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In biochemistry, phenylmethylsulfonyl fluoride (PMSF) is a serine protease inhibitor (serine hydrolase inactivator) commonly used in the preparation of cell lysates. PMSF does not inactivate all serine proteases. [1] The effective concentration of PMSF is between 0.1 - 1 mM.
By developing specific and controllable bioorthogonal reactions, scientists have opened up the possibility of hitting particular targets in complex cell lysates. Recently, scientists have adapted click chemistry for use in live cells, for example using small molecule probes that find and attach to their targets by click reactions.
Plasmolysis is the contraction of cells within plants due to the loss of water through osmosis. In a hypertonic environment, the cell membrane peels off the cell wall and the vacuole collapses. These cells will eventually wilt and die unless the flow of water caused by osmosis can stop the contraction of the cell membrane .
Radioimmunoprecipitation assay buffer (RIPA buffer) is a lysis buffer used to lyse cells and tissue for the radio immunoprecipitation assay (RIPA). [1] [2] This buffer is more denaturing than NP-40 or Triton X-100 because it contains the ionic detergents SDS and sodium deoxycholate as active constituents and is particularly useful for disruption of nuclear membranes in the preparation of ...
Cell lysis is a critical step in the purification of enzymes from bacterial cells, various components are commonly included in lysing buffers to facilitate effective cell disruption and release of the target enzyme. These components include detergents, salts, and enzymes, each playing a specific role in the lysis process.
In most cases, preclearing the lysate at the start of each immunoprecipitation experiment (see step 2 in the "protocol" section below) [6] is a way to remove potentially reactive components from the cell lysate prior to the immunoprecipitation to prevent the non-specific binding of these components to the IP beads or antibody.
The concept of "protein-sparing modified fast" (PSMF) was described by George Blackburn in the early 1970s as an intensive weight-loss diet designed to mitigate the harms associated with protein-calorie malnutrition [8] and nitrogen losses induced by either acute illness or hypocaloric diets in patients with obesity, in order to adapt the patient's metabolism sufficiently to use endogenous fat ...
The steps of alkaline lysis can be summarized as the formation of a pellet, resuspension of the pellet in solution, cell lysis, neutralization, and centrifugation. [ 2 ] Alkaline lysis takes advantage of the small and supercoiled physical composition of plasmid DNA compared to chromosomal DNA, along with its ability to reanneal double stranded ...
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