All of the chemicals used were of analytical grade. The functional peptides FP-1 and FP-2, and their peptide conjugates, Fluorescein-5-isothiocyanate (FITC), FP-1-FITC and FP-2-FITC were synthesized by Guoping Pharmaceuticals (Hefei, China) with 98% purity. 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP) was purchased from Tokyo Chemical Industry (Tokyo, Japan). Cell Counting Kit (CCK8) was purchased from Solar bio (Beijing, China). Phosphatidylcholine DPPC (1, 2-dihexadecanoyl-sn-glycero-3-phosphocholine) was purchased from Avanti Polar Lipids Co., Ltd (Birmingham, AL, USA). 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) and cell-membrane orange-red fluorescent probe (DIL) staining solution were purchased from Beyotime (Shanghai, China). The mouse fibroblast cell line (L929) was obtained from normal loose subcutaneous connective tissue of mice, and the cell line L was cloned. The L929 cell line was provided by Suzhou Bena Tronlink Biotechnology Co., Ltd (Suzhou, China).
1 1 1 3 3 3 hexafluoro 2 propanol hfip
Manufactured by Tokyo Chemical Industry
Sourced in United States, Japan
1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) is a specialty chemical used in various industrial and research applications. It is a colorless, volatile, and highly polar organic compound with a high boiling point. HFIP is primarily utilized as a solvent, particularly for the dissolution and processing of certain polymers and biomolecules.
Automatically generated - may contain errors
Lab products found in correlation
Unmodified oligodeoxyribonucleotides odns, by Integrated DNA Technologies (4 mentions)
γ 32p atp, by PerkinElmer (3 mentions)
Shrimp alkaline phosphatase, by Thermo Fisher Scientific (2 mentions)
2 4 amidinophenyl 6 indolecarbamidine dihydrochloride dapi, by Beyotime (1 mentions)
Cell counting kit 8, by Solarbio (1 mentions)
17 aag, by Merck Group (1 mentions)
Penicillin, by Nacalai Tesque (1 mentions)
Streptomycin, by Nacalai Tesque (1 mentions)
Iron 3 oxide nanopowder, by Merck Group (1 mentions)
Mcf 7 human breast cancer cells, by American Type Culture Collection (1 mentions)
Formic acid, by Nacalai Tesque (1 mentions)
Thioflavin t, by Merck Group (1 mentions)
Acetonitrile mecn, by Nacalai Tesque (1 mentions)
Sequencing grade modified trypsin, by Promega (1 mentions)
Dipyridamole, by Tokyo Chemical Industry (1 mentions)
Methanol, by Samchun (1 mentions)
Chloroform, by Samchun (1 mentions)
8 protocols using 1 1 1 3 3 3 hexafluoro 2 propanol hfip
1
Functional Peptide Conjugates for Cell Analysis
2
Fabrication of Doxorubicin-Loaded PCL Nanoparticles
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Poly(ε-caprolactone) (PCL, Mw = 80 kDa) (>99.0%), 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) (>99.0%), and DOX (>95.0%) were purchased from Tokyo Chemical Industry Co., Ltd. (TYO, Japan). Iron (III) oxide nanopowder (less than 50 nm particle size) and 17AAG (≥98%) were obtained from Sigma-Aldrich Japan (TYO, Japan). Modified Eagle’s Medium (MEM), trypsin, penicillin, and streptomycin were obtained from Nacalai Tesque, Inc. (Kyoto, Japan). Fetal bovine serum (FBS) was purchased from Tocris Bioscience Inc. (Minneapolis, MN, USA). Alamar blue reagent was obtained from TREK Diagnostics (Cleveland, OH, USA). MCF-7 human breast cancer cells were purchased from the American Type Culture Collection (Manassas, VA, USA).
