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  • 99
    Thermo Fisher acetonitrile
    Acetonitrile, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1491 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    acetonitrile - by Bioz Stars, 2020-09
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    99
    Millipore acetonitrile acn
    Acetonitrile Acn, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1208 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Fisher Scientific acetonitrile acn
    Chromatographic separation of (A) 18 SPMs (100 nM each) and (B) 5 deuterated internal standards (20 nM) covered by the method including (C) DHA derived D-series resolvins, (D) EPA derived E-series resolvins, (E) EPA and ARA derived lipoxins, DHA derived (F) protectins, and (G) maresins. Separation was carried out on an RP-18 column (2.1 × 150 mm, particle size 1.8 μm, pore size 9.5 nm) with a H 2 <t>O/MeOH/ACN/HAc</t> gradient.
    Acetonitrile Acn, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 93/100, based on 719 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Avantor methanol meoh
    Chromatographic separation of (A) 18 SPMs (100 nM each) and (B) 5 deuterated internal standards (20 nM) covered by the method including (C) DHA derived D-series resolvins, (D) EPA derived E-series resolvins, (E) EPA and ARA derived lipoxins, DHA derived (F) protectins, and (G) maresins. Separation was carried out on an RP-18 column (2.1 × 150 mm, particle size 1.8 μm, pore size 9.5 nm) with a H 2 <t>O/MeOH/ACN/HAc</t> gradient.
    Methanol Meoh, supplied by Avantor, used in various techniques. Bioz Stars score: 93/100, based on 239 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    methanol meoh - by Bioz Stars, 2020-09
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    89
    Phenomenex aeris peptide xb c18 column
    Chromatographic separation of (A) 18 SPMs (100 nM each) and (B) 5 deuterated internal standards (20 nM) covered by the method including (C) DHA derived D-series resolvins, (D) EPA derived E-series resolvins, (E) EPA and ARA derived lipoxins, DHA derived (F) protectins, and (G) maresins. Separation was carried out on an RP-18 column (2.1 × 150 mm, particle size 1.8 μm, pore size 9.5 nm) with a H 2 <t>O/MeOH/ACN/HAc</t> gradient.
    Aeris Peptide Xb C18 Column, supplied by Phenomenex, used in various techniques. Bioz Stars score: 89/100, based on 160 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Fisher Scientific methanol meoh
    Chromatographic separation of (A) 18 SPMs (100 nM each) and (B) 5 deuterated internal standards (20 nM) covered by the method including (C) DHA derived D-series resolvins, (D) EPA derived E-series resolvins, (E) EPA and ARA derived lipoxins, DHA derived (F) protectins, and (G) maresins. Separation was carried out on an RP-18 column (2.1 × 150 mm, particle size 1.8 μm, pore size 9.5 nm) with a H 2 <t>O/MeOH/ACN/HAc</t> gradient.
    Methanol Meoh, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 92/100, based on 480 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore ambic solution
    Chromatographic separation of (A) 18 SPMs (100 nM each) and (B) 5 deuterated internal standards (20 nM) covered by the method including (C) DHA derived D-series resolvins, (D) EPA derived E-series resolvins, (E) EPA and ARA derived lipoxins, DHA derived (F) protectins, and (G) maresins. Separation was carried out on an RP-18 column (2.1 × 150 mm, particle size 1.8 μm, pore size 9.5 nm) with a H 2 <t>O/MeOH/ACN/HAc</t> gradient.
    Ambic Solution, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Promega sequencing grade modified trypsin
    Chromatographic separation of (A) 18 SPMs (100 nM each) and (B) 5 deuterated internal standards (20 nM) covered by the method including (C) DHA derived D-series resolvins, (D) EPA derived E-series resolvins, (E) EPA and ARA derived lipoxins, DHA derived (F) protectins, and (G) maresins. Separation was carried out on an RP-18 column (2.1 × 150 mm, particle size 1.8 μm, pore size 9.5 nm) with a H 2 <t>O/MeOH/ACN/HAc</t> gradient.
    Sequencing Grade Modified Trypsin, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 3265 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Burdick & Jackson acetonitrile acn
    Chromatographic separation of (A) 18 SPMs (100 nM each) and (B) 5 deuterated internal standards (20 nM) covered by the method including (C) DHA derived D-series resolvins, (D) EPA derived E-series resolvins, (E) EPA and ARA derived lipoxins, DHA derived (F) protectins, and (G) maresins. Separation was carried out on an RP-18 column (2.1 × 150 mm, particle size 1.8 μm, pore size 9.5 nm) with a H 2 <t>O/MeOH/ACN/HAc</t> gradient.
    Acetonitrile Acn, supplied by Burdick & Jackson, used in various techniques. Bioz Stars score: 93/100, based on 188 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    honeywell international methanol meoh
