iodoacetamide  (Millipore)


Bioz Verified Symbol Millipore is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99

    Structured Review

    Millipore iodoacetamide
    Iodoacetamide, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 294 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/iodoacetamide/product/Millipore
    Average 99 stars, based on 294 article reviews
    Price from $9.99 to $1999.99
    iodoacetamide - by Bioz Stars, 2020-02
    99/100 stars

    Images

    Related Articles

    Bradford Assay:

    Article Title: STIM2 drives Ca2+ oscillations through store-operated Ca2+ entry caused by mild store depletion
    Article Snippet: RBL-2H3 and HEK293 cells were harvested at the indicated time points after transfection, washed in PBS and lysed in 1 ml lysis buffer (in m m ): 75 NaCl, 40 NaF, 10 Iodocetamide, 50 Hepes, 10% IGEPAL, 1 phenylmethylsulphonyl fluoride and protease inhibitor cocktail (Sigma). .. The protein concentrations from HEK293 cells lysates (see Supplementary ) were determined using the Bradford assay.

    Stripping Membranes:

    Article Title: A comparative method for protein extraction and 2-D gel electrophoresis from different tissues of Cajanus cajan
    Article Snippet: .. 2-Mercaptoethanol (2-ME, Sigma-Aldrich, cat. no. M6250), 3-[(3-Cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS, Sigma-Aldrich, cat. no. C3023), Acetone (Sigma-Aldrich, cat. no. 650501), Acetic acid (Sigma-Aldrich, cat. no. V800018), Agarose (Sigma-Aldrich, cat. no. A9539), Ammonium acetate (Sigma-Aldrich, cat. no. A7262), Bradford reagent (Sigma-Aldrich, cat. no. B6916), Bromophenol blue (BPB, Sigma-Aldrich, cat. no. B0126), Bovine serum albumin (BSA, Sigma-Aldrich, cat. no. 05470), Coomassie brilliant blue (CBB, Bioworld, cat. no. 742083), Dithiothreitol (DTT, Sigma-Aldrich, cat. no. D9779), Ethylene diamine tetra-acetic acid, 0.5 M, pH 8.0 (EDTA, Sigma-Aldrich, cat. no. E7889), Glycerol (Sigma-Aldrich, cat. no. G5516), Glycine (Sigma-Aldrich, cat. no. 50046), Hydrogen chloride (HCl, Sigma-Aldrich, cat. no. H1758), ImmobilineTM Dry strip, pH 3-10, 13 cm (GE Healthcare, UK, cat. no. 17-6001-14), IPG buffer, pH 3-10 NL (GE Healthcare, UK, cat. no. 17-6000-88), Iodoacetamide (IAA, Sigma-Aldrich, cat. no. I3750), Methanol (Sigma-Aldrich, cat. no. 322415), Phenyl methane sulfonyl fluoride (PMSF, Sigma-Aldrich, cat. no. P7626), Potassium chloride (KCl, Sigma-Aldrich, cat. no. P9541), Phosphate buffer, 1M, pH 7.5 (Sigma-Aldrich, cat. no P3619), Complete EDTA free Protease inhibitor cocktail (Roche, cat. no. REF 11 875 580 001), Sodium dodecyl sulfate (SDS, Sigma-Aldrich, cat. no. L4390), Sucrose (Sigma-Aldrich, cat. no. S9378), Thiourea (Sigma-Aldrich, cat. no. T8656), Trichloroacetic acid (TCA; Sigma-Aldrich, cat. no. 522082), Tris (Trizma base, Sigma-Aldrich, cat. no. T1503), Tris-buffered phenol solution (stored at 4°C, pH 7.8–8.0; Sigma-Aldrich, cat. no. P 4557), Tris-HCl, 1.5 M, pH 8.8 (Amresco, cat. no. M195), Urea (Sigma-Aldrich, cat. no. U6504). ..

    Mass Spectrometry:

    Article Title: An Effective Protocol for Proteome Analysis of Medaka (Oryzias latipes) after Acute Exposure to Ionizing Radiation
    Article Snippet: .. Materials 12 wild-type adult Medaka (~6–8 months old) with a body weight of 0.45 + 0.01 g and a 14 h:10 h light-dark cycle at 25 °C; 4 liter of filtered water; Anesthetic solution: 250 mg/L of tricaine methane sulfonate (MS-222) (Fisher, Fair Lawn, NJ, USA; cat: NC0342409), buffered with sodium bicarbonate at or near to neutral pH; Liquid nitrogen; Methanol HPLC grade (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: 34860-4L-R); Homogenizing solution kept at 4 °C: Mix chloroform (Fisher, Fair Lawn, NJ, USA; Cat. no.: C606SK-4), methanol, and Milli-Q water at ratio of 2:4:1.5, respectively; Acetone (Fisher, Fair Lawn, NJ, USA; Cat. no.: C606SK-4); Tris-HCl buffer: 1 mM Tris base (Fisher Scientific, Fair Lawn, NJ, USA; Cat. no.: BP152-500), pH 7.2 and 2% SDS (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: L4390); Pierce BCA protein assay kit (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: 23227); Bolt 4–12% Bis-Tris Plus gels (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: NW04120BOX); Protein ladder (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: LC5925); 2× Laemmli Sample Buffer (Bio-Rad, Hercules, CA, USA; Cat. no.: 1610737); Running buffer 1×: 50 mL of Bolt MOPS SDS running buffer (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: B0001) mixed with 950 mL of deionized H2 O (dH2O); Plastic container 12 × 8 × 3 cm, rinsed previously with 50% aqueous methanol twice and one time with 100% methanol; Instant Blue Coomassie (Expedeon, Cambridge, UK; Cat. no: ISB1L); Destaining solution: 50 mL methanol, 5 mL acetic acid (J.T Baker, Center Valley, PA, USA; Cat. no.: 9508-33) and 50 mL dH2O; Isopropanol (Fisher, Fair Lawn, NJ, USA; Cat. no.: BP2635-4); Ambic solution: 0.158 g of ammonium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: AG141-500G) dissolved in 20 mL of dH2O to obtain a 100 mM solution; Acetonitrile HPLC grade (ACN) (Fisher, Fair Lawn, NJ, USA; Cat. no.: A988-4); 10 mM dithiothreitol (DTT): 0.015 g of DTT (Research Products International, Mount Prospect, IL, USA; Cat. no.: D11000-25) dissolved in 10 mL of Ambic solution; 55 mM iodoacetamide (IDA): 0.10 g of iodoacetamide (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: I1149-5G) dissolved in 10 mL of Ambic solution; Trypsin solution: 20 µg of trypsin (Promega, Madison, WI, USA; Cat. no.: V5111) in 1 mL of cold Ambic solution. .. Final concentration 20 ng/µL; Extraction solution: 50% ACN / 50% dH2O and 0.1% formic acid (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: F0507-500mL); Buffer A (0.1% aqueous formic acid); Buffer B (80% ACN, 0.1% aqueous formic acid).

    Article Title: Prmt5 is a regulator of muscle stem cell expansion in adult mice
    Article Snippet: Paragraph title: Mass spectrometry ... Gel pieces were reduced with 10 mM dithiothreitol for 45 min at 56 °C and alkylated with 55 mM iodocetamide (BioUltra, Sigma-Aldrich) for 30 min at room temperature in the dark.

    BIA-KA:

