[γ–32P]-ATP was purchased from MP Biomedical (Irvine, CA, USA). Unlabeled deoxynucleoside triphosphate (dNTPs) (ultrapure) were obtained from Pharmacia. Unlabeled dNTPs (ultrapure) were obtained from Pharmacia. Magnesium acetate, magnesium chloride and Trizma base were from Sigma. Urea, acrylamide and bis-acrylamide were from National Diagnostics (Rochester, NY, USA). Oligonucleotides, including those containing 8-oxo-guanine, were synthesized by Operon Technologies (Alameda, CA, USA). All modified nucleotides including N6-MedATP, 6-Cl-PTP, 6-Cl-2APTP, O6-MedGTP, N2-MedGTP, 2-6-dATP, dITP and 8-oxo-dGTP were obtained from Trilink Biotechnologies (San Diego, CA, USA). All non-natural nucleotides including IndTP, 5-MeITP, 5-Et-ITP, 5-EyITP, 5-NITP, 4-NITP and 6-NITP were synthesized and purified as previously reported (23 (link)–26 (link)). All other materials were obtained from commercial sources and were of the highest available quality. Exonuclease-deficient bacteriophage T4 DNA polymerase (gp43exo− (Asp-219 to Ala mutation)) was purified and quantified as previously described (27 (link)). Recombinant human polymerase delta (pol δ) and human polymerase eta (pol η) were purified as previously described (28 (link),29 (link)). All DNA polymerases used in this study were >97% pure as assessed by sodium dodecylsulphate-polyacrylamide denaturing gel electrophoresis.
Acrylamide
Manufactured by National Diagnostics
Sourced in United States
Acrylamide is a chemical compound used as a laboratory reagent. It is a colorless, odorless, crystalline solid that is soluble in water and other polar solvents. Acrylamide is commonly used in the preparation of polyacrylamide gels for electrophoresis, a technique used to separate and analyze biological macromolecules such as proteins and nucleic acids.
Automatically generated - may contain errors
Lab products found in correlation
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Magnesium acetate, by Merck Group (1 mentions)
Urea, by National Diagnostics (1 mentions)
Bis acrylamide, by National Diagnostics (1 mentions)
Magnesium chloride, by Merck Group (1 mentions)
Trizma base, by Merck Group (1 mentions)
γ 32p atp, by MP Biomedicals (1 mentions)
8 oxo dgtp, by TriLink (1 mentions)
Colchicine, by Merck Group (1 mentions)
Pf 573228, by R&D Systems (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
Enhanced chemiluminescence, by Thermo Fisher Scientific (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Fasudil, by Cayman Chemical (1 mentions)
Paxillin, by Abcam (1 mentions)
Fak c 20, by Santa Cruz Biotechnology (1 mentions)
Yap1 d8h1x, by Cell Signaling Technology (1 mentions)
Glass bottom imaging dishes, by Cellvis (1 mentions)
Myosin light chain 2, by Cell Signaling Technology (1 mentions)
Phalloidins, by Thermo Fisher Scientific (1 mentions)
4 protocols using acrylamide
1
Synthesis and Purification of Modified Nucleotides
2
Signaling Inhibitors and Activators
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The following inhibitors or activators were used for signaling studies: 1–5 mM EDTA, 10 µM PP2, 1 µM colchicine (Sigma), 6 mM acrylamide (National Diagnostics, Atlanta, GA, USA), 1 µM AG82 FAK/Src inhibitor, 3 µM manumycin (Calbiochem, San Diego, CA, USA), 500nM PF 573228 (R&D Systems, MN, USA), and 5 µM LPA (Enzo Life Sciences).
3
Western Blot Protein Separation and Detection
4
Immunofluorescence Assay for Cell Adhesion
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Antibodies (and their source) against the following targets were used: fibronectin (BD Biosciences, #610077); paxillin (Abcam, ab32084); phospho-myosin light chain 2 (pThr18/pSer19; Cell Signaling Technology, #3674); myosin light chain 2 (Cell Signaling Technology, #3672); vinculin (hVin-1; Millipore-Sigma V9131); phospho-FAK (pTyr397; Cell Signaling Technology, #3283); FAK (C-20; Santa Cruz Biotechnology; sc-558); YAP1 (D8H1X; Cell Signaling Technology, #14074). DAPI (4',6-diamidino-2-phenylindole dihydrochloride), Hoechst 33342, and phalloidins conjugated to Alexa Fluor 488, 594, or 647 were from ThermoFisher Scientific. Fasudil (also known as HA-1077) was from Cayman Chemical, PF-562271 was from ChemScene, and Blebbistatin (#203389) and latrunculin A (L5163) were from Millipore Sigma. Acrylamide and N,N-methylenebisAcrylamide were purchased from National Diagnostics. Human plasma-derived fibronectin (Corning #356008) was from Thermo Fisher. Glass-bottom imaging dishes were from Cellvis. Other chemicals and reagents, unless otherwise noted, were purchased from Millipore Sigma.
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