3
Amyloid-beta Peptide Degradation Assay
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Specific reagents and materials were purchased as follows: Aβ40 (human, DAEFRHDSGY EVHHQKLVFF AEDVGSNKGA IIGLMVGGVV) and Aβ16 (human, DAEFRHDSGY EVHHQK) (Anaspec, Inc., Fremont, CA, USA); NEP (human recombinant, solution in Tris, NaCl and ZnCl2) (R&D Systems, Inc., Minneapolis, MN, USA); IDE (human recombinant, solution in Tris and NaCl) (Bon Opus Biosciences, LLC., Millburn, NJ, USA); sequencing grade modified trypsin (Promega Co., Madison, WI, USA); acetonitrile (MeCN), formic acid (FA), and tetrahydrofuran (THF) (Nacalai Tesque, Inc., Kyoto, Japan); thioflavin T (2-[4-(dimethylamino)phenyl]-3,6-dimethyl-1,3-benzothiazol-3-ium chloride, ThT) (Sigma-Aldrich, Inc., St. Louis, MO, USA); 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) (Tokyo Chemical Industry Co., Ltd., Tokyo, Japan); OptiPlate-384, White Opaque 384-well MicroPlate (384 well plate) (PerkinElmer, Waltham, MA, USA); AMPLIseal (plate seal) (Greiner Bio-One, Baden-Württemberg, Germany); and Protein LoBind® tubes, 0.5 mL (Eppendorf, Hamburg, Germany). All other general chemicals, vials, and gases were of the highest grade available and were obtained from local providers.
4
Biodegradable PCL Scaffold Synthesis
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Both PCL (Mn = 80,000 g/mol) and PDO pellets were purchased from Sigma Aldrich (USA), while 1, 1, 1, 3, 3, 3-hexafluoro-2-propanol (HFIP) and dipyridamole were purchased from Tokyo Chemical Industry Co., Ltd. (Japan). Chloroform, methanol, and ethyl alcohol were purchased from Samchun Chemical (Korea). Medical-grade biodegradable PCL granules were purchased from ROKIT (South Korea). All animal studies were performed in accordance with the ethical guidelines of Chonnam National University Hospital and Medical School (South Korea). All protocols were approved by the Chonnam National University Animal Ethics Committee.
5
Synthesis and Characterization of 5hmU-containing Oligonucleotide
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Unmodified oligodeoxyribonucleotides (ODNs) used in this study were purchased from Integrated DNA Technologies (Coralville, IA, USA). [γ-32P]ATP was obtained from Perkin Elmer (Piscataway, NJ, USA). Shrimp alkaline phosphatase was obtained from the USB Corporation (Cleveland, OH, USA). T4 phage β-glucosyltransferase (T4 β-GT) and all other enzymes unless otherwise specified were purchased from New England BioLabs (NEB). 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP) was purchased from TCI America (Portland, OR, USA). Chemicals unless otherwise noted were obtained from Sigma-Aldrich (St. Louis, MO, USA).
The 5hmU-containing ODN (5′-ATGGCG5hmU GCTAT-3′) was synthesized following previously published procedures [17] (link), and the identity of the modified ODN was confirmed by electrospray ionization-mass spectrometry (ESI-MS) and tandem MS (MS/MS) analyses [18] (link). We chose this particular sequence context because we previously conducted replication studies for a number of DNA lesions in the same sequence context [19] (link)–[21] (link).
The 5hmU-containing ODN (5′-ATGGCG
6
Characterization of O4-alkyldT Lesions
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All chemicals, if not specifically described, were from Sigma-Aldrich (St. Louis, MO, USA) or EMD Millipore, and all enzymes, unless otherwise noted, were obtained from New England Biolabs (Ipswich, MA, USA). 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP) was obtained from TCI America (Portland, OR, USA) and [γ-32P]ATP was purchased from Perkin Elmer (Piscataway, NJ, USA). All unmodified oligodeoxyribonucleotides (ODNs) were from Integrated DNA Technologies (IDT, Coralville, IA, USA). The 12-mer ODNs harboring a site-specifically incorporated O4-alkyldT were synthesized using conventional phosphoramidite chemistry, as described previously (14 (link)). The identified and purities of all the lesion-harboring ODNs were confirmed by liquid chromatography-mass spectrometry (LC-MS) and tandem MS (MS/MS) analyses prior to their insertion into double-stranded plasmids.
