Cytosolic s100 extracts
While waiting for one of the centrifugation steps in Subheading 3.2. , the supernatant obtained in Subheading 3.2.6. can be processed further to obtain an S100 extract, which contains cytosolic components as well as many, but not all, nuclear components, which are extracted in the hypotonic buffer A. 1. 1. … See more The cells are grown in spinner flasks at 37°C. The cell density should be maintained daily between 2×105 and 5×105. The doubling … See more WebMayeda, A., Krainer, A. R. (1999) Preparation of HeLa cell nuclear and cytosolic S100 extracts for in vitro splicing. Methods in Molecular Biology, 118. pp. 309-14. ISSN 1064-3745 (Print)1064-3745
Cytosolic s100 extracts
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WebAug 16, 1999 · Total proteins (∼5 μg) from HeLa cell nuclear and cytosolic S100 extracts were probed with rabbit anti-RNPS1 antibody. Purified baculovirus r-RNPS1 (∼5 pmol) was included as a control. The estimated abundance of RNPS1 in the nuclear and cytosolic extracts is ∼0.3 and ∼0.01 pmol/μg protein, respectively. WebJun 15, 1998 · Preparation of cytosolic S100 replication extracts and the SV40 T-Ag. Cytosolic S100 extracts were prepared from HeLa S3 cells exactly as described (Stillman and Gluzman, 1985). SV40 T-Ag was …
WebMayeda, A., Krainer, A. R. (1999) Preparation of HeLa cell nuclear and cytosolic S100 extracts for in vitro splicing. Methods in Molecular Biology, 118. pp. 309-14. ISSN 1064-3745 (Print)1064-3745 URL: http://www.ncbi.nlm.nih.gov/pubmed/10549533 DOI: 10.1385/1-59259-676-2:309 Abstract WebThe protocol described is for preparation of both nuclear and cytoplasmic S100 extracts from HeLa cells in an analytical and small-to-medium preparative scale. The nuclear extracts are active in splicing of many different pre-mRNAs, while S100 extracts are not.
WebMar 18, 1997 · Preparation of cytosolic extracts (S100) was as previously described . Electrophoretic Mobility-Shift Assays (EMSAs). RNA oligonucleotides were synthesized by the University of Pennsylvania Nucleic Acid Core Facility and were 5′-end labeled using T4 polynucleotide kinase (NEB, Beverly, MA) and [γ-32 P]ATP (Amersham). WebPreparation of Hela Cell Nuclear and Cytosolic S100 Extracts for In Vitro Splicing Springer Nature Experiments Following the initial discovery of split genes in 1977, it took several years before in vitro systems were successfully developed to study the biochemistry of pre-mRNA ...
WebFeb 5, 2016 · The cytosolic marker Pgk1 was recovered from S100 fraction . The fact that ubiquitinated Ste6* is distributed in P20 and P100 …
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