    Article Title: An Effective Protocol for Proteome Analysis of Medaka (Oryzias latipes) after Acute Exposure to Ionizing Radiation
    Article Snippet: .. Materials 12 wild-type adult Medaka (~6–8 months old) with a body weight of 0.45 + 0.01 g and a 14 h:10 h light-dark cycle at 25 °C; 4 liter of filtered water; Anesthetic solution: 250 mg/L of tricaine methane sulfonate (MS-222) (Fisher, Fair Lawn, NJ, USA; cat: NC0342409), buffered with sodium bicarbonate at or near to neutral pH; Liquid nitrogen; Methanol HPLC grade (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: 34860-4L-R); Homogenizing solution kept at 4 °C: Mix chloroform (Fisher, Fair Lawn, NJ, USA; Cat. no.: C606SK-4), methanol, and Milli-Q water at ratio of 2:4:1.5, respectively; Acetone (Fisher, Fair Lawn, NJ, USA; Cat. no.: C606SK-4); Tris-HCl buffer: 1 mM Tris base (Fisher Scientific, Fair Lawn, NJ, USA; Cat. no.: BP152-500), pH 7.2 and 2% SDS (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: L4390); Pierce BCA protein assay kit (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: 23227); Bolt 4–12% Bis-Tris Plus gels (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: NW04120BOX); Protein ladder (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: LC5925); 2× Laemmli Sample Buffer (Bio-Rad, Hercules, CA, USA; Cat. no.: 1610737); Running buffer 1×: 50 mL of Bolt MOPS SDS running buffer (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: B0001) mixed with 950 mL of deionized H2 O (dH2O); Plastic container 12 × 8 × 3 cm, rinsed previously with 50% aqueous methanol twice and one time with 100% methanol; Instant Blue Coomassie (Expedeon, Cambridge, UK; Cat. no: ISB1L); Destaining solution: 50 mL methanol, 5 mL acetic acid (J.T Baker, Center Valley, PA, USA; Cat. no.: 9508-33) and 50 mL dH2O; Isopropanol (Fisher, Fair Lawn, NJ, USA; Cat. no.: BP2635-4); Ambic solution: 0.158 g of ammonium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: AG141-500G) dissolved in 20 mL of dH2O to obtain a 100 mM solution; Acetonitrile HPLC grade (ACN) (Fisher, Fair Lawn, NJ, USA; Cat. no.: A988-4); 10 mM dithiothreitol (DTT): 0.015 g of DTT (Research Products International, Mount Prospect, IL, USA; Cat. no.: D11000-25) dissolved in 10 mL of Ambic solution; 55 mM iodoacetamide (IDA): 0.10 g of iodoacetamide (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: I1149-5G) dissolved in 10 mL of Ambic solution; Trypsin solution: 20 µg of trypsin (Promega, Madison, WI, USA; Cat. no.: V5111) in 1 mL of cold Ambic solution. .. Final concentration 20 ng/µL; Extraction solution: 50% ACN / 50% dH2O and 0.1% formic acid (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: F0507-500mL); Buffer A (0.1% aqueous formic acid); Buffer B (80% ACN, 0.1% aqueous formic acid).

    Modification:

    Article Title: Prmt5 is a regulator of muscle stem cell expansion in adult mice
    Article Snippet: Gel pieces were reduced with 10 mM dithiothreitol for 45 min at 56 °C and alkylated with 55 mM iodocetamide (BioUltra, Sigma-Aldrich) for 30 min at room temperature in the dark. .. Gel pieces were digested overnight at 37 °C in 50 μl of digestion buffer containing 12.5 ng μl of Sequencing Grade Modified Trypsin (Promega Corp., Madison, USA).

    Western Blot:

    Article Title: STIM2 drives Ca2+ oscillations through store-operated Ca2+ entry caused by mild store depletion
    Article Snippet: Paragraph title: Preparation of cell extracts, immunoprecipitation and Western blotting ... RBL-2H3 and HEK293 cells were harvested at the indicated time points after transfection, washed in PBS and lysed in 1 ml lysis buffer (in m m ): 75 NaCl, 40 NaF, 10 Iodocetamide, 50 Hepes, 10% IGEPAL, 1 phenylmethylsulphonyl fluoride and protease inhibitor cocktail (Sigma).

    High Performance Liquid Chromatography:

    Article Title: An Effective Protocol for Proteome Analysis of Medaka (Oryzias latipes) after Acute Exposure to Ionizing Radiation
    Article Snippet: .. Materials 12 wild-type adult Medaka (~6–8 months old) with a body weight of 0.45 + 0.01 g and a 14 h:10 h light-dark cycle at 25 °C; 4 liter of filtered water; Anesthetic solution: 250 mg/L of tricaine methane sulfonate (MS-222) (Fisher, Fair Lawn, NJ, USA; cat: NC0342409), buffered with sodium bicarbonate at or near to neutral pH; Liquid nitrogen; Methanol HPLC grade (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: 34860-4L-R); Homogenizing solution kept at 4 °C: Mix chloroform (Fisher, Fair Lawn, NJ, USA; Cat. no.: C606SK-4), methanol, and Milli-Q water at ratio of 2:4:1.5, respectively; Acetone (Fisher, Fair Lawn, NJ, USA; Cat. no.: C606SK-4); Tris-HCl buffer: 1 mM Tris base (Fisher Scientific, Fair Lawn, NJ, USA; Cat. no.: BP152-500), pH 7.2 and 2% SDS (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: L4390); Pierce BCA protein assay kit (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: 23227); Bolt 4–12% Bis-Tris Plus gels (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: NW04120BOX); Protein ladder (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: LC5925); 2× Laemmli Sample Buffer (Bio-Rad, Hercules, CA, USA; Cat. no.: 1610737); Running buffer 1×: 50 mL of Bolt MOPS SDS running buffer (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: B0001) mixed with 950 mL of deionized H2 O (dH2O); Plastic container 12 × 8 × 3 cm, rinsed previously with 50% aqueous methanol twice and one time with 100% methanol; Instant Blue Coomassie (Expedeon, Cambridge, UK; Cat. no: ISB1L); Destaining solution: 50 mL methanol, 5 mL acetic acid (J.T Baker, Center Valley, PA, USA; Cat. no.: 9508-33) and 50 mL dH2O; Isopropanol (Fisher, Fair Lawn, NJ, USA; Cat. no.: BP2635-4); Ambic solution: 0.158 g of ammonium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: AG141-500G) dissolved in 20 mL of dH2O to obtain a 100 mM solution; Acetonitrile HPLC grade (ACN) (Fisher, Fair Lawn, NJ, USA; Cat. no.: A988-4); 10 mM dithiothreitol (DTT): 0.015 g of DTT (Research Products International, Mount Prospect, IL, USA; Cat. no.: D11000-25) dissolved in 10 mL of Ambic solution; 55 mM iodoacetamide (IDA): 0.10 g of iodoacetamide (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: I1149-5G) dissolved in 10 mL of Ambic solution; Trypsin solution: 20 µg of trypsin (Promega, Madison, WI, USA; Cat. no.: V5111) in 1 mL of cold Ambic solution. .. Final concentration 20 ng/µL; Extraction solution: 50% ACN / 50% dH2O and 0.1% formic acid (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: F0507-500mL); Buffer A (0.1% aqueous formic acid); Buffer B (80% ACN, 0.1% aqueous formic acid).

    Article Title: A comparative method for protein extraction and 2-D gel electrophoresis from different tissues of Cajanus cajan
    Article Snippet: Reagents The chemicals and the reagents used in the experiments were HPLC grade and procured from Sigma-Aldrich (≥99.93%). .. 2-Mercaptoethanol (2-ME, Sigma-Aldrich, cat. no. M6250), 3-[(3-Cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS, Sigma-Aldrich, cat. no. C3023), Acetone (Sigma-Aldrich, cat. no. 650501), Acetic acid (Sigma-Aldrich, cat. no. V800018), Agarose (Sigma-Aldrich, cat. no. A9539), Ammonium acetate (Sigma-Aldrich, cat. no. A7262), Bradford reagent (Sigma-Aldrich, cat. no. B6916), Bromophenol blue (BPB, Sigma-Aldrich, cat. no. B0126), Bovine serum albumin (BSA, Sigma-Aldrich, cat. no. 05470), Coomassie brilliant blue (CBB, Bioworld, cat. no. 742083), Dithiothreitol (DTT, Sigma-Aldrich, cat. no. D9779), Ethylene diamine tetra-acetic acid, 0.5 M, pH 8.0 (EDTA, Sigma-Aldrich, cat. no. E7889), Glycerol (Sigma-Aldrich, cat. no. G5516), Glycine (Sigma-Aldrich, cat. no. 50046), Hydrogen chloride (HCl, Sigma-Aldrich, cat. no. H1758), ImmobilineTM Dry strip, pH 3-10, 13 cm (GE Healthcare, UK, cat. no. 17-6001-14), IPG buffer, pH 3-10 NL (GE Healthcare, UK, cat. no. 17-6000-88), Iodoacetamide (IAA, Sigma-Aldrich, cat. no. I3750), Methanol (Sigma-Aldrich, cat. no. 322415), Phenyl methane sulfonyl fluoride (PMSF, Sigma-Aldrich, cat. no. P7626), Potassium chloride (KCl, Sigma-Aldrich, cat. no. P9541), Phosphate buffer, 1M, pH 7.5 (Sigma-Aldrich, cat. no P3619), Complete EDTA free Protease inhibitor cocktail (Roche, cat. no. REF 11 875 580 001), Sodium dodecyl sulfate (SDS, Sigma-Aldrich, cat. no. L4390), Sucrose (Sigma-Aldrich, cat. no. S9378), Thiourea (Sigma-Aldrich, cat. no. T8656), Trichloroacetic acid (TCA; Sigma-Aldrich, cat. no. 522082), Tris (Trizma base, Sigma-Aldrich, cat. no. T1503), Tris-buffered phenol solution (stored at 4°C, pH 7.8–8.0; Sigma-Aldrich, cat. no. P 4557), Tris-HCl, 1.5 M, pH 8.8 (Amresco, cat. no. M195), Urea (Sigma-Aldrich, cat. no. U6504).

