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To study the acetylation of human serum albumin (HSA) in presence and absence of Chlorpyrifos-oxon using 2-Naphthyl Acetate (2NA) as substrate and digestion with Trypsin

ABSTRACT: This is study done for academic purpose and funded by ICMR, New Delhi, India, sanctioned project titled- To study the acetylation of human serum albumin (HSA) in presence and absence of Chlorpyrifos-oxon using 2-Naphthyl Acetate (2NA) as substrate and digestion with Trypsin. Project File No. 5/4/1-25/2020-NCD-1. Sample processing protocol: For this, 1 ml of 1 mg/ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with Chlorpyrifos-oxon (in Acetonitrile)(working concentration of 5 microMolar for 1 hour at 37 degree Celsius. Further, acetylation was done using 2NA (prepared in Acetonitrile) for 45 minutes at 37 degree Celsius. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 minutes followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree Celsius. These were then subjected to Trypsin digestion (2 microgram, Promega, MS grade).Next, samples to be digested by Trypsin were incubated for overnight at 37 degree Celsius. Trypsin digestion was stopped by adding 1 percent Trifluoroacetic acid solution. The sample clean-up with C18 resins column was performed and the aliquots were then sent for data acquisition. Data Processing protocol: The MaxQuant software (2.0.3.1) was used for the analysis of data. Under group-specific parameters Acetyl (Y), Acetyl K, Acetyl (N-term), and (Acetyl protein N-term) were selected as variable modifications whereas the Carbamidomethyl (C) was considered a fixed modification. The label-free quantification was done and the albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. For digestion, Trypsin enzyme was added to MaxQuant enzyme file. The albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. Description of the raw files is as follow. Chlorpyrifos-oxon is Human serum albumin treated with Chlorpyrifos-oxon and Chlorpyrifos-oxon _2NA is Human serum Albumin treated with Chlorpyrifos-oxon and then substrate (2NA) was added.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Dibyajyoti Banerjee

PROVIDER: MSV000100233 | MassIVE | Mon Dec 15 23:55:00 GMT 2025

REPOSITORIES: MassIVE

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Project description:This is study done for academic purpose and funded by ICMR, New Delhi, India, sanctioned project titled:- To study the acetylation of human serum albumin (HSA) in presence and absence of Chlorpyrifos using 2-Naphthyl Acetate (2NA) as substrate and digestion with Trypsin. Project File No. 5/4/1-25/2020-NCD-1. Sample processing protocol: For this, 1 ml of 1 mg/ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with Chlorpyrifos (in Acetonitrile)(working concentration of 5 microMolar for 1 hour at 37 degree Celsius. Further, acetylation was done using 2NA (prepared in Acetonitrile) for 45 minutes at 37 degree Celsius. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 minutes followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree Celsius. These were then subjected to Trypsin digestion (2 microgram, Promega, MS grade).Next, samples to be digested by Trypsin were incubated for overnight at 37 degree Celsius. Trypsin digestion was stopped by adding 1 percent Trifluoroacetic acid solution. The sample clean-up with C18 resins column was performed and the aliquots were then sent for data acquisition. Data Processing protocol: The MaxQuant software (2.0.3.1) was used for the analysis of data. Under group-specific parameters Acetyl (Y), Acetyl K, Acetyl (N-term), and (Acetyl protein N-term) were selected as variable modifications whereas the Carbamidomethyl (C) was considered a fixed modification. The label-free quantification was done and the albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. For digestion, Trypsin enzyme was added to MaxQuant enzyme file. The albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. Description of the raw files is as follow. Chlorpyrifos is Human serum albumin treated with Chlorpyrifos and Chlorpyrifos _2NA is Human serum Albumin treated with Chlorpyrifos and then substrate (2NA) was added.

Project description:This is study done for academic purpose and funded by ICMR, New Delhi, India, sanctioned project titled:- To study the acetylation of human serum albumin (HSA) in presence and absence of Chlorpyrifos using 2-Naphthyl Acetate (2NA) as substrate and digestion with Pepsin. Project File No. 5/4/1-25/2020-NCD-1. Sample processing protocol: For this, 1 ml of 1 mg/ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with Chlorpyrifos (in Acetonitrile)(working concentration of 5 microMolar for 1 hour at 37 degree Celsius. Further, acetylation was done using 2NA (prepared in Acetonitrile) for 45 minutes at 37 degree Celsius. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 minutes followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree Celsius. These were then subjected to Pepsin digestion (2 microgram, Promega, MS grade, prepared in acidic water).Next, samples to be digested by pepsin were incubated for 2 hours at 37 degree Celsius. Pepsin digestion was stopped by heating samples at 95 degree Celsius. The sample clean-up with C18 resins column was performed and the aliquots were then sent for data acquisition. Data Processing protocol: The MaxQuant software (2.0.3.1) was used for the analysis of data. Under group-specific parameters acetyl K, acetyl (N-term), and (acetyl protein N-term) were selected as variable modifications whereas the Carbamidomethyl (C) was considered a fixed modification. The label-free quantification was done and the albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. For digestion, Pepsin enzyme was added to MaxQuant enzyme file. The albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. Description of the raw files is as follow. HSA_Chlorpyrifos is Human serum albumin treated with Chlorpyrifos and HSA_Chlorpyrifos _2NA is Human serum Albumin treated with Chlorpyrifos and then substrate (2NA) was added.

Project description:This is study done for academic purpose and funded by ICMR, New Delhi, India, sanctioned project titled- To study the acetylation of human serum albumin (HSA) in presence and absence of Carbaryl using 2-Naphthyl Acetate (2NA) as substrate and digestion with Trypsin. Project File No. 5/4/1-25/2020-NCD-1. Sample processing protocol: For this, 1 ml of 1 mg/ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with Carbaryl (in Acetonitrile)(working concentration of 5 microMolar for 1 hour at 37 degree Celsius. Further, acetylation was done using 2NA (prepared in Acetonitrile) for 45 minutes at 37 degree Celsius. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 minutes followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree Celsius. These were then subjected to Trypsin digestion (2 microgram, Promega, MS grade).Next, samples to be digested by Trypsin were incubated for overnight at 37 degree Celsius. Trypsin digestion was stopped by adding 1 percent Trifluoroacetic acid solution. The sample clean-up with C18 resins column was performed and the aliquots were then sent for data acquisition. Data Processing protocol: The MaxQuant software (2.0.3.1) was used for the analysis of data. Under group-specific parameters Acetyl (Y), Acetyl K, Acetyl (N-term), and (Acetyl protein N-term) were selected as variable modifications whereas the Carbamidomethyl (C) was considered a fixed modification. The label-free quantification was done and the albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. For digestion, Trypsin enzyme was added to MaxQuant enzyme file. The albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. Description of the raw files is as follow. Carbaryl is Human serum albumin treated with Carbaryl and Carbaryl _2NA is Human serum Albumin treated with Carbaryl and then substrate (2NA) was added.

Project description:This is study done for academic purpose and funded by ICMR, New Delhi, India, sanctioned project titled- To study the acetylation of human serum albumin (HSA) in presence and absence of Aminocarb using 2-Naphthyl Acetate (2NA) as substrate and digestion with Trypsin. Project File No. 5/4/1-25/2020-NCD-1. Sample processing protocol: For this, 1 ml of 1 mg/ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with Aminocarb (in Acetonitrile)(working concentration of 5 microMolar for 1 hour at 37 degree Celsius. Further, acetylation was done using 2NA (prepared in Acetonitrile) for 45 minutes at 37 degree Celsius. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 minutes followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree Celsius. These were then subjected to Trypsin digestion (2 microgram, Promega, MS grade).Next, samples to be digested by Trypsin were incubated for overnight at 37 degree Celsius. Trypsin digestion was stopped by adding 1 percent Trifluoroacetic acid solution. The sample clean-up with C18 resins column was performed and the aliquots were then sent for data acquisition. Data Processing protocol: The MaxQuant software (2.0.3.1) was used for the analysis of data. Under group-specific parameters Acetyl (Y), Acetyl K, Acetyl (N-term), and (Acetyl protein N-term) were selected as variable modifications whereas the Carbamidomethyl (C) was considered a fixed modification. The label-free quantification was done and the albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. For digestion, Trypsin enzyme was added to MaxQuant enzyme file. The albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. Description of the raw files is as follow. Aminocarb is Human serum albumin treated with Aminocarb and Aminocarb _2NA is Human serum Albumin treated with Aminocarb and then substrate (2NA) was added.

Project description:This is study done for academic purpose and funded by ICMR, New Delhi, India, sanctioned project titled- To study the acetylation of human serum albumin (HSA) in presence and absence of Eserine sulphate using 2-Naphthyl Acetate (2NA) as substrate and digestion with Trypsin. Project File No. 5/4/1-25/2020-NCD-1. Sample processing protocol: For this, 1 ml of 1 mg/ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with Eserine sulphate (in Acetonitrile)(working concentration of 5 microMolar for 1 hour at 37 degree Celsius. Further, acetylation was done using 2NA (prepared in Acetonitrile) for 45 minutes at 37 degree Celsius. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 minutes followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree Celsius. These were then subjected to Trypsin digestion (2 microgram, Promega, MS grade).Next, samples to be digested by Trypsin were incubated for overnight at 37 degree Celsius. Trypsin digestion was stopped by adding 1 percent Trifluoroacetic acid solution. The sample clean-up with C18 resins column was performed and the aliquots were then sent for data acquisition. Data Processing protocol: The MaxQuant software (2.0.3.1) was used for the analysis of data. Under group-specific parameters Acetyl (Y), Acetyl K, Acetyl (N-term), and (Acetyl protein N-term) were selected as variable modifications whereas the Carbamidomethyl (C) was considered a fixed modification. The label-free quantification was done and the albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. For digestion, Trypsin enzyme was added to MaxQuant enzyme file. The albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. Description of the raw files is as follow. Eserine is Human serum albumin treated with Eserine sulphate and Eserine_2NA is Human serum Albumin treated with Eserine sulphate and then substrate (2NA) was added.

Project description:This is study done for academic purpose and funded by ICMR, New Delhi, India, sanctioned project titled- To study the acetylation of human serum albumin (HSA) in presence and absence of Rivastigmine using 2-Naphthyl Acetate (2NA) as substrate and digestion with Trypsin. Project File No. 5/4/1-25/2020-NCD-1. Sample processing protocol: For this, 1 ml of 1 mg/ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with Rivastigmine (in Acetonitrile)(working concentration of 5 microMolar for 1 hour at 37 degree Celsius. Further, acetylation was done using 2NA (prepared in Acetonitrile) for 45 minutes at 37 degree Celsius. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 minutes followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree Celsius. These were then subjected to Trypsin digestion (2 microgram, Promega, MS grade).Next, samples to be digested by Trypsin were incubated for overnight at 37 degree Celsius. Trypsin digestion was stopped by adding 1 percent Trifluoroacetic acid solution. The sample clean-up with C18 resins column was performed and the aliquots were then sent for data acquisition. Data Processing protocol: The MaxQuant software (2.0.3.1) was used for the analysis of data. Under group-specific parameters Acetyl (Y), Acetyl K, Acetyl (N-term), and (Acetyl protein N-term) were selected as variable modifications whereas the Carbamidomethyl (C) was considered a fixed modification. The label-free quantification was done and the albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. For digestion, Trypsin enzyme was added to MaxQuant enzyme file. The albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. Description of the raw files is as follow. Rivastigmine is Human serum albumin treated with Rivastigmine and Rivastigmine_2NA is Human serum Albumin treated with Rivastigmine and then substrate (2NA) was added.

Project description:This is study done for academic purpose and funded by ICMR, New Delhi, India, sanctioned project titled- To study the acetylation of human serum albumin (HSA) in presence and absence of Aldicarb using 2-Naphthyl Acetate (2NA) as substrate and digestion with Trypsin. Project File No. 5/4/1-25/2020-NCD-1. Sample processing protocol: For this, 1 ml of 1 mg/ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with Aldicarb (in Acetonitrile)(working concentration of 5 microMolar for 1 hour at 37 degree Celsius. Further, acetylation was done using 2NA (prepared in Acetonitrile) for 45 minutes at 37 degree Celsius. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 minutes followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree Celsius. These were then subjected to Trypsin digestion (2 microgram, Promega, MS grade).Next, samples to be digested by Trypsin were incubated for overnight at 37 degree Celsius. Trypsin digestion was stopped by adding 1 percent Trifluoroacetic acid solution. The sample clean-up with C18 resins column was performed and the aliquots were then sent for data acquisition. Data Processing protocol: The MaxQuant software (2.0.3.1) was used for the analysis of data. Under group-specific parameters Acetyl (Y), Acetyl K, Acetyl (N-term), and (Acetyl protein N-term) were selected as variable modifications whereas the Carbamidomethyl (C) was considered a fixed modification. The label-free quantification was done and the albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. For digestion, Trypsin enzyme was added to MaxQuant enzyme file. The albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. Description of the raw files is as follow. Aldicarb is Human serum albumin treated with Aldicarb and Aldicarb _2NA is Human serum Albumin treated with Aldicarb and then substrate (2NA) was added.

Project description:This is study done for academic purpose and funded by ICMR, New Delhi, India, sanctioned project titled- To study the acetylation of human serum albumin (HSA) in presence and absence of Glyphosate using 2-Naphthyl Acetate (2NA) as substrate and digestion with Trypsin. Project File No. 5/4/1-25/2020-NCD-1. Sample processing protocol: For this, 1 ml of 1 mg/ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with Glyphosate (in Acetonitrile)(working concentration of 5 microMolar for 1 hour at 37 degree Celsius. Further, acetylation was done using 2NA (prepared in Acetonitrile) for 45 minutes at 37 degree Celsius. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 minutes followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree Celsius. These were then subjected to Trypsin digestion (2 microgram, Promega, MS grade).Next, samples to be digested by Trypsin were incubated for overnight at 37 degree Celsius. Trypsin digestion was stopped by adding 1 percent Trifluoroacetic acid solution. The sample clean-up with C18 resins column was performed and the aliquots were then sent for data acquisition. Data Processing protocol: The MaxQuant software (2.0.3.1) was used for the analysis of data. Under group-specific parameters Acetyl (Y), Acetyl K, Acetyl (N-term), and (Acetyl protein N-term) were selected as variable modifications whereas the Carbamidomethyl (C) was considered a fixed modification. The label-free quantification was done and the albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. For digestion, Trypsin enzyme was added to MaxQuant enzyme file. The albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. Description of the raw files is as follow. Glyphosate is Human serum albumin treated with Glyphosate and Glyphosate _2NA is Human serum Albumin treated with Glyphosate and then substrate (2NA) was added.

Project description:This is study done for academic purpose and funded by ICMR, New Delhi, India, sanctioned project titled- To study the acetylation of human serum albumin (HSA) in presence and absence of Neostigmine methyl sulphate using 2-Naphthyl Acetate (2NA) as substrate and digestion with Trypsin. Project File No. 5/4/1-25/2020-NCD-1. Sample processing protocol: For this, 1 ml of 1 mg/ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with Neostigmine methyl sulphate (in Acetonitrile)(working concentration of 5 microMolar for 1 hour at 37 degree Celsius. Further, acetylation was done using 2NA (prepared in Acetonitrile) for 45 minutes at 37 degree Celsius. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 minutes followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree Celsius. These were then subjected to Trypsin digestion (2 microgram, Promega, MS grade).Next, samples to be digested by Trypsin were incubated for overnight at 37 degree Celsius. Trypsin digestion was stopped by adding 1 percent Trifluoroacetic acid solution. The sample clean-up with C18 resins column was performed and the aliquots were then sent for data acquisition. Data Processing protocol: The MaxQuant software (2.0.3.1) was used for the analysis of data. Under group-specific parameters Acetyl (Y), Acetyl K, Acetyl (N-term), and (Acetyl protein N-term) were selected as variable modifications whereas the Carbamidomethyl (C) was considered a fixed modification. The label-free quantification was done and the albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. For digestion, Trypsin enzyme was added to MaxQuant enzyme file. The albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. Description of the raw files is as follow. NMS is Human serum albumin treated with Neostigmine methyl sulphate and NMS_2NA is Human serum Albumin treated with Neostigmine methyl sulphate and then substrate (2NA) was added.

Project description:This is study done for academic purpose and funded by ICMR, New Delhi, India, sanctioned project titled- To study the acetylation of human serum albumin (HSA) in presence and absence of Paraoxon using 2-Naphthyl Acetate (2NA) as substrate and digestion with Trypsin. Project File No. 5/4/1-25/2020-NCD-1. Sample processing protocol: For this, 1 ml of 1 mg/ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with Paraoxon (in Acetonitrile)(working concentration of 5 microMolar for 1 hour at 37 degree Celsius. Further, acetylation was done using 2NA (prepared in Acetonitrile) for 45 minutes at 37 degree Celsius. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 minutes followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree Celsius. These were then subjected to Trypsin digestion (2 microgram, Promega, MS grade).Next, samples to be digested by Trypsin were incubated for overnight at 37 degree Celsius. Trypsin digestion was stopped by adding 1 percent Trifluoroacetic acid solution. The sample clean-up with C18 resins column was performed and the aliquots were then sent for data acquisition. Data Processing protocol: The MaxQuant software (2.0.3.1) was used for the analysis of data. Under group-specific parameters Acetyl (Y), Acetyl K, Acetyl (N-term), and (Acetyl protein N-term) were selected as variable modifications whereas the Carbamidomethyl (C) was considered a fixed modification. The label-free quantification was done and the albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. For digestion, Trypsin enzyme was added to MaxQuant enzyme file. The albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. Description of the raw files is as follow. Paraoxon is Human serum albumin treated with Paraoxon and Paraoxon _2NA is Human serum Albumin treated with Paraoxon and then substrate (2NA) was added.

Dataset Information

To study the acetylation of human serum albumin (HSA) in presence and absence of Chlorpyrifos-oxon using 2-Naphthyl Acetate (2NA) as substrate and digestion with Trypsin

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