    Chromatographic separation of (A) 18 SPMs (100 nM each) and (B) 5 deuterated internal standards (20 nM) covered by the method including (C) DHA derived D-series resolvins, (D) EPA derived E-series resolvins, (E) EPA and ARA derived lipoxins, DHA derived (F) protectins, and (G) maresins. Separation was carried out on an RP-18 column (2.1 × 150 mm, particle size 1.8 μm, pore size 9.5 nm) with a H 2 <t>O/MeOH/ACN/HAc</t> gradient.
    Methanol Meoh, supplied by honeywell international, used in various techniques. Bioz Stars score: 91/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore hplc grade
    Chromatographic separation of (A) 18 SPMs (100 nM each) and (B) 5 deuterated internal standards (20 nM) covered by the method including (C) DHA derived D-series resolvins, (D) EPA derived E-series resolvins, (E) EPA and ARA derived lipoxins, DHA derived (F) protectins, and (G) maresins. Separation was carried out on an RP-18 column (2.1 × 150 mm, particle size 1.8 μm, pore size 9.5 nm) with a H 2 <t>O/MeOH/ACN/HAc</t> gradient.
    Hplc Grade, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2238 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore iodoacetamide
    Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM <t>iodoacetamide</t> for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.
    Iodoacetamide, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 12698 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher lc ms grade methanol
    Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM <t>iodoacetamide</t> for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.
    Lc Ms Grade Methanol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Avantor lc ms grade water
    Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM <t>iodoacetamide</t> for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.
    Lc Ms Grade Water, supplied by Avantor, used in various techniques. Bioz Stars score: 92/100, based on 139 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    lc ms grade water - by Bioz Stars, 2020-09
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    99
    Millipore formic acid
    Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM <t>iodoacetamide</t> for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.
    Formic Acid, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1599 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Avantor acetonitrile
    Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM <t>iodoacetamide</t> for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.
    Acetonitrile, supplied by Avantor, used in various techniques. Bioz Stars score: 92/100, based on 1317 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Fisher Scientific acetonitrile
    Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM <t>iodoacetamide</t> for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.
    Acetonitrile, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 94/100, based on 2215 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher edta
    Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM <t>iodoacetamide</t> for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.
    Edta, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 16222 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore hydrochloric acid hcl
    Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM <t>iodoacetamide</t> for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.
    Hydrochloric Acid Hcl, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 858 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore ammonium hydroxide solution
    Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM <t>iodoacetamide</t> for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.
    Ammonium Hydroxide Solution, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 332 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Agilent technologies 1200 series lc ms
    Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM <t>iodoacetamide</t> for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.
    1200 Series Lc Ms, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 88/100, based on 91 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Agilent technologies 1220 infinity lc
    Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM <t>iodoacetamide</t> for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.
    1220 Infinity Lc, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 91/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Agilent technologies 1260 infinity bio inert lc system
    Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM <t>iodoacetamide</t> for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.
    1260 Infinity Bio Inert Lc System, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 95/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Chromatographic separation of (A) 18 SPMs (100 nM each) and (B) 5 deuterated internal standards (20 nM) covered by the method including (C) DHA derived D-series resolvins, (D) EPA derived E-series resolvins, (E) EPA and ARA derived lipoxins, DHA derived (F) protectins, and (G) maresins. Separation was carried out on an RP-18 column (2.1 × 150 mm, particle size 1.8 μm, pore size 9.5 nm) with a H 2 O/MeOH/ACN/HAc gradient.

