Journal: International journal of oncology
Article Title: Proteomic identification of angiomotin by ProteomeLab PF-2D and correlation with clinical outcome in human clear cell renal cell carcinoma.
doi: 10.3892/ijo.2013.1889
Figure Lengend Snippet: Figure 1. Chromatographic analysis of proteins of RLC-310 and HK-2 cell lines using ProteomeLab PF 2D. (A) Chromatofocusing of total proteins in a pH gradient. In the chromatogram, the X-axis corresponds to the retention time in the column, the Y-axis (left) to the pH and the Y-axis (right) to the AU (UV absorbance). (B) Resolution of ProteomeLab PF-2D in a virtual gel given by ProteoVue software. Fifteen fractions were separated in pH gradient 8.5-4.5 and by hydrophobicity. Comparison of two aligned peaks in (A) displayed a quantitative difference in protein content measured by peak area (volume). Color scheme ranged from purple (low absorbance) to red (high absorbance). The difference between absorbencies was shown in the middle as either a red or green band, representing the sample with greater absorbance at a specific peak.
Article Snippet: The reagents used in the study are as follows: RPMI-1640 and fetal bovine serum (FBS) (Gibco/Invitrogen, Carlsbad, CA, USA); Transwell inserts (Costar, Cambridge, MA, USA); ProteomeLabTM PF-2D system and its matched separation kit (Beckman Coulter, Fullerton, CA, USA); LC-ESI-MS/ MS instrument (Thermo Finnigan US Companies, San Jose, CA, USA); rabbit anti-human Amot polyclonal antibody (Abcam, Cambridge, MA, USA); goat anti-rabbit secondary antibody and rabbit anti-human β-actin polyclonal antibody (Jackson Co., Lansing, MI, USA); TRIzol reagent (Invitrogen, Carlsbad, CA, USA); RNAfast200 (Flytech Biotechnology, Shanghai, China); one-step SYBR RNA PCR kit II (Takara Biomedical, Dalian, China); TPCK-trypsin, trifluoroacetic acid (TFA), n-octyl glucoside and protease inhibitors (Sigma Co., St. Louis, MO, USA).
Techniques: Software, Comparison