7
Synthesis and Characterization of Modified Oligonucleotides
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Unmodified oligodeoxyribonucleotides (ODNs)
used in this study were purchased from Integrated DNA Technologies
(Coralville, IA). [γ-32P]ATP was obtained from PerkinElmer
(Piscataway, NJ). Shrimp alkaline phosphatase (SAP) was obtained from
USB Corporation (Cleveland, OH). All other enzymes unless otherwise
specified were purchased from New England BioLabs (Ipswich, MA). 1,1,1,3,3,3-Hexafluoro-2-propanol
(HFIP) was purchased from TCI America (Portland, OR). Chemicals unless
otherwise noted were obtained from Sigma-Aldrich (St. Louis, MO).
The HEK293T human embryonic kidney epithelial cells were purchased
from ATCC (Manassas, VA). The siRNAs used in this study were purchased
from Thermo Dharmacon: human-TDG SMARTpool (L-003780)
and siRNA Control Non-Targeting pool (D-001210).
Modified cytosine-containing
ODNs (5′-ATGGCGX GCTAT-3′, X = 5mdC,
5hmdC, 5fdC, or 5cadC) were synthesized previously. The HPLC traces
for monitoring the purities of these ODNs are shown in Figure S1 (Supporting Information ), and the identities of
these ODNs were confirmed by electrospray ionization–mass spectrometry
(ESI-MS) and tandem MS (MS/MS) analyses.22 (link)
used in this study were purchased from Integrated DNA Technologies
(Coralville, IA). [γ-32P]ATP was obtained from PerkinElmer
(Piscataway, NJ). Shrimp alkaline phosphatase (SAP) was obtained from
USB Corporation (Cleveland, OH). All other enzymes unless otherwise
specified were purchased from New England BioLabs (Ipswich, MA). 1,1,1,3,3,3-Hexafluoro-2-propanol
(HFIP) was purchased from TCI America (Portland, OR). Chemicals unless
otherwise noted were obtained from Sigma-Aldrich (St. Louis, MO).
The HEK293T human embryonic kidney epithelial cells were purchased
from ATCC (Manassas, VA). The siRNAs used in this study were purchased
from Thermo Dharmacon: human-TDG SMARTpool (L-003780)
and siRNA Control Non-Targeting pool (D-001210).
Modified cytosine-containing
ODNs (5′-ATGGCG
5hmdC, 5fdC, or 5cadC) were synthesized previously. The HPLC traces
for monitoring the purities of these ODNs are shown in Figure S1 (
these ODNs were confirmed by electrospray ionization–mass spectrometry
(ESI-MS) and tandem MS (MS/MS) analyses.22 (link)
8
Synthesizing O2-alkyldT-containing Oligonucleotides
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All chemicals, unless otherwise specified, were from Sigma-Aldrich (St. Louis, MO) or EMD Millipore. 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP) was obtained from TCI America (Portland, OR). Shrimp alkaline phosphatase and [γ-32P]ATP were purchased from USB Corporation (Cleveland, OH) and Perkin Elmer (Piscataway, NJ), respectively, and all other enzymes were obtained from New England Biolabs (Ipswich, MA). All unmodified oligodeoxyribonucleotides (ODNs) were purchased from Integrated DNA Technologies (Coralville, IA). The 12-mer ODNs harboring a site-specifically incorporated O2-alkyldT were synthesized using conventional phosphoramidite chemistry, as detailed below.
M13mp7(L2) plasmid and wild-type AB1157 E. coli strains were kindly provided by Prof. John M. Essigmann, and polymerase-deficient AB1157 strains [Δpol B1::spec (Pol II-deficient), ΔdinB (Pol IV-deficient), ΔumuC::kan (Pol V-deficient), and Δpol B1::spec ΔdinB ΔumuC::kan (Pol II, Pol IV, Pol V-triple knockout)] were generously provided by Prof. Graham C. Walker.
M13mp7(L2) plasmid and wild-type AB1157 E. coli strains were kindly provided by Prof. John M. Essigmann, and polymerase-deficient AB1157 strains [Δpol B1::spec (Pol II-deficient), ΔdinB (Pol IV-deficient), ΔumuC::kan (Pol V-deficient), and Δpol B1::spec ΔdinB ΔumuC::kan (Pol II, Pol IV, Pol V-triple knockout)] were generously provided by Prof. Graham C. Walker.
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