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Phenylmethylsulfonyl fluoride pmsf

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Phenylmethylsulfonyl fluoride (PMSF) is a protease inhibitor commonly used in biochemical and molecular biology applications. It functions by irreversibly inhibiting serine proteases, which are enzymes that cleave peptide bonds in proteins. PMSF is often utilized in sample preparation processes to prevent proteolytic degradation of target proteins.

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Lab products found in correlation

Sodium chloride (nacl), by Thermo Fisher Scientific (2 mentions) 1 6 diphenyl 1 3 5 hexatriene dph, by Merck Group (1 mentions) Potassium chloride (kcl), by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 labeled annexin 5, by Thermo Fisher Scientific (1 mentions) Kh2po4, by Thermo Fisher Scientific (1 mentions) Na2hpo4, by Thermo Fisher Scientific (1 mentions) Dulbecco s pbs dpbs, by Thermo Fisher Scientific (1 mentions) 1 2 dioleoyl sn glycero 3 phosphoethanolamine n lissamine rhodamine b sulfonyl rho dope, by Avanti Polar Lipids (1 mentions) Cell culture reagents, by Euroclone (1 mentions) Yeast extract, by Thermo Fisher Scientific (1 mentions) Ampicillin, by Thermo Fisher Scientific (1 mentions) Imidazole, by Thermo Fisher Scientific (1 mentions) Isopropyl β d thiogalactopyranoside, by GoldBio (1 mentions) Sodium hydroxide (naoh), by Thermo Fisher Scientific (1 mentions) Kanamycin, by GoldBio (1 mentions) Ampicillin, by GoldBio (1 mentions) Isopropyl β d 1 thiogalactopyranoside iptg, by GoldBio (1 mentions) Hplc grade methanol, by Merck Group (1 mentions) Acetonitrile, by Merck Group (1 mentions)

4 protocols using phenylmethylsulfonyl fluoride pmsf

1

Lipid Membrane Composition Analysis

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POPC, egg SM (eSM), Cholesterol (CHOL), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (Rho-DOPE) were from Avanti Polar Lipids (Alabaster, AL). 1,6-diphenyl-1,3,5-hexatriene (DPH) was from Sigma-Aldrich (St. Louis, MO). Octydecylrhodamine B (ODRB), 3,3′-dilinoleyloxacarbocyanine perchlorate (FAST DiO), dithiothreitol (DTT), and 4 (w/v) % PFA were from Invitrogen (Carlsbad, CA). All lipids and lipid probes were stored at −20°C. NEM was from Sigma-Aldrich dissolved in methanol and stored as aliquots at −20°C. Proteinase K (ProK) was from Thermo Fisher (Bohemia, NY). Phenylmethylsulfonyl fluoride (PMSF) was from Gold Biotechnology (St. Louis, MO). Octyl glucoside (OG) was from Anatrace (Maumee, OH) and dissolved in water. 10× phosphate-buffered saline (PBS) was from Bio-RAD (Hercules, CA). Dulbecco’s PBS (DPBS) (200 mg/l KCl, 200 mg/l KH2PO4, 8 g/l NaCl, and 2.16 g/l Na2HPO4) was from Thermo Fisher Scientific (Waltham, MA). Alexa Fluor 488 labeled Annexin V was from Life Technologies (Carlsbad, CA) and 5× annexin binding buffer from Thermo Fisher Scientific. All other chemicals were reagent grade.
Kakuda S., Suresh P., Li G, & London E. (2021). Loss of plasma membrane lipid asymmetry can induce ordered domain (raft) formation. Journal of Lipid Research, 63(1), 100155.
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2

