Methods for producing a protein extract from cells, such as cells containing viral proteins, are provided. In general terms, the methods involve: increasing the pH of the cells to a pH of at least about pH 10.0 to produce an intermediate composition, and then, in the presence of a non-...
Protein extraction from tissues and cultured cells is the first step for many biochemical and analytical techniques (PAGE, Western blotting, mass spectrometry, etc.) or protein purification. Efficient disruption and homogenization of animal tissues and cultured cells are required to ensure high yield...
Protein extraction from cultured cells Approximately 1–5 g (FW) of Cj156-S and CjY cells were inoculated in 20 or 100 ml of culture medium and cultured for 1–2 weeks. The grown cells were collected and ground in liquid nitrogen. An equal volume of extraction buffer [50 mM Tris...
in theory, are prone to stationary phase regulatory mechanisms such as ribosome breakdown. Because this contradicts current opinions, we performed a thorough characterization of the cell-free extract generated from resting cells. Since ribosomes are the main parameter in cell-free translation systems, ...
Miss Sui-QiCheng, ...Tian-MiaoOu, inMethods, 2019 1Introduction All cellular proteins are produced from the mRNA in the process of translation, in which functional ribosomes andprotein complexesare involved by sequentially assembling onto the mRNA strands. The orderly and controllable develo...
The SOD protein standard was prepared by extraction from E. coli cells with Marathon A2 microparticles. Cells were incubated for 2 h at ratio 0.7 v/v (resin:cells) in 50 mM TRIS buffer at pH 8.0. Protein was desorbed with 0.2 M NaCl and subsequently purified by gel filtration (Superdex...
Total protein quantification methods Protein concentration quantitation is an integral part of any laboratory workflow involving protein extraction, purification, labeling or analysis. Pierce Protein Assays provide a wide range of options for a...
The off-AID system is a powerful method for depletion of specific proteins within fission yeast cells. Methods Strains and media Schizosaccharomyces pombe strains used in this study are listed in Table 2. Constructions of strains HM1813, HM1905, HM2473, HM2475, HM2491, HM2550, HM2551, HM...
Elucidation of both the three-dimensional structure and the dynamics of a protein is essential to understand its function. Technical breakthroughs in single-particle analysis based on cryo-electron microscopy (cryo-EM) have enabled the three-dimensional
Cells were solubilized with lysis buffer (8 M urea and 4% (w/v) SDS in 50 mM Tris–HCl, pH 7.5) and incubated at 95 °C for protein extraction. Cell lysates were sonicated on ice using the Bioruptor UCD-250T (Cosmo Bio, Tokyo, Japan). The solution was subjected to chlorofor...