C18 cartridges were primed using 70% acetonitrile (ACN) with 0.1% formic acid (FA) (100 μl, 300 μl/min), equilibrated in 2% ACN with 0.1% FA (50 μl, 5 μl/min), and eluates described above were loaded (280 μl, 5 μl/min). For performing reduction and alkylation, peptides were reduced in HEPES buffer containing 5 mM Tris (2-carboxyethyl) phosphine (TCEP; 100 μl, 5 μl/min), alkylated with 40 mM IAA (100 μl, 2 μl/min), and washed with HEPES buffer/5 mM TCEP (100 μl, 10 μl/min) followed by HEPES buffer alone (100 μl, 10 μl/min). For performing oxidation, the oxidant solution (5% formic acid, 1X H2O2) was prepared by mixing (1:1) 10% formic acid with 2X H2O2 (concentration varies). H2O2 was diluted with water from fresh 30% H2O2 (Sigma) to the appropriate concentration. Peptides were then washed with this oxidant solution (150 μl, 5 μl/min) followed by HEPES buffer (100 μl, 10 μl/min). For TMT tagging, 85 μg of TMT6plex reagent (Thermo Fisher) in HEPES containing 8% acetonitrile (ACN) was loaded over 25 min (50 μl at 2 μl/min). Finally, peptides were washed with 2% ACN with 0.1% FA (100 μl, 10 μl/min) and eluted in 30% ACN with 0.1% FA (50 μl, 5 μl/min). Fractions were transferred to glass LCMS vials (Agilent), dried for 10 min by speedvac, and stored at −80 °C.
Glass lcms vials
Manufactured by Agilent Technologies
Glass LCMS vials are laboratory containers designed for use in liquid chromatography-mass spectrometry (LCMS) analysis. They provide a secure and inert storage environment for samples during LCMS testing. The vials are made of high-quality borosilicate glass, ensuring sample integrity and compatibility with LCMS instrumentation.
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Lab products found in correlation
2 protocols using glass lcms vials
1
Peptide Purification and Modification Protocol
2
Peptide Purification and Labeling Protocol
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C18 cartridges were primed using 70% acetonitrile (ACN) with 0.1% formic acid (FA) (100 µL, 300 µL/min), equilibrated in 2% ACN with 0.1% FA (50 µL, 5 µL/min), and eluates described above were loaded (280 µL, 5 µL/min). For performing reduction and alkylation, peptides were reduced in HEPES buffer containing 5 mM Tris (2-carboxyethyl) phosphine (TCEP; 100 µL, 5 µL/min), alkylated with 40 mM IAA (100 µL, 2 µL/min), and washed with HEPES buffer/5 mM TCEP (100 µL, 10 µL/min) followed by HEPES buffer alone (100 µL, 10 µL/min). For performing oxidation, the oxidant solution (5% formic acid, 1X H 2 O 2 ) was prepared by mixing (1:1) 10% formic acid with 2X H 2 O 2 (concentration varies). H 2 O 2 was diluted with water from fresh 30% H 2 O 2 (Sigma) to the appropriate concentration.
Peptides were then washed with this oxidant solution (150 µL, 5 µL/min) followed by HEPES buffer (100 µL, 10 µL/min). For TMT tagging, 85 µg of TMT6plex reagent (ThermoFisher) in HEPES containing 8% acetonitrile (ACN) was loaded over 25 min (50 µL at 2 µL/min). Finally, peptides were washed with 2% ACN with 0.1% FA (100 µL, 10 µL/min) and eluted in 30% ACN with 0.1% FA (50 µL, 5 µL/min). Fractions were transferred to glass LCMS vials (Agilent), dried for 10 min by speedvac and stored at -80°C.
Peptides were then washed with this oxidant solution (150 µL, 5 µL/min) followed by HEPES buffer (100 µL, 10 µL/min). For TMT tagging, 85 µg of TMT6plex reagent (ThermoFisher) in HEPES containing 8% acetonitrile (ACN) was loaded over 25 min (50 µL at 2 µL/min). Finally, peptides were washed with 2% ACN with 0.1% FA (100 µL, 10 µL/min) and eluted in 30% ACN with 0.1% FA (50 µL, 5 µL/min). Fractions were transferred to glass LCMS vials (Agilent), dried for 10 min by speedvac and stored at -80°C.
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