![]() Stain-Free gels have more reproducible data with smaller coefficients of variation compared to Coomassie or silver stains (McDonald et al. Some fluorescent stains can detect proteins at levels below the 1 or 0.5 ng limit. The limit of detection for the Stain-Free gels is 8 to 28 ng, similar to that of silver stains (0.6 to 1.2 ng), while Coomassie R-250 stain can detect protein amounts of at least 35 to 50 ng. The gel was imