Journal: Physiological genomics

Article Title: Insights into a plasma membrane signature.

doi: 10.1152/physiolgenomics.2001.5.3.129

Figure Lengend Snippet: Fig. 1. Comparison of plasma mem- brane (PM) proteins by two-dimensional (2D) PAGE. PMs were isolated from four fibroblast (A–D; Hs 58.Fs, HE-SK, CCD-1090Sk, and Detroit 551) and four mammary carcinoma cell lines (E– H; MCF7, MDA-MB-231, MDA-MB-435, and SK-BR-3). Following membrane isolation, proteins were solubilized with a mixture of chaotropic and re- ducing agents and detergent as de- scribed in MATERIALS AND METHODS. A total of 100 mg of protein from this PM preparation was analyzed by 2D PAGE. Samples were focused in immo- bilized pH gradient (IPG) strips with a linear pH range of 4–7 and then were separated on a 12% acrylamide gel. Gels were silver stained and scanned, and the resulting image was analyzed using ImageMaster Analysis software. Only spots that constituted greater than 0.04% of the total staining den- sity within the gel were considered for analysis. All of the protein spots used for comparative analysis are shaded dark. Each gel is representative of at least three replicate gels.

Article Snippet: Solubilized PM protein, 100 mg in 200 ml of 8 M urea, 2 M thiourea, 2% CHAPS, and 13 mM DTT was used to rehydrate an 11-cm immobilized pH gradient (IPG) strip (Immobiline DryStrip, 4-7L; Amersham Pharmacia Biotech).

Techniques: Comparison, Clinical Proteomics, Isolation, Membrane, Acrylamide Gel Assay, Staining, Software