    Transfection:

    Article Title: STIM2 drives Ca2+ oscillations through store-operated Ca2+ entry caused by mild store depletion
    Article Snippet: .. RBL-2H3 and HEK293 cells were harvested at the indicated time points after transfection, washed in PBS and lysed in 1 ml lysis buffer (in m m ): 75 NaCl, 40 NaF, 10 Iodocetamide, 50 Hepes, 10% IGEPAL, 1 phenylmethylsulphonyl fluoride and protease inhibitor cocktail (Sigma). .. The cell lysates from RBL-2H3 (see ) were precipitated with anti-STIM2(CT) rabbit polyclonal antibody (2.5 μg; Anaspec) for 1.5 h at 4°C and resolved by SDS-PAGE.

    Chromatography:

    Article Title: Prmt5 is a regulator of muscle stem cell expansion in adult mice
    Article Snippet: Gel pieces were reduced with 10 mM dithiothreitol for 45 min at 56 °C and alkylated with 55 mM iodocetamide (BioUltra, Sigma-Aldrich) for 30 min at room temperature in the dark. .. Released peptides were extracted (collecting separately the liquid mixture of each samples at each step) once by adding 100 μl of 30% acetonitrile liquid chromatography/mass spectrometry (LC/MS) grade (Thermo Scientific)/3% trifluoroacetic acid (Sigma-Aldrich) in water, and twice by adding 70% acetonitrile, followed by two final extractions with 100% acetonitrile.

    Immunoprecipitation:

    Article Title: STIM2 drives Ca2+ oscillations through store-operated Ca2+ entry caused by mild store depletion
    Article Snippet: Paragraph title: Preparation of cell extracts, immunoprecipitation and Western blotting ... RBL-2H3 and HEK293 cells were harvested at the indicated time points after transfection, washed in PBS and lysed in 1 ml lysis buffer (in m m ): 75 NaCl, 40 NaF, 10 Iodocetamide, 50 Hepes, 10% IGEPAL, 1 phenylmethylsulphonyl fluoride and protease inhibitor cocktail (Sigma).

    Protease Inhibitor:

    Article Title: STIM2 drives Ca2+ oscillations through store-operated Ca2+ entry caused by mild store depletion
    Article Snippet: .. RBL-2H3 and HEK293 cells were harvested at the indicated time points after transfection, washed in PBS and lysed in 1 ml lysis buffer (in m m ): 75 NaCl, 40 NaF, 10 Iodocetamide, 50 Hepes, 10% IGEPAL, 1 phenylmethylsulphonyl fluoride and protease inhibitor cocktail (Sigma). .. The cell lysates from RBL-2H3 (see ) were precipitated with anti-STIM2(CT) rabbit polyclonal antibody (2.5 μg; Anaspec) for 1.5 h at 4°C and resolved by SDS-PAGE.

    Article Title: Site-specific chemical protein conjugation using genetically encoded aldehyde tags
    Article Snippet: .. Acetic acid (AcOH; Fisher Scientific, cat. no. A38-212) Acetic acid in deionized water (DI H2 O; 7.5%, vol/vol) Acetonitrile (ACN; Sigma-Aldrich, cat. no. 271004) Agar (Fisher Scientific, cat. no. BP2641) Alexa Fluor 647 C5-aminooxyacetamide (Molecular Probes, cat. no. ) Alexa Fluor 647 C5-aminooxyacetamide (10 mM in DMSO) Alkylation buffer Annealing buffer l -Arabinose (Sigma-Aldrich Chemical, cat. no. A3256) l -arabinose (20%, wt/vol) BamHI (NEB, cat. no. R0136) BglII (NEB, cat. no. R0104) Biomax 5K filter (Millipore, cat. no. UFV5BCC00) Bromophenol blue (Sigma-Aldrich Chemical, cat. no. B8026) Calf intestinal alkaline phosphatase (CIP; NEB, cat. no. M0290) Carbenicillin (Sigma-Aldrich Chemical, cat. no. C9231) CHO K1 cells (ATCC, cat. no. CCL-61) Complete EDTA-free protease inhibitor cocktail (Roche, cat. no. 11 873 580 001) Criterion XT MES running buffer (20×; Bio-Rad, cat. no. 161-0789) 1× Criterion XT MES running buffer Deionized water (DI H2 O) Denaturing buffer Dimethyl sulfoxide (DMSO; Sigma-Aldrich Chemical, cat. no. D8418) Dithiothreitol (DTT; Sigma-Aldrich Chemical, cat. no. 43815) DnaseI (NEB, cat. no. M0303) DpnI (NEB, cat. no. R0176) dNTPs (2.5 mM; Invitrogen, cat. no. ) EcoRV (NEB, cat. no. R0104) Elution buffer Ethylenediamine tetraacidic disodium salt (EDTA; Fisher Scientific, cat. no. BP120-1) Ex-Cell 325 PF CHO serum-free medium (Sigma-Aldrich, cat. no. 14340C) Fetal bovine serum (Mediatech, cat. no. 35-010-CV) l -Glutamine (Mediatech, cat. no. 25-005-CI) Glycerol (Sigma-Aldrich Chemical, cat. no. G5516) Glycerol (50%, vol/vol) Guanidine hydrochloride (Sigma-Aldrich, cat. no. G4505) HindIII (NEB, cat. no. R0104) Hydrochloric acid (HCl; EMD, cat. no. 3120-500) IgG elution buffer (Thermo Fisher Scientific, cat. no. 21004) Imidazole (Sigma-Aldrich Chemical, cat. no. I2399) Iodoacetamide (IAA; Sigma-Aldrich, cat. no. 068k53022) Isopropyl β- d -1-thiogalactopyranoside (IPTG; Sigma-Aldrich Chemical, cat. no. I5502) IPTG (isopropyl-1-thio-β- d -galactopyranoside, 1 M) Kanamycin (Sigma-Aldrich Chemical, cat. no. K1876) Labeling buffer (2×, 500 mM NaOAc, pH = 4.6) Lipofectamine 2000 (Invitrogen, cat. no. 11668019) Liquid nitrogen Lysis buffer Lysozyme (Sigma-Aldrich Chemical, cat. no. L6876) Methoxyamine hydrochloride (Sigma-Aldrich, cat. no. M1139) Ni-NTA agarose (Qiagen, cat. no. 30230) Oligonucleotides (see ) .. Amicon Ultra centrifugal filters (10 kDa molecular weight cutoff, Millipore, cat. no. UFC5010BK) Benchtop centrifuge (Sorvall Legend Mach 1.6 refrigerated, cat. no. 75004337) Centrifuge (Sorvall RC5C plus, cat. no. 74505) Criterion cell (Bio-Rad, cat. no. 165-6001) Criterion XT 4–12% (wt/vol) Bis-Tris gel (Bio-Rad, cat. no. 345-0123) Falcon tubes (15 ml; Fisher Scientific, cat. no. 05-527-90) Falcon tubes (50 ml, Fisher Scientific, cat. no. 14-432-22) Fast protein liquid chromatography (FPLC; GE Healthcare, ÄKTA purifier) HPLC (Waters) Microcentrifuge (Eppendorf 5417c, cat. no. 5417 000.315) Microcentrifuge tubes (2 ml; Fisher Scientific, cat. no. 02-681-291) NanoVue spectrophotometer (GE) PCR machine (Eppendorf, Mastercycler gradient, cat. no. 950000015) PCR tubes (Fisher Scientific, cat. no. 14-230-225) PD SpinTrap G-25 (GE Healthcare Life Sciences, cat. no. 28-9180-04) Reverse-phase HPLC column (Phenomenex, part: 00G-4396-B0) Shaking incubator (set at 220 r.p.m., refrigerated 4230; New Brunswick Scientific, cat. no. M1335-0012) Sterile 16-ml culture tubes (Fisher Scientific, cat. no. 05-540-6) Sterile 100 mm × 25 mm Petri dishes (Fisher Scientific, cat. no. 08-757-13) Sterile 0.22-μm polyethersulfone filters (Millipore, cat. no. SLGP033RS) Spin concentrator (Amicon Ultra 30kDa; Millipore, cat. no. UFC903008) Sonicator (Misonix sonicator 3000; Misonix) Tabletop rocking platform (Fisher Scientific, cat. no. 1404123Q) Typhoon variable mode imager (Amersham Bioscience, cat. no. 63-0055-80) Vortex mixer (Vortex Genie-2, Mo Bio Laboratories, cat. no. 13111) Waters QToF premier mass spectrometer (Waters) and Masslynx software

