The chemical reagents acetonitrile, ethanol, methanol, acetone, ammonium formate (high-performance liquid chromatography (HPLC) grade), and trifluoroacetic acid (TFA) were obtained from Sigma Corporation (Sigma, USA) and Fisher Science Corporation (Thermo, USA). The ultrapure-grade water, utilized for the HPLC and subsequent tandem mass spectrometry (MS/MS) analysis procedures, was generated from the MilliQ (Millipore, USA)-type water. All iTRAQ reagents and buffers were obtained from Applied Biosystems (Applied Biosystems, Foster City, CA, USA).
Itraq reagents and buffers
Manufactured by Thermo Fisher Scientific
Sourced in United States
iTRAQ reagents and buffers are a set of chemical compounds used in mass spectrometry-based proteomics for the relative and absolute quantification of proteins in different samples. The core function of these reagents is to enable the labeling and identification of peptides derived from protein samples.
Automatically generated - may contain errors
Lab products found in correlation
Acetone, by Merck Group (2 mentions)
Ethanol, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Trifluoroacetic acid, by Merck Group (1 mentions)
Ammonium formate, by Merck Group (1 mentions)
Protein assay kit, by Bio-Rad (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Transwell chamber, by Corning (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Pkh67 membrane dye, by Merck Group (1 mentions)
5 n n dimethyl amiloride hydrochloride, by Merck Group (1 mentions)
Trifluoracetic acid tfa, by Merck Group (1 mentions)
Hplc grade methanol, by Merck Group (1 mentions)
Proteomics grade trypsin, by Merck Group (1 mentions)
Ammonium bicarbonate, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Hplc grade water, by Merck Group (1 mentions)
α cyano 4 hydroxy trans cynnamic acid α chca, by Merck Group (1 mentions)
4 protocols using itraq reagents and buffers
1
Quantitative Mass Spectrometry Reagents
2
Protein Assay and RNA Isolation
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Protein Assay Kit was purchased from Bio-Rad (Hercules, CA, USA). Reagent for total RNA isolation was from Qiagen (Valencia, CA, USA). Reagents for qPCR were obtained from (Takara, Dalian, China). All iTRAQ reagents and buffers were obtained from Applied Biosystems Inc. (ABI, Foster City, CA). The rest of reagent grade chemicals used in the present study were from Sigma Aldrich (St. Louis, MO, USA) or Fisher Scientific (Pittsburg, PA, USA).
3
Optimized Cell Migration Assay
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RPMI 1640, fetal bovine serum (FBS), proteases, proteases inhibitors, and antibiotics were purchased from Gibco-BRL (Shanghai, China). PKH67 membrane dye and 5-(N, N-Dimethyl) amiloride hydrochloride (DMA) were purchased from Sigma-Aldrich (MO, USA). Trans-well chambers were purchased from Corning Life Sciences (MA, USA). All iTRAQ reagents and buffers were purchased from Applied Biosystems, Inc. (Foster City, CA). All other reagents were of the highest analytical grades available and unless otherwise stated were purchased from Sigma-Aldrich (MO, USA).
4
Quantitative Proteomics Workflow
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Ammonium Bicarbonate (NH4HCO3, 99.5%), trypsin (proteomics grade), α-cyano-4-hydroxy-transcynnamic acid (α-CHCA, 99,0%), water (HPLC grade), trifluoracetic acid (TFA, 99,0%), methanol (HPLC grade), acetone, protease inhibitor cocktail and protein standards for protein molecular weight marker were purchased from Fluka-Sigma Aldrich S.r.l. (Milan, Italy). Protein standards and reagent for protein quantification were acquired by Bio-Rad's Laboratories, Inc. (Monza, Italy). iTRAQ reagents and buffers were obtained from Applied Biosystems (Foster City, CA). Peptide and protein standards, for mass spectrometer external calibration, were prepared from the Sequazime peptide mass standard kit (Applied Biosystems, Framingham, MA, USA).
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