Purification of Membrane Proteins

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Cell culture reagents were purchased from EuroClone (Milan, Italy). Protoporphirin IX (Hemin), trizma base, potassium chloride, imidazole, sodium citrate tribasic dihydrate and lithium chloride, Amicon Ultra-4 10 kDa MWCO centrifugal filter device was purchased from Merck. Nickel-NTA agarose beads (low density), n-octyl β-D-glucopyranoside (β-OG) and phenylmethylsulfonyl fluoride (PMSF) were from Gold Biotechnology. Hexaethylene Glycol Monodecyl Ether (C10E6) detergent was purchased from Anatrace (Maumee, OH). Phusion High Fidelity (HF) DNA was from Biolabs. Sf-900 II SFM medium, HALT protease inhibitor single-use cocktail EDTA-free, Pierce BCA protein assay kit, Snakeskin dialysis tubing (10 kDa MWCO, 35 mm), and glycerol were purchased from Thermo Fisher Scientific Italia (Monza, Italy). TGX 10% precast polyacrylamide gels and all reagents for SDS-PAGE were purchased from BIO-RAD Laboratories Srl (Milan, Italy). Tobacco Etch Virus (TEV) protease was produced as described33 (link). Oxygraph buffer: 200 mM Tris–HCl (pH 8.3), 2 M KCl, 1 mM phenol.
Miciaccia M., Belviso B.D., Iaselli M., Cingolani G., Ferorelli S., Cappellari M., Loguercio Polosa P., Perrone M.G., Caliandro R, & Scilimati A. (2021). Three-dimensional structure of human cyclooxygenase (hCOX)-1. Scientific Reports, 11, 4312.
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3

Comprehensive Reagent Catalog for Biomolecular Assays

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Milli-Q Ultrapure water was used for all experiments. The following reagents were purchased from Millipore-Sigma: D,L-5-hydroxylysine hydrochloride, sodium metaperiodate, Dowex 50WX4 hydrogen form (200–400 mesh) cation exchange resin, sodium phosphate buffer, ε-amino-N-caproic acid, N-tosyl-L-phenylalanine chloromethyl ketone, bovine serum albumin, NADPH, NADH, and ammonium fluoride. The following reagents were from Fisher Scientific: LB Agar Miller, ampicillin, yeast extract, NaOH, NaCl, 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES), and ethylenediamine tetraacetic acid (EDTA) disodium salt. Glycerol and tryptone were purchased from Research Products International. Reagents β-octyl-D-glucopyranoside (β-OG) and L-proline were purchased from Combi-Blocks. Ethanol (190- or 200-proof) was purchased from Decon Labs. Tris(hydroxymethyl)aminomethane (Tris), glycine, isopropyl β-D-thiogalactopyranoside (IPTG), phenylmethylsulfonyl fluoride (PMSF), leupeptin hemisulfate, and NAD+ were purchased from Gold Biotechnology. Imidazole and (O)2-aminobenzaldehyde (o-AB) were purchased from Acros Organics. Hydrochloric acid (HCl) 34–37% OmniTrace was purchased from EMD-Millipore.
Patel S.M., Seravalli J., Liang X., Tanner J.J, & Becker D.F. (2021). Disease Variants of Human Δ1-Pyrroline-5-Carboxylate Reductase 2 (PYCR2). Archives of biochemistry and biophysics, 703, 108852.
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4

Reagent Sources for Amino Acid Synthesis

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The N-α-Fmoc-protected amino acids were purchased from either Novabiochem or ChemPep Inc. HPLC grade methanol and acetonitrile were purchased from either Sigma Aldrich or British Drug Houses (BDH). Phenylmethylsulfonyl fluoride (PMSF) was purchased from either Gold Biotechnology or Sigma Aldrich. Kanamycin, ampicillin, and isopropyl β-D-1-thiogalactopyranoside (IPTG) were purchased from Gold Biotechnology. Unless otherwise stated, the remaining chemical reagents described were purchased from Fisher Scientific, Acros Organics, Sigma Aldrich, Alfa Aesar, BDH, Research Products International Corp., Macron Fine Chemicals, Bio-Rad, or J.T. Baker.
Fulton M.D., Zhang J., He M., Ho M.C, & Zheng Y.G. (2017). The Intricate Effects of alpha-Amino and Lysine Modifications on Arginine Methylation on the N-terminal Tail of Histone H4. Biochemistry, 56(28), 3539-3548.
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