Journal: BMC Cancer

Article Title: CUB Domain Containing Protein 1 (CDCP1) modulates adhesion and motility in colon cancer cells

doi: 10.1186/1471-2407-14-754

Figure Lengend Snippet: Analysis of CDCP1 and CD9 in detergent-resistant membrane (DRM) fractions of HT-29 colon cancer cells. HT-29 cells were plated at 3.8 × 10 6 cells per T175 flask and grown to confluence. The cells were scraped into MBS containing either 0.5% Triton-X100, Brij58 or Brij97 on ice. The lysates were fractionated by sucrose density gradient centrifugation as described in Materials and Methods. Eleven 0.5 ml fractions were collected, with fraction 1 being the top and fraction 11 the bottom of the gradient. The fractions were analysed by Western blotting using the following antibodies (final concentrations in parentheses): CDCP1 (1 μg/ml goat ab1377), CD9 (2 μg/ml mouse monoclonal 602–29), TfR (100 ng/ml mouse monoclonal H68.4 (13–6800)), Flotillin-1 (50 ng/ml). A, B: Triton X-100 extraction; C, D: Brij58 extraction; E, F: Brij97 extraction. B, D, F: Quantitation of the Western blots shown in panels A,C and E respectively using Advanced Image Data Analyzer (AIDA) software.

Article Snippet: Images were captured using a chemiluminescence imager (Fuji LAS-3000, Japan) and analysed using AIDA software (Raytek Scientific Ltd, Sheffield, UK).

Techniques: Membrane, Gradient Centrifugation, Western Blot, Extraction, Quantitation Assay, Software

Journal: BMC Cancer

Article Title: CUB Domain Containing Protein 1 (CDCP1) modulates adhesion and motility in colon cancer cells

doi: 10.1186/1471-2407-14-754

Figure Lengend Snippet: Co-immunoprecipitation of CDCP1 and CD9. SW480 cells were lysed in 1% (w/v) Brij97, 10 mM Tris–HCl (pH 7.4), 150 mM NaCl, 1 mM CaCl 2 , 1 mM MgCl 2 . A. Immunoprecipitation from whole cell extracts of SW480 cells was performed using 5 μg of ALB6 anti-CD9 monoclonal antibody (lane 6) or isotype-matched control antibody (lane 5). Immunoprecipitates were analysed by Western blotting with antibodies against either CDCP1 (ab1377), top panel; EpCAM (ab32392), middle panel or CD9 (602.29), lower panel. Further controls for the specificity of immunoprecipitation and Western blotting are: lane 1: 15 μg whole cell extract, lanes 2 and 3: 5 μg of isotype-control antibody and anti-CD9 antibody respectively; lane 4: SW480 proteins bound to protein G beads. B. Densitometric analysis of the Western blot for CDCP1. A portion of the blot corresponding to the CDCP1 p135 region was analysed in the tracks generated by immunoprecipitation with the control (red) and anti-CD9 (green) antibodies using AIDA software.

Article Snippet: Images were captured using a chemiluminescence imager (Fuji LAS-3000, Japan) and analysed using AIDA software (Raytek Scientific Ltd, Sheffield, UK).

Techniques: Immunoprecipitation, Control, Western Blot, Generated, Software

Journal: Retrovirology

Article Title: Highly conserved serine residue 40 in HIV-1 p6 regulates capsid processing and virus core assembly

doi: 10.1186/1742-4690-8-11

Figure Lengend Snippet: Mutation of Ser-40 has no effect on ALIX mediated virus release . (A) 293T cells were cotransfected with HIV-1 ΔPTAP or HIV-1 ΔPTAP/S40F and increasing amount of ALIX plasmid. (B) Quantification of Western blot data using AIDA Software (Raytest). Shown is the amount of Gag release determined by the ratio of virus associated Gag/total Gag expressed. (C) The amount of infectious units released from the cells was analyzed by β-galactosidase quantification after infection of TZM-bl cells.

Article Snippet: Radioactivity in dried gels was quantified using AIDA imaging software (Raytest).

Techniques: Mutagenesis, Virus, Plasmid Preparation, Western Blot, Software, Infection