Lysis efficiency

The addition of water to a crude P2 fraction changes the osmotic pressure of the medium. The synaptosomes will easily lyse as a result of this osmotic shock, whereas the mitochondria and vesicles are largely unaffected. The lysis efficiency of the synaptosomes determines the yield of the synaptosomal plasma membrane preparation. 

One advantage of the technique is it is internally referenced. That is, variations in cell lysis efficiency will not have an effect because what is measured is the amount of plasmid DNA relative to a genomic gene in a given sample. However, differences in the efficiency of copying can have an effect. 

As seen in the genetic map, the genes after gene 1.1, transcribed by the T7 RNA polymerase, code for proteins that are involved in T7 DNA synthesis, the formation of virus coat proteins, and assembly. Three classes of T7 proteins are formed class I, made 4-8 minutes after infection, which use the cell RNA polymerase class II, made 6-15 minutes after infection, which are made from T7 RNA polymerase and are involved in DNA metabolism class III, made from 6 minutes to lysis, which are transcribed by T7 RNA polymerase and which code for phage assembly and coat protein. This sort of sequential pattern, commonly seen in many large double-stranded DNA phages, results in an efficient channeling of host resources, first toward DNA metabolism and replication, then on to formation of virus particles and release of virus by cell lysis. 

The addition of Tween 20 to the buffers Is necessary to enable optimal collection of the magnetic beads on the sides of the MTP wells and also to facilitate efficient cell lysis In Step 1. Bead volumes etc. are based on the QIAgen protocol and may require adjustment If reagents from another supplier are used. 

Gruber, M Schodin, B A., Wilson, E. R., and Kranz, D. M. (1994) Efficient tumor cell lysis mediated by a bispecific single-chain antibody expressed in Escherichia colt J Immunol. 152, 5368—5374. 

The observed enhancement of the catalytical efficiency of immobilized trypsin in the presence of immobilized heparin was shown to be due to the association of the products of fibrinolysis (as in the case of fibrinogen) which removed them out of reach of immobilized trypsin 137>. In fact, as is seen from Fig. 9, the lysis rate gradually decreases as the reaction products are accumulated, i.e., accumulation of the lysis products causes the poisoning of the catalyst. Addition of new polymer, but this time of that containing immobilized heparin only, again leads to an increase of the lysis rate (Fig. 10). 

Viruses are complex particles, entering the cells by fusion of their envelope to the plasma membrane or by endocytosis followed by the escape of the capsid by membrane fusion or lysis (Sodeik, 2000). The diameter of the viral particle could be several hundred nanometers, implying a very inefficient diffusional movement in the cytoplasm, based on those physicochemical considerations that were discussed above (Kasamatsu and Nakanishi, 1998). Despite these limitations, those viruses that replicate in the nucleus have evolved sophisticated mechanisms to ensure a highly efficient nuclear delivery of their genetic material. Since these mechanisms may provide a conceptual framework to design novel non-viral delivery systems, we shall review some of the key elements that account for the nuclear targeting of certain viruses. 

Another effector function/mode of action of mAbs is the so-called complement-dependent cytotoxicity or complement-mediated cell death . Activation of the complement system can lead to lysis of the antigen-presenting cell, or can induce inflammation reactions aimed at eliminating these cells efficiently. The successive steps of the complement activation can be summarized in a simplified way [9] ... 

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