    Journal: Frontiers in Pharmacology

    Article Title: Development of an Optimized LC-MS Method for the Detection of Specialized Pro-Resolving Mediators in Biological Samples

    doi: 10.3389/fphar.2019.00169

    Figure Lengend Snippet: Chromatographic separation of (A) 18 SPMs (100 nM each) and (B) 5 deuterated internal standards (20 nM) covered by the method including (C) DHA derived D-series resolvins, (D) EPA derived E-series resolvins, (E) EPA and ARA derived lipoxins, DHA derived (F) protectins, and (G) maresins. Separation was carried out on an RP-18 column (2.1 × 150 mm, particle size 1.8 μm, pore size 9.5 nm) with a H 2 O/MeOH/ACN/HAc gradient.

    Article Snippet: Acetonitrile (ACN), LC-MS grade methanol (MeOH) and acetic acid were obtained from Fisher Scientific (Schwerte, Germany).

    Techniques: Derivative Assay, Acetylene Reduction Assay, HAC Assay

    Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM iodoacetamide for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.

    Journal: Autophagy

    Article Title: Phosphoproteome-based kinase activity profiling reveals the critical role of MAP2K2 and PLK1 in neuronal autophagy

    doi: 10.1080/15548627.2017.1371393

    Figure Lengend Snippet: Flowchart of the identification of the quantitative phosphoproteome in neuronal autophagy. According to the weights of the labeled essential amino acids, N2a cells were divided into 3 groups, including “heavy” ( 13 C 6 15 N 2 -Lysine, 13 C 6 15 N 4 -Arginine), “middle” ( 13 C 4 14 N 2 -Lysine, 13 C 6 14 N 4 -Arginine) and “light” ( 12 C 6 14 N 2 -Lysine, 12 C 6 14 N 4 -Arginine). After checking the labeling efficiency ( > 96%), the “light” and “middle” groups were treated with 15 μM Cory or Cory B for 3 h (Fig. S1), and the “heavy” group, which acted as a control group, was treated with 0.1% (v:v) DMSO. The cell lysates were mixed at a ratio of 1:1:1 and reduced with 10 mM dithiothreitol for 1 h at 56°C and then alkylated with 20 mM iodoacetamide for 45 min at room temperature and protected from light. Then, proteins were digested in solution with trypsin gold at a ratio of 1:50 (trypsin to protein) overnight and 1:100 (trypsin to protein) for another 4 h. After fractionation by high-pH reverse-phase HPLC, peptides were divided into 12 fractions. Finally, enriched phosphopeptides were subjected to LC-MS/MS for the identification of the quantitative phosphoproteome.

    Article Snippet: H2 O was purchased from Thermo (10977023); ACN (acetonitrile; A998–1) and fa (ethyl alcohol; A995–4) was purchased from Fisher Chemical; formic acid (FA; 5.43804) was purchased from Fluka; DMEM medium (11965118) and fetal bovine serum (FBS; 16000044) were purchased from Invitrogen; SILAC™ Protein Identification and Quantitation Media Kit was purchased from Thermo (SP10001); Sequencing Grade Modified Trypsin was purchased from Promega (V5280); trifluoroacetic acid (TFA; 574732), iodoacetamide (A3221) and dithiothreitol (43817) were purchased from Sigma; 2-D Quant kit was purchased from GE Healthcare (80–6483–56); IMAC beads was purchased from Dalian Institute of Chemical Physics.

    Techniques: Labeling, Fractionation, High Performance Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry