Journal: Clinical Proteomics

Article Title: A feasibility study to identify proteins in the residual Pap test fluid of women with normal cytology by mass spectrometry-based proteomics

doi: 10.1186/1559-0275-11-30

Figure Lengend Snippet: Protein content of residual Pap test fluid. (A) The protein concentration of residual Pap test fluid from 72 randomly selected SurePath TM samples was measured using the BCA protein assay in duplicate. Error bars show standard deviation. (B) The protein composition of the cell-free Pap test fluid from 16 individuals randomly selected from the samples in (A) was visualized by silver staining of SDS-PAGE gels. Five micrograms of protein was loaded per lane. MW, molecular weight standards. (C) Five micrograms of protein from serum (lane 6; S) or 5 different individuals’ SurePath TM samples (lanes 1–5) with varying protein concentrations (0.1–0.5 mg/ml) were visualized by silver stained SDS-PAGE. (D) In a parallel experiment, the SDS-PAGE gels was transferred to a PVDF membrane and probed by Western immunoblot with a polyclonal antibody recognizing human serum albumin. In (C) and (D) , the protein concentration of the individual’s Pap test sample is listed above each lane. The large arrow indicates the individual’s Pap test sample with the highest protein concentration (lane 2; 0.5 mg/ml), and the small arrow indicates the individual’s Pap test sample with the lowest serum albumin content as detected by Western immunoblotting (lane 4).

Article Snippet: One Pap test, which we used in this study, is the SurePath TM Pap test [Becton-Dickinson (BD Diagnostics, Burlington, NC)] which has an alcohol-based fixative consisting of 21.7% ethanol, 1.2% methanol, 1.1% isopropanol, and formaldehyde [ ].

Techniques: Protein Concentration, Bicinchoninic Acid Protein Assay, Standard Deviation, Silver Staining, SDS Page, Molecular Weight, Staining, Membrane, Western Blot

Journal: Clinical Proteomics

Article Title: A feasibility study to identify proteins in the residual Pap test fluid of women with normal cytology by mass spectrometry-based proteomics

doi: 10.1186/1559-0275-11-30

Figure Lengend Snippet: Diagrammatic representation of the workflow involved in Pap test sample preparation for MS analysis. Following a routine Pap test, the SurePath TM vials were sent to cytopathology for a diagnosis. Excess residual SurePath TM fluid from women with normal cytology was sent to the research laboratory. Protein concentration was determined by the BCA protein assay, proteins were precipitated with acetone, and visualized with silver stain by SDS-PAGE. Precipitated proteins were also trypsin digested and processed by FASP, and peptides were run on HPLC followed by MS. Data was analyzed by Sequest database searching and Scaffold analysis.

Article Snippet: One Pap test, which we used in this study, is the SurePath TM Pap test [Becton-Dickinson (BD Diagnostics, Burlington, NC)] which has an alcohol-based fixative consisting of 21.7% ethanol, 1.2% methanol, 1.1% isopropanol, and formaldehyde [ ].

Techniques: Sample Prep, Biomarker Discovery, Protein Concentration, Bicinchoninic Acid Protein Assay, Silver Staining, SDS Page

Journal: Clinical Proteomics

Article Title: A feasibility study to identify proteins in the residual Pap test fluid of women with normal cytology by mass spectrometry-based proteomics

doi: 10.1186/1559-0275-11-30

Figure Lengend Snippet: Protein content of residual Pap test fluid. (A) The protein concentration of residual Pap test fluid from 72 randomly selected SurePath TM samples was measured using the BCA protein assay in duplicate. Error bars show standard deviation. (B) The protein composition of the cell-free Pap test fluid from 16 individuals randomly selected from the samples in (A) was visualized by silver staining of SDS-PAGE gels. Five micrograms of protein was loaded per lane. MW, molecular weight standards. (C) Five micrograms of protein from serum (lane 6; S) or 5 different individuals’ SurePath TM samples (lanes 1–5) with varying protein concentrations (0.1–0.5 mg/ml) were visualized by silver stained SDS-PAGE. (D) In a parallel experiment, the SDS-PAGE gels was transferred to a PVDF membrane and probed by Western immunoblot with a polyclonal antibody recognizing human serum albumin. In (C) and (D) , the protein concentration of the individual’s Pap test sample is listed above each lane. The large arrow indicates the individual’s Pap test sample with the highest protein concentration (lane 2; 0.5 mg/ml), and the small arrow indicates the individual’s Pap test sample with the lowest serum albumin content as detected by Western immunoblotting (lane 4).

Article Snippet: The detachable head of the sampling device was immediately placed into a BD SurePath TM vial, which contains 10 ml of a mixture consisting of 21.7% ethanol, 1.2% methanol, 1.1% isopropanol, and formaldehyde [ ].

Techniques: Protein Concentration, Bicinchoninic Acid Protein Assay, Standard Deviation, Silver Staining, SDS Page, Molecular Weight, Staining, Western Blot

Journal: Clinical Proteomics

Article Title: A feasibility study to identify proteins in the residual Pap test fluid of women with normal cytology by mass spectrometry-based proteomics

doi: 10.1186/1559-0275-11-30

Figure Lengend Snippet: Diagrammatic representation of the workflow involved in Pap test sample preparation for MS analysis. Following a routine Pap test, the SurePath TM vials were sent to cytopathology for a diagnosis. Excess residual SurePath TM fluid from women with normal cytology was sent to the research laboratory. Protein concentration was determined by the BCA protein assay, proteins were precipitated with acetone, and visualized with silver stain by SDS-PAGE. Precipitated proteins were also trypsin digested and processed by FASP, and peptides were run on HPLC followed by MS. Data was analyzed by Sequest database searching and Scaffold analysis.

Article Snippet: The detachable head of the sampling device was immediately placed into a BD SurePath TM vial, which contains 10 ml of a mixture consisting of 21.7% ethanol, 1.2% methanol, 1.1% isopropanol, and formaldehyde [ ].

Techniques: Sample Prep, Protein Concentration, Bicinchoninic Acid Protein Assay, Silver Staining, SDS Page