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). WebJan 1, 2013 · 3.2 Preparation of Nuclear and Cytosolic (S100) Extracts Below is the general procedure we use to prepare extracts from 12 L suspension cultures of HeLa S3 cells expressing FLAG-epitope tagged Mediator subunits. Additional protocols are provided elsewhere ( 9 – 11 ).
Preparation of Hela Cell Nuclear and Cytosolic S100 Extracts for In
WebPreparation of HeLa cell nuclear and cytosolic S100 extracts for in vitro splicing. Methods Mol Biol. 1999;118:309-14.doi: 10.1385/1-59259-676-2:309. Authors. A Mayeda 1 , A R Krainer. Affiliation. 1Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, Florida, USA. WebS100 protein (pS100) is a commonly used melanocytic antigen because of its high sensitivity in spite of relatively low specificity. SRY-related HMG-box 10 protein (SOX10) is a transcription factor characterising neural crest-derived cells. It is uniformly expressed mostly in the nuclei of melanocytes, neural, and myoepithelial cells. spring session timeout redirect
Preparation of Hela Cell Nuclear and Cytosolic S100 …
WebMar 17, 2000 · To this end initiation complexes were formed in the presence of the SV40 T-Ag, dialyzed cytosolic S100 replication extract and ATP/Mg 2+ using protein-free DNA and chromatin reconstituted with control and cross-linked octamers (histone:DNA weight ratio 1:1.2) as template. 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 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). sheraton montazah hotel alexandria egypt