    Article Title: A comparative method for protein extraction and 2-D gel electrophoresis from different tissues of Cajanus cajan
    Article Snippet: .. 2-Mercaptoethanol (2-ME, Sigma-Aldrich, cat. no. M6250), 3-[(3-Cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS, Sigma-Aldrich, cat. no. C3023), Acetone (Sigma-Aldrich, cat. no. 650501), Acetic acid (Sigma-Aldrich, cat. no. V800018), Agarose (Sigma-Aldrich, cat. no. A9539), Ammonium acetate (Sigma-Aldrich, cat. no. A7262), Bradford reagent (Sigma-Aldrich, cat. no. B6916), Bromophenol blue (BPB, Sigma-Aldrich, cat. no. B0126), Bovine serum albumin (BSA, Sigma-Aldrich, cat. no. 05470), Coomassie brilliant blue (CBB, Bioworld, cat. no. 742083), Dithiothreitol (DTT, Sigma-Aldrich, cat. no. D9779), Ethylene diamine tetra-acetic acid, 0.5 M, pH 8.0 (EDTA, Sigma-Aldrich, cat. no. E7889), Glycerol (Sigma-Aldrich, cat. no. G5516), Glycine (Sigma-Aldrich, cat. no. 50046), Hydrogen chloride (HCl, Sigma-Aldrich, cat. no. H1758), ImmobilineTM Dry strip, pH 3-10, 13 cm (GE Healthcare, UK, cat. no. 17-6001-14), IPG buffer, pH 3-10 NL (GE Healthcare, UK, cat. no. 17-6000-88), Iodoacetamide (IAA, Sigma-Aldrich, cat. no. I3750), Methanol (Sigma-Aldrich, cat. no. 322415), Phenyl methane sulfonyl fluoride (PMSF, Sigma-Aldrich, cat. no. P7626), Potassium chloride (KCl, Sigma-Aldrich, cat. no. P9541), Phosphate buffer, 1M, pH 7.5 (Sigma-Aldrich, cat. no P3619), Complete EDTA free Protease inhibitor cocktail (Roche, cat. no. REF 11 875 580 001), Sodium dodecyl sulfate (SDS, Sigma-Aldrich, cat. no. L4390), Sucrose (Sigma-Aldrich, cat. no. S9378), Thiourea (Sigma-Aldrich, cat. no. T8656), Trichloroacetic acid (TCA; Sigma-Aldrich, cat. no. 522082), Tris (Trizma base, Sigma-Aldrich, cat. no. T1503), Tris-buffered phenol solution (stored at 4°C, pH 7.8–8.0; Sigma-Aldrich, cat. no. P 4557), Tris-HCl, 1.5 M, pH 8.8 (Amresco, cat. no. M195), Urea (Sigma-Aldrich, cat. no. U6504). ..

    Cell Culture:

    Article Title: Identification of a novel anoikis signalling pathway using the fungal virulence factor gliotoxin
    Article Snippet: C3 toxin is a binary toxin consisting of C2II (200 ng/ml) and C2I/C3 (100 ng/ml) mixed in cell culture media. .. The FAK inhibitor 14 (FAK14) was purchased from Tocris (Bristol, UK), and Cilengitide, a cyclic pentapeptide RGD compound (cyclo-[RGDfN(Me)V]) , the RGD peptide, DTT and iodocetamide were from Sigma (Taufkirchen, Germany).

    Sequencing:

    Article Title: Prmt5 is a regulator of muscle stem cell expansion in adult mice
    Article Snippet: Gel pieces were reduced with 10 mM dithiothreitol for 45 min at 56 °C and alkylated with 55 mM iodocetamide (BioUltra, Sigma-Aldrich) for 30 min at room temperature in the dark. .. Gel pieces were digested overnight at 37 °C in 50 μl of digestion buffer containing 12.5 ng μl of Sequencing Grade Modified Trypsin (Promega Corp., Madison, USA).

    Nucleic Acid Electrophoresis:

    Article Title: Proteomic Analysis of Kidney Preservation Solutions Prior to Renal Transplantation
    Article Snippet: Materials The 2D electrophoresis equipment, i.e., isoelectric focusing (PROTEAN IEF) and Sodium dodecyl sulfate—polyacrylamide gel electrophoresis (protean II XI 2-D cell), the immobilized pH gradient (IPG; BioLyte) strips, mineral oil, glycine, ampholytes, and Ready Prep 2D clean-up kit were obtained from Bio-Rad (Hercules, CA, USA). .. SDS, tris, urea, thiourea, ammonium bicarbonate, 3-[(3-cholamido propyl) dimethylamonio]-1-propanesulfonate (CHAPS), isopropanol, iodoacetamide, acrylamide, bisacrylamide, the ProteoSilver Plus Silver Stain Kit, glycerol, and α-cyanohydroxycinnamic acid were obtained from Sigma-Aldrich (St. Louis, MO, USA).

    Two-Dimensional Gel Electrophoresis:

    Article Title: Proteomic Analysis of Kidney Preservation Solutions Prior to Renal Transplantation
    Article Snippet: Materials The 2D electrophoresis equipment, i.e., isoelectric focusing (PROTEAN IEF) and Sodium dodecyl sulfate—polyacrylamide gel electrophoresis (protean II XI 2-D cell), the immobilized pH gradient (IPG; BioLyte) strips, mineral oil, glycine, ampholytes, and Ready Prep 2D clean-up kit were obtained from Bio-Rad (Hercules, CA, USA). .. SDS, tris, urea, thiourea, ammonium bicarbonate, 3-[(3-cholamido propyl) dimethylamonio]-1-propanesulfonate (CHAPS), isopropanol, iodoacetamide, acrylamide, bisacrylamide, the ProteoSilver Plus Silver Stain Kit, glycerol, and α-cyanohydroxycinnamic acid were obtained from Sigma-Aldrich (St. Louis, MO, USA).

    Labeling:

    Article Title: Site-specific chemical protein conjugation using genetically encoded aldehyde tags
    Article Snippet: .. Acetic acid (AcOH; Fisher Scientific, cat. no. A38-212) Acetic acid in deionized water (DI H2 O; 7.5%, vol/vol) Acetonitrile (ACN; Sigma-Aldrich, cat. no. 271004) Agar (Fisher Scientific, cat. no. BP2641) Alexa Fluor 647 C5-aminooxyacetamide (Molecular Probes, cat. no. ) Alexa Fluor 647 C5-aminooxyacetamide (10 mM in DMSO) Alkylation buffer Annealing buffer l -Arabinose (Sigma-Aldrich Chemical, cat. no. A3256) l -arabinose (20%, wt/vol) BamHI (NEB, cat. no. R0136) BglII (NEB, cat. no. R0104) Biomax 5K filter (Millipore, cat. no. UFV5BCC00) Bromophenol blue (Sigma-Aldrich Chemical, cat. no. B8026) Calf intestinal alkaline phosphatase (CIP; NEB, cat. no. M0290) Carbenicillin (Sigma-Aldrich Chemical, cat. no. C9231) CHO K1 cells (ATCC, cat. no. CCL-61) Complete EDTA-free protease inhibitor cocktail (Roche, cat. no. 11 873 580 001) Criterion XT MES running buffer (20×; Bio-Rad, cat. no. 161-0789) 1× Criterion XT MES running buffer Deionized water (DI H2 O) Denaturing buffer Dimethyl sulfoxide (DMSO; Sigma-Aldrich Chemical, cat. no. D8418) Dithiothreitol (DTT; Sigma-Aldrich Chemical, cat. no. 43815) DnaseI (NEB, cat. no. M0303) DpnI (NEB, cat. no. R0176) dNTPs (2.5 mM; Invitrogen, cat. no. ) EcoRV (NEB, cat. no. R0104) Elution buffer Ethylenediamine tetraacidic disodium salt (EDTA; Fisher Scientific, cat. no. BP120-1) Ex-Cell 325 PF CHO serum-free medium (Sigma-Aldrich, cat. no. 14340C) Fetal bovine serum (Mediatech, cat. no. 35-010-CV) l -Glutamine (Mediatech, cat. no. 25-005-CI) Glycerol (Sigma-Aldrich Chemical, cat. no. G5516) Glycerol (50%, vol/vol) Guanidine hydrochloride (Sigma-Aldrich, cat. no. G4505) HindIII (NEB, cat. no. R0104) Hydrochloric acid (HCl; EMD, cat. no. 3120-500) IgG elution buffer (Thermo Fisher Scientific, cat. no. 21004) Imidazole (Sigma-Aldrich Chemical, cat. no. I2399) Iodoacetamide (IAA; Sigma-Aldrich, cat. no. 068k53022) Isopropyl β- d -1-thiogalactopyranoside (IPTG; Sigma-Aldrich Chemical, cat. no. I5502) IPTG (isopropyl-1-thio-β- d -galactopyranoside, 1 M) Kanamycin (Sigma-Aldrich Chemical, cat. no. K1876) Labeling buffer (2×, 500 mM NaOAc, pH = 4.6) Lipofectamine 2000 (Invitrogen, cat. no. 11668019) Liquid nitrogen Lysis buffer Lysozyme (Sigma-Aldrich Chemical, cat. no. L6876) Methoxyamine hydrochloride (Sigma-Aldrich, cat. no. M1139) Ni-NTA agarose (Qiagen, cat. no. 30230) Oligonucleotides (see ) .. Amicon Ultra centrifugal filters (10 kDa molecular weight cutoff, Millipore, cat. no. UFC5010BK) Benchtop centrifuge (Sorvall Legend Mach 1.6 refrigerated, cat. no. 75004337) Centrifuge (Sorvall RC5C plus, cat. no. 74505) Criterion cell (Bio-Rad, cat. no. 165-6001) Criterion XT 4–12% (wt/vol) Bis-Tris gel (Bio-Rad, cat. no. 345-0123) Falcon tubes (15 ml; Fisher Scientific, cat. no. 05-527-90) Falcon tubes (50 ml, Fisher Scientific, cat. no. 14-432-22) Fast protein liquid chromatography (FPLC; GE Healthcare, ÄKTA purifier) HPLC (Waters) Microcentrifuge (Eppendorf 5417c, cat. no. 5417 000.315) Microcentrifuge tubes (2 ml; Fisher Scientific, cat. no. 02-681-291) NanoVue spectrophotometer (GE) PCR machine (Eppendorf, Mastercycler gradient, cat. no. 950000015) PCR tubes (Fisher Scientific, cat. no. 14-230-225) PD SpinTrap G-25 (GE Healthcare Life Sciences, cat. no. 28-9180-04) Reverse-phase HPLC column (Phenomenex, part: 00G-4396-B0) Shaking incubator (set at 220 r.p.m., refrigerated 4230; New Brunswick Scientific, cat. no. M1335-0012) Sterile 16-ml culture tubes (Fisher Scientific, cat. no. 05-540-6) Sterile 100 mm × 25 mm Petri dishes (Fisher Scientific, cat. no. 08-757-13) Sterile 0.22-μm polyethersulfone filters (Millipore, cat. no. SLGP033RS) Spin concentrator (Amicon Ultra 30kDa; Millipore, cat. no. UFC903008) Sonicator (Misonix sonicator 3000; Misonix) Tabletop rocking platform (Fisher Scientific, cat. no. 1404123Q) Typhoon variable mode imager (Amersham Bioscience, cat. no. 63-0055-80) Vortex mixer (Vortex Genie-2, Mo Bio Laboratories, cat. no. 13111) Waters QToF premier mass spectrometer (Waters) and Masslynx software

    Purification:

    Article Title: Identification of a novel anoikis signalling pathway using the fungal virulence factor gliotoxin
    Article Snippet: The CNFy toxin (150 ng/ml) from Yersinia pseudotuberculosis and the C2I/C3 fusion toxin ( Clostridium botulinum and Clostridium Limosum , respectively) combined with the C2II toxin from Clostridium botulinum (collectively termed C3 toxin) was produced and purified as described , . .. The FAK inhibitor 14 (FAK14) was purchased from Tocris (Bristol, UK), and Cilengitide, a cyclic pentapeptide RGD compound (cyclo-[RGDfN(Me)V]) , the RGD peptide, DTT and iodocetamide were from Sigma (Taufkirchen, Germany).

    Lysis:

    Article Title: STIM2 drives Ca2+ oscillations through store-operated Ca2+ entry caused by mild store depletion
    Article Snippet: .. RBL-2H3 and HEK293 cells were harvested at the indicated time points after transfection, washed in PBS and lysed in 1 ml lysis buffer (in m m ): 75 NaCl, 40 NaF, 10 Iodocetamide, 50 Hepes, 10% IGEPAL, 1 phenylmethylsulphonyl fluoride and protease inhibitor cocktail (Sigma). .. The cell lysates from RBL-2H3 (see ) were precipitated with anti-STIM2(CT) rabbit polyclonal antibody (2.5 μg; Anaspec) for 1.5 h at 4°C and resolved by SDS-PAGE.

    Article Title: Site-specific chemical protein conjugation using genetically encoded aldehyde tags
    Article Snippet: .. Acetic acid (AcOH; Fisher Scientific, cat. no. A38-212) Acetic acid in deionized water (DI H2 O; 7.5%, vol/vol) Acetonitrile (ACN; Sigma-Aldrich, cat. no. 271004) Agar (Fisher Scientific, cat. no. BP2641) Alexa Fluor 647 C5-aminooxyacetamide (Molecular Probes, cat. no. ) Alexa Fluor 647 C5-aminooxyacetamide (10 mM in DMSO) Alkylation buffer Annealing buffer l -Arabinose (Sigma-Aldrich Chemical, cat. no. A3256) l -arabinose (20%, wt/vol) BamHI (NEB, cat. no. R0136) BglII (NEB, cat. no. R0104) Biomax 5K filter (Millipore, cat. no. UFV5BCC00) Bromophenol blue (Sigma-Aldrich Chemical, cat. no. B8026) Calf intestinal alkaline phosphatase (CIP; NEB, cat. no. M0290) Carbenicillin (Sigma-Aldrich Chemical, cat. no. C9231) CHO K1 cells (ATCC, cat. no. CCL-61) Complete EDTA-free protease inhibitor cocktail (Roche, cat. no. 11 873 580 001) Criterion XT MES running buffer (20×; Bio-Rad, cat. no. 161-0789) 1× Criterion XT MES running buffer Deionized water (DI H2 O) Denaturing buffer Dimethyl sulfoxide (DMSO; Sigma-Aldrich Chemical, cat. no. D8418) Dithiothreitol (DTT; Sigma-Aldrich Chemical, cat. no. 43815) DnaseI (NEB, cat. no. M0303) DpnI (NEB, cat. no. R0176) dNTPs (2.5 mM; Invitrogen, cat. no. ) EcoRV (NEB, cat. no. R0104) Elution buffer Ethylenediamine tetraacidic disodium salt (EDTA; Fisher Scientific, cat. no. BP120-1) Ex-Cell 325 PF CHO serum-free medium (Sigma-Aldrich, cat. no. 14340C) Fetal bovine serum (Mediatech, cat. no. 35-010-CV) l -Glutamine (Mediatech, cat. no. 25-005-CI) Glycerol (Sigma-Aldrich Chemical, cat. no. G5516) Glycerol (50%, vol/vol) Guanidine hydrochloride (Sigma-Aldrich, cat. no. G4505) HindIII (NEB, cat. no. R0104) Hydrochloric acid (HCl; EMD, cat. no. 3120-500) IgG elution buffer (Thermo Fisher Scientific, cat. no. 21004) Imidazole (Sigma-Aldrich Chemical, cat. no. I2399) Iodoacetamide (IAA; Sigma-Aldrich, cat. no. 068k53022) Isopropyl β- d -1-thiogalactopyranoside (IPTG; Sigma-Aldrich Chemical, cat. no. I5502) IPTG (isopropyl-1-thio-β- d -galactopyranoside, 1 M) Kanamycin (Sigma-Aldrich Chemical, cat. no. K1876) Labeling buffer (2×, 500 mM NaOAc, pH = 4.6) Lipofectamine 2000 (Invitrogen, cat. no. 11668019) Liquid nitrogen Lysis buffer Lysozyme (Sigma-Aldrich Chemical, cat. no. L6876) Methoxyamine hydrochloride (Sigma-Aldrich, cat. no. M1139) Ni-NTA agarose (Qiagen, cat. no. 30230) Oligonucleotides (see ) .. Amicon Ultra centrifugal filters (10 kDa molecular weight cutoff, Millipore, cat. no. UFC5010BK) Benchtop centrifuge (Sorvall Legend Mach 1.6 refrigerated, cat. no. 75004337) Centrifuge (Sorvall RC5C plus, cat. no. 74505) Criterion cell (Bio-Rad, cat. no. 165-6001) Criterion XT 4–12% (wt/vol) Bis-Tris gel (Bio-Rad, cat. no. 345-0123) Falcon tubes (15 ml; Fisher Scientific, cat. no. 05-527-90) Falcon tubes (50 ml, Fisher Scientific, cat. no. 14-432-22) Fast protein liquid chromatography (FPLC; GE Healthcare, ÄKTA purifier) HPLC (Waters) Microcentrifuge (Eppendorf 5417c, cat. no. 5417 000.315) Microcentrifuge tubes (2 ml; Fisher Scientific, cat. no. 02-681-291) NanoVue spectrophotometer (GE) PCR machine (Eppendorf, Mastercycler gradient, cat. no. 950000015) PCR tubes (Fisher Scientific, cat. no. 14-230-225) PD SpinTrap G-25 (GE Healthcare Life Sciences, cat. no. 28-9180-04) Reverse-phase HPLC column (Phenomenex, part: 00G-4396-B0) Shaking incubator (set at 220 r.p.m., refrigerated 4230; New Brunswick Scientific, cat. no. M1335-0012) Sterile 16-ml culture tubes (Fisher Scientific, cat. no. 05-540-6) Sterile 100 mm × 25 mm Petri dishes (Fisher Scientific, cat. no. 08-757-13) Sterile 0.22-μm polyethersulfone filters (Millipore, cat. no. SLGP033RS) Spin concentrator (Amicon Ultra 30kDa; Millipore, cat. no. UFC903008) Sonicator (Misonix sonicator 3000; Misonix) Tabletop rocking platform (Fisher Scientific, cat. no. 1404123Q) Typhoon variable mode imager (Amersham Bioscience, cat. no. 63-0055-80) Vortex mixer (Vortex Genie-2, Mo Bio Laboratories, cat. no. 13111) Waters QToF premier mass spectrometer (Waters) and Masslynx software

    Article Title: Prmt5 is a regulator of muscle stem cell expansion in adult mice
    Article Snippet: Mass spectrometry Muscle and satellite cell samples were homogenized in lysis buffer (4% SDS in 100 mm Tris/HCl, pH 7.6). .. Gel pieces were reduced with 10 mM dithiothreitol for 45 min at 56 °C and alkylated with 55 mM iodocetamide (BioUltra, Sigma-Aldrich) for 30 min at room temperature in the dark.

    Silver Staining:

    Article Title: Proteomic Analysis of Kidney Preservation Solutions Prior to Renal Transplantation
    Article Snippet: .. SDS, tris, urea, thiourea, ammonium bicarbonate, 3-[(3-cholamido propyl) dimethylamonio]-1-propanesulfonate (CHAPS), isopropanol, iodoacetamide, acrylamide, bisacrylamide, the ProteoSilver Plus Silver Stain Kit, glycerol, and α-cyanohydroxycinnamic acid were obtained from Sigma-Aldrich (St. Louis, MO, USA). .. Tetramethylethylenediamine, dithiothreitol (DTT), ethanol, and methanol were from Merck. (Whitehouse Station, NJ, USA).

    SDS Page:

    Article Title: STIM2 drives Ca2+ oscillations through store-operated Ca2+ entry caused by mild store depletion
    Article Snippet: RBL-2H3 and HEK293 cells were harvested at the indicated time points after transfection, washed in PBS and lysed in 1 ml lysis buffer (in m m ): 75 NaCl, 40 NaF, 10 Iodocetamide, 50 Hepes, 10% IGEPAL, 1 phenylmethylsulphonyl fluoride and protease inhibitor cocktail (Sigma). .. The cell lysates from RBL-2H3 (see ) were precipitated with anti-STIM2(CT) rabbit polyclonal antibody (2.5 μg; Anaspec) for 1.5 h at 4°C and resolved by SDS-PAGE.

    Article Title: Prmt5 is a regulator of muscle stem cell expansion in adult mice
    Article Snippet: Lysates were loaded on an SDS–PAGE (NuPAGE 4–12% BisTris gel, Invitrogen) and stained with colloidal Protein Staining Solution (Invitrogen). .. Gel pieces were reduced with 10 mM dithiothreitol for 45 min at 56 °C and alkylated with 55 mM iodocetamide (BioUltra, Sigma-Aldrich) for 30 min at room temperature in the dark.

    Produced:

    Article Title: Identification of a novel anoikis signalling pathway using the fungal virulence factor gliotoxin
    Article Snippet: The CNFy toxin (150 ng/ml) from Yersinia pseudotuberculosis and the C2I/C3 fusion toxin ( Clostridium botulinum and Clostridium Limosum , respectively) combined with the C2II toxin from Clostridium botulinum (collectively termed C3 toxin) was produced and purified as described , . .. The FAK inhibitor 14 (FAK14) was purchased from Tocris (Bristol, UK), and Cilengitide, a cyclic pentapeptide RGD compound (cyclo-[RGDfN(Me)V]) , the RGD peptide, DTT and iodocetamide were from Sigma (Taufkirchen, Germany).

    Concentration Assay:

    Article Title: An Effective Protocol for Proteome Analysis of Medaka (Oryzias latipes) after Acute Exposure to Ionizing Radiation
    Article Snippet: Materials 12 wild-type adult Medaka (~6–8 months old) with a body weight of 0.45 + 0.01 g and a 14 h:10 h light-dark cycle at 25 °C; 4 liter of filtered water; Anesthetic solution: 250 mg/L of tricaine methane sulfonate (MS-222) (Fisher, Fair Lawn, NJ, USA; cat: NC0342409), buffered with sodium bicarbonate at or near to neutral pH; Liquid nitrogen; Methanol HPLC grade (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: 34860-4L-R); Homogenizing solution kept at 4 °C: Mix chloroform (Fisher, Fair Lawn, NJ, USA; Cat. no.: C606SK-4), methanol, and Milli-Q water at ratio of 2:4:1.5, respectively; Acetone (Fisher, Fair Lawn, NJ, USA; Cat. no.: C606SK-4); Tris-HCl buffer: 1 mM Tris base (Fisher Scientific, Fair Lawn, NJ, USA; Cat. no.: BP152-500), pH 7.2 and 2% SDS (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: L4390); Pierce BCA protein assay kit (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: 23227); Bolt 4–12% Bis-Tris Plus gels (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: NW04120BOX); Protein ladder (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: LC5925); 2× Laemmli Sample Buffer (Bio-Rad, Hercules, CA, USA; Cat. no.: 1610737); Running buffer 1×: 50 mL of Bolt MOPS SDS running buffer (Thermo Scientific Pierce, Rockford, IL, USA; Cat. no.: B0001) mixed with 950 mL of deionized H2 O (dH2O); Plastic container 12 × 8 × 3 cm, rinsed previously with 50% aqueous methanol twice and one time with 100% methanol; Instant Blue Coomassie (Expedeon, Cambridge, UK; Cat. no: ISB1L); Destaining solution: 50 mL methanol, 5 mL acetic acid (J.T Baker, Center Valley, PA, USA; Cat. no.: 9508-33) and 50 mL dH2O; Isopropanol (Fisher, Fair Lawn, NJ, USA; Cat. no.: BP2635-4); Ambic solution: 0.158 g of ammonium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: AG141-500G) dissolved in 20 mL of dH2O to obtain a 100 mM solution; Acetonitrile HPLC grade (ACN) (Fisher, Fair Lawn, NJ, USA; Cat. no.: A988-4); 10 mM dithiothreitol (DTT): 0.015 g of DTT (Research Products International, Mount Prospect, IL, USA; Cat. no.: D11000-25) dissolved in 10 mL of Ambic solution; 55 mM iodoacetamide (IDA): 0.10 g of iodoacetamide (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: I1149-5G) dissolved in 10 mL of Ambic solution; Trypsin solution: 20 µg of trypsin (Promega, Madison, WI, USA; Cat. no.: V5111) in 1 mL of cold Ambic solution. .. Final concentration 20 ng/µL; Extraction solution: 50% ACN / 50% dH2O and 0.1% formic acid (Sigma-Aldrich, St. Louis, MO, USA; Cat. no.: F0507-500mL); Buffer A (0.1% aqueous formic acid); Buffer B (80% ACN, 0.1% aqueous formic acid).

    Liquid Chromatography with Mass Spectroscopy:

    Article Title: Prmt5 is a regulator of muscle stem cell expansion in adult mice
    Article Snippet: Gel pieces were reduced with 10 mM dithiothreitol for 45 min at 56 °C and alkylated with 55 mM iodocetamide (BioUltra, Sigma-Aldrich) for 30 min at room temperature in the dark. .. Released peptides were extracted (collecting separately the liquid mixture of each samples at each step) once by adding 100 μl of 30% acetonitrile liquid chromatography/mass spectrometry (LC/MS) grade (Thermo Scientific)/3% trifluoroacetic acid (Sigma-Aldrich) in water, and twice by adding 70% acetonitrile, followed by two final extractions with 100% acetonitrile.

    Staining:

    Article Title: Prmt5 is a regulator of muscle stem cell expansion in adult mice
    Article Snippet: Lysates were loaded on an SDS–PAGE (NuPAGE 4–12% BisTris gel, Invitrogen) and stained with colloidal Protein Staining Solution (Invitrogen). .. Gel pieces were reduced with 10 mM dithiothreitol for 45 min at 56 °C and alkylated with 55 mM iodocetamide (BioUltra, Sigma-Aldrich) for 30 min at room temperature in the dark.

    Electrofocusing:

    Article Title: Proteomic Analysis of Kidney Preservation Solutions Prior to Renal Transplantation
    Article Snippet: Materials The 2D electrophoresis equipment, i.e., isoelectric focusing (PROTEAN IEF) and Sodium dodecyl sulfate—polyacrylamide gel electrophoresis (protean II XI 2-D cell), the immobilized pH gradient (IPG; BioLyte) strips, mineral oil, glycine, ampholytes, and Ready Prep 2D clean-up kit were obtained from Bio-Rad (Hercules, CA, USA). .. SDS, tris, urea, thiourea, ammonium bicarbonate, 3-[(3-cholamido propyl) dimethylamonio]-1-propanesulfonate (CHAPS), isopropanol, iodoacetamide, acrylamide, bisacrylamide, the ProteoSilver Plus Silver Stain Kit, glycerol, and α-cyanohydroxycinnamic acid were obtained from Sigma-Aldrich (St. Louis, MO, USA).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    Millipore iodoacetamide
    Interaction of the bifunctional reagent DBB with H-Ras. (A) Modification of the K170-K185 peptide by DBB. The K170-K185 peptide was incubated with DBB, as described in the Methods section. When indicated the incubation mixture was subjected to reduction and alkylation with <t>iodoacetamide</t> to modify free cysteines. Results are representative of three independent assays. (B) Scheme of the cross-linking of two thiol groups by DBB. (C) Competition of 15d-PGJ 2 -B binding to H-Ras by DBB. H-Ras at 5 µM was incubated with 50 µM DBB for 30 min before addition of 1 µM 15d-PGJ 2 -B for 1 h. Incorporation of the biotinylated PG was assessed by SDS-PAGE, protein blot and detection with horseradish peroxidase (HRP)-streptavidin (upper panel). Incorporation of DBB (middle panel) and total H-Ras levels (lower panel) were assessed by fluorescence detection and western blot with anti-pan Ras antibody, respectively. (D) COS-7 cells transfected with the AU5-H-Ras vector were treated in the absence or presence of 50 µM DBB for 1 h. Aliquots from total cell lysates (10 µg of protein) were analyzed by SDS-PAGE. The upper component of the AU5-H-Ras doublet is indicated by an arrowhead. (E) COS-7 cells transfected with the AU5-H-Ras vector were treated with DBB and subjected to fractionation in Triton-X114 as above. In (D) and (E) levels of AU5-H-Ras were assessed by western blot with anti-AU5 antibody. The dotted line marks where lanes from the same gel have been cropped. (F) COS-7 cells transfected with YFP-RBD as above were treated with 50 µM DBB for 1 h before stimulation with 100 nM EGF, as indicated, and visualized live by confocal fluorescence microscopy. Dotted inserts show enlarged areas from the same cells for better visualization. Arrowheads mark the accumulation of YFP-RBD at defined locations of the cell periphery. Bar, 20 µm. Shown are representative images from three experiments with similar results.
    Iodoacetamide, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 311 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/iodoacetamide/product/Millipore
    Average 95 stars, based on 311 article reviews
    Price from $9.99 to $1999.99
    iodoacetamide - by Bioz Stars, 2020-02
    95/100 stars
      Buy from Supplier

    85
    Millipore iodoacetimide
    Interaction of the bifunctional reagent DBB with H-Ras. (A) Modification of the K170-K185 peptide by DBB. The K170-K185 peptide was incubated with DBB, as described in the Methods section. When indicated the incubation mixture was subjected to reduction and alkylation with <t>iodoacetamide</t> to modify free cysteines. Results are representative of three independent assays. (B) Scheme of the cross-linking of two thiol groups by DBB. (C) Competition of 15d-PGJ 2 -B binding to H-Ras by DBB. H-Ras at 5 µM was incubated with 50 µM DBB for 30 min before addition of 1 µM 15d-PGJ 2 -B for 1 h. Incorporation of the biotinylated PG was assessed by SDS-PAGE, protein blot and detection with horseradish peroxidase (HRP)-streptavidin (upper panel). Incorporation of DBB (middle panel) and total H-Ras levels (lower panel) were assessed by fluorescence detection and western blot with anti-pan Ras antibody, respectively. (D) COS-7 cells transfected with the AU5-H-Ras vector were treated in the absence or presence of 50 µM DBB for 1 h. Aliquots from total cell lysates (10 µg of protein) were analyzed by SDS-PAGE. The upper component of the AU5-H-Ras doublet is indicated by an arrowhead. (E) COS-7 cells transfected with the AU5-H-Ras vector were treated with DBB and subjected to fractionation in Triton-X114 as above. In (D) and (E) levels of AU5-H-Ras were assessed by western blot with anti-AU5 antibody. The dotted line marks where lanes from the same gel have been cropped. (F) COS-7 cells transfected with YFP-RBD as above were treated with 50 µM DBB for 1 h before stimulation with 100 nM EGF, as indicated, and visualized live by confocal fluorescence microscopy. Dotted inserts show enlarged areas from the same cells for better visualization. Arrowheads mark the accumulation of YFP-RBD at defined locations of the cell periphery. Bar, 20 µm. Shown are representative images from three experiments with similar results.
    Iodoacetimide, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/iodoacetimide/product/Millipore
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    iodoacetimide - by Bioz Stars, 2020-02
    85/100 stars
      Buy from Supplier


    Image Search Results


    Interaction of the bifunctional reagent DBB with H-Ras. (A) Modification of the K170-K185 peptide by DBB. The K170-K185 peptide was incubated with DBB, as described in the Methods section. When indicated the incubation mixture was subjected to reduction and alkylation with iodoacetamide to modify free cysteines. Results are representative of three independent assays. (B) Scheme of the cross-linking of two thiol groups by DBB. (C) Competition of 15d-PGJ 2 -B binding to H-Ras by DBB. H-Ras at 5 µM was incubated with 50 µM DBB for 30 min before addition of 1 µM 15d-PGJ 2 -B for 1 h. Incorporation of the biotinylated PG was assessed by SDS-PAGE, protein blot and detection with horseradish peroxidase (HRP)-streptavidin (upper panel). Incorporation of DBB (middle panel) and total H-Ras levels (lower panel) were assessed by fluorescence detection and western blot with anti-pan Ras antibody, respectively. (D) COS-7 cells transfected with the AU5-H-Ras vector were treated in the absence or presence of 50 µM DBB for 1 h. Aliquots from total cell lysates (10 µg of protein) were analyzed by SDS-PAGE. The upper component of the AU5-H-Ras doublet is indicated by an arrowhead. (E) COS-7 cells transfected with the AU5-H-Ras vector were treated with DBB and subjected to fractionation in Triton-X114 as above. In (D) and (E) levels of AU5-H-Ras were assessed by western blot with anti-AU5 antibody. The dotted line marks where lanes from the same gel have been cropped. (F) COS-7 cells transfected with YFP-RBD as above were treated with 50 µM DBB for 1 h before stimulation with 100 nM EGF, as indicated, and visualized live by confocal fluorescence microscopy. Dotted inserts show enlarged areas from the same cells for better visualization. Arrowheads mark the accumulation of YFP-RBD at defined locations of the cell periphery. Bar, 20 µm. Shown are representative images from three experiments with similar results.

    Journal: PLoS ONE

    Article Title: The C-Terminus of H-Ras as a Target for the Covalent Binding of Reactive Compounds Modulating Ras-Dependent Pathways

    doi: 10.1371/journal.pone.0015866

    Figure Lengend Snippet: Interaction of the bifunctional reagent DBB with H-Ras. (A) Modification of the K170-K185 peptide by DBB. The K170-K185 peptide was incubated with DBB, as described in the Methods section. When indicated the incubation mixture was subjected to reduction and alkylation with iodoacetamide to modify free cysteines. Results are representative of three independent assays. (B) Scheme of the cross-linking of two thiol groups by DBB. (C) Competition of 15d-PGJ 2 -B binding to H-Ras by DBB. H-Ras at 5 µM was incubated with 50 µM DBB for 30 min before addition of 1 µM 15d-PGJ 2 -B for 1 h. Incorporation of the biotinylated PG was assessed by SDS-PAGE, protein blot and detection with horseradish peroxidase (HRP)-streptavidin (upper panel). Incorporation of DBB (middle panel) and total H-Ras levels (lower panel) were assessed by fluorescence detection and western blot with anti-pan Ras antibody, respectively. (D) COS-7 cells transfected with the AU5-H-Ras vector were treated in the absence or presence of 50 µM DBB for 1 h. Aliquots from total cell lysates (10 µg of protein) were analyzed by SDS-PAGE. The upper component of the AU5-H-Ras doublet is indicated by an arrowhead. (E) COS-7 cells transfected with the AU5-H-Ras vector were treated with DBB and subjected to fractionation in Triton-X114 as above. In (D) and (E) levels of AU5-H-Ras were assessed by western blot with anti-AU5 antibody. The dotted line marks where lanes from the same gel have been cropped. (F) COS-7 cells transfected with YFP-RBD as above were treated with 50 µM DBB for 1 h before stimulation with 100 nM EGF, as indicated, and visualized live by confocal fluorescence microscopy. Dotted inserts show enlarged areas from the same cells for better visualization. Arrowheads mark the accumulation of YFP-RBD at defined locations of the cell periphery. Bar, 20 µm. Shown are representative images from three experiments with similar results.

    Article Snippet: When indicated, aliquots of H-Ras, tryptic digests or K170-K185 peptide were subsequently incubated with 10 mM DTT for 30 min followed by 50 mM iodoacetamide for 30 min at r.t. before purification on ZipTip C18 (Millipore) and MALDI-TOF MS analysis on an AUTOFLEX III MALDI-TOF-TOF instrument (Bruker-Franzen Analytik, Bremen, FRG) operated in the positive mode, as reported in detail .

    Techniques: Modification, Incubation, Binding Assay, SDS Page, Fluorescence, Western Blot, Transfection, Plasmid Preparation, Fractionation, Microscopy

    Effect of reducing agents on PAO modification of the K170-K185 peptide. (A) In the upper panels, the K170-K185 peptide was incubated with PAO and analyzed by MALDI-TOF MS. In the right panel, the incubation mixture was treated with 10 mM DTT and subsequently with 50 mM iodoacetamide. In the lower panels, the K170-K185 peptide was incubated with PAO in the presence of the indicated concentrations of GSH before analysis by MALDI-TOF MS. (B) Summary of the theoretical m/z of peptides for which compatible peaks were observed experimentally in the assays shown in (A).

    Journal: PLoS ONE

    Article Title: The C-Terminus of H-Ras as a Target for the Covalent Binding of Reactive Compounds Modulating Ras-Dependent Pathways

    doi: 10.1371/journal.pone.0015866

    Figure Lengend Snippet: Effect of reducing agents on PAO modification of the K170-K185 peptide. (A) In the upper panels, the K170-K185 peptide was incubated with PAO and analyzed by MALDI-TOF MS. In the right panel, the incubation mixture was treated with 10 mM DTT and subsequently with 50 mM iodoacetamide. In the lower panels, the K170-K185 peptide was incubated with PAO in the presence of the indicated concentrations of GSH before analysis by MALDI-TOF MS. (B) Summary of the theoretical m/z of peptides for which compatible peaks were observed experimentally in the assays shown in (A).

    Article Snippet: When indicated, aliquots of H-Ras, tryptic digests or K170-K185 peptide were subsequently incubated with 10 mM DTT for 30 min followed by 50 mM iodoacetamide for 30 min at r.t. before purification on ZipTip C18 (Millipore) and MALDI-TOF MS analysis on an AUTOFLEX III MALDI-TOF-TOF instrument (Bruker-Franzen Analytik, Bremen, FRG) operated in the positive mode, as reported in detail .

    Techniques: Modification, Incubation, Mass Spectrometry

    Modification of the K170-K185 peptide from the C-terminal region of H-Ras by cyPG. (A) The synthetic K170-K185 peptide was incubated with the indicated cyPG and the resulting adducts analyzed by MALDI-TOF MS. When indicated, incubation mixtures were treated with 50 mM iodoacetamide after addition of 10 mM DTT. Spectra presented are representative from at least three independent assays per experimental condition. Structures of the cyPG used are shown in insets. Electrophilic carbons are marked by asterisks. (B) Summary of the theoretical peptide adducts for which compatible peaks were observed experimentally. +CAM indicates that the peak is compatible with the formation of carbamidomethyl cysteine subsequent to iodoacetamide treatment. (C) Ribbon diagram for the theoretical backbone structure of the H-Ras K170-K185 peptide (in green) modified by addition of 15d-PGJ 2 to cysteines 181 and 184. The side-chains of the cysteine residues and the 15d-PGJ 2 ligand are displayed in yellow and a default atom-type color scheme, respectively.

    Journal: PLoS ONE

    Article Title: The C-Terminus of H-Ras as a Target for the Covalent Binding of Reactive Compounds Modulating Ras-Dependent Pathways

    doi: 10.1371/journal.pone.0015866

    Figure Lengend Snippet: Modification of the K170-K185 peptide from the C-terminal region of H-Ras by cyPG. (A) The synthetic K170-K185 peptide was incubated with the indicated cyPG and the resulting adducts analyzed by MALDI-TOF MS. When indicated, incubation mixtures were treated with 50 mM iodoacetamide after addition of 10 mM DTT. Spectra presented are representative from at least three independent assays per experimental condition. Structures of the cyPG used are shown in insets. Electrophilic carbons are marked by asterisks. (B) Summary of the theoretical peptide adducts for which compatible peaks were observed experimentally. +CAM indicates that the peak is compatible with the formation of carbamidomethyl cysteine subsequent to iodoacetamide treatment. (C) Ribbon diagram for the theoretical backbone structure of the H-Ras K170-K185 peptide (in green) modified by addition of 15d-PGJ 2 to cysteines 181 and 184. The side-chains of the cysteine residues and the 15d-PGJ 2 ligand are displayed in yellow and a default atom-type color scheme, respectively.

    Article Snippet: When indicated, aliquots of H-Ras, tryptic digests or K170-K185 peptide were subsequently incubated with 10 mM DTT for 30 min followed by 50 mM iodoacetamide for 30 min at r.t. before purification on ZipTip C18 (Millipore) and MALDI-TOF MS analysis on an AUTOFLEX III MALDI-TOF-TOF instrument (Bruker-Franzen Analytik, Bremen, FRG) operated in the positive mode, as reported in detail .

    Techniques: Modification, Incubation, Mass Spectrometry, Chick Chorioallantoic Membrane Assay

    Representative HPLC spectra of pure IAA and IAM and of 15-h P. chlororaphis O6 ( Pc O6) cell cultures, indicating the presence of IAM as an intermediate of IAA production in the presence of trp. Inset: a representative TLC-based assay for direct IAA production

    Journal: Applied and Environmental Microbiology

    Article Title: Production of Indole-3-Acetic Acid via the Indole-3-Acetamide Pathway in the Plant-Beneficial Bacterium Pseudomonas chlororaphis O6 Is Inhibited by ZnO Nanoparticles but Enhanced by CuO Nanoparticles

    doi: 10.1128/AEM.07424-11

    Figure Lengend Snippet: Representative HPLC spectra of pure IAA and IAM and of 15-h P. chlororaphis O6 ( Pc O6) cell cultures, indicating the presence of IAM as an intermediate of IAA production in the presence of trp. Inset: a representative TLC-based assay for direct IAA production

    Article Snippet: Both IAM and IAA were detected by thin-layer chromatography (TLC) on plates of silica Gel 60 (EMD Chemicals Inc., Darmstadt, Germany).

    Techniques: High Performance Liquid Chromatography, Thin Layer Chromatography

    Cysteines involved in disulfide bridges, as shown by secondly modified with iodoacetamide. Reflectron MALDI-TOF MS spectrum of one HPLC fraction from the combination of trypsin, Asp-N, and Glu-C triple-enzyme digest of OMC demonstrates modified cysteine

    Journal:

    Article Title: Identification of surface-exposed components of MOMP of Chlamydia trachomatis serovar F

    doi: 10.1110/ps.051616206

    Figure Lengend Snippet: Cysteines involved in disulfide bridges, as shown by secondly modified with iodoacetamide. Reflectron MALDI-TOF MS spectrum of one HPLC fraction from the combination of trypsin, Asp-N, and Glu-C triple-enzyme digest of OMC demonstrates modified cysteine

    Article Snippet: The nonreduced proteolytic OMC digests were pyridylethylated with 1 M VP before RP-HPLC separation, half of each RP-HPLC collected fraction was reduced with 20 mM DTT and carboxymethylated again with 100 mM iodoacetamide (IAM) ( ) in darkness for half an hour before C18 Ziptip (Millipore) cleanup.

    Techniques: Modification, Mass Spectrometry, High Performance Liquid Chromatography