in the literature often how much G is represented by the centrifugal force. 2, cell lysis buffer: if the object is a product of intracellular biochemical separation, solid-liquid separation of fermentation bacteria precipitation is collected after some broken cell means that the intracellular product release, before the cells were broken, with broken cell buffer was suspended, the typical cell lysis buffer general 0.1-0.2mol/L, pH phosphate buffer solution or Tris buffer 7 - 8 (similar to the intracellular environment); some may also contain antioxidants, such as two 4-dithiothreitol DTT, 2- Cys, 2-BME mercaptoethanol and cysteine glutathione GSH (reduced), the activity of the protein thiol to protect; may also add some targeted inhibitors such as serine protease inhibitor PMSF, cysteine proteinase inhibitor iodoacetic acid, metalloproteinase EDTA. 3, Critical micelle concentration: critical micelle concentration, said (anti) the lowest concentration required for surfactant micelle formation, CMC said, this is the system characteristics, and surfactant chemical structure, solvent, temperature and pressure and other factors, namely: 4 Inclusion body inclusion bodies, the current reorganization protein has thousands of species, which accounted for a large part of the recombinant protein is expressed in the host bacteria E.coli. The recombinant protein expressed in Escherichia coli often deposited in inclusion bodies in the cytoplasm, showed no activity of insoluble aggregates, i.e. the primary structure is correct, without proper folding and showed no