DIGE简介.pptVIP

DIGE简介 Advantages: Sample preparation and labelling Labelling reaction 2DExperimental design For example * 辉骏生物：/ 免费服务热线：400-699-1663 Description of Ettan DIGE system 辉骏生物：/ 免费服务热线：400-699-1663 1: Accurate quantification and accurate spot statistics between gels; 2: Increased confidence in matching between gels（ internal standard）; 3: Flexibility of statistical analysis depending on the relationship between samples; 4: Separation of induced biological change from system variation and inherent biological variation. 5. The dyes afford great sensitivity with detection down to 125 pg of a single protein, and a linear response to protein concentration up to five orders of magnitude (105). In comparison, silver stain detects 1–60 ng of protein with a dynamic range of less than two orders of magnitude. 辉骏生物：/ 免费服务热线：400-699-1663 ? Ensure the sample is prepared in a buffer that is compatible with the labelling method.? Ensure the sample protein concentration is 5-10 mg/ml.? Ensure the sample pH lies in the range pH 8.0–9.0.? Create a pooled internal standard from all samples for inclusion on every gel.? For labelling, an aliquot of the CyDye DIGE Fluor minimal dye stock solution should be diluted to a concentration of 400 pmol/μl.? The ratio of protein to CyDye DIGE Fluor minimal dye should be maintained at 50 μg: 400pmol. 辉骏生物：/ 免费服务热线：400-699-1663 There are three CyDye DIGE Fluor minimal dyes available; Cy2, Cy3 and Cy5. These have been designed to be matched for charge and molecular weight. The lysine amino acid in proteins carries an intrinsic +1 charge at neutral or acidic pH. CyDye DIGE Fluor minimal dyes also carry a +1 charge which, when coupled to the lysine, replaces the lysine’s +1 charge with its own, ensuring that the pI of the protein does not significantly alter. Consequently the same protein labelled with any of the CyDye DIGE Fluor minimal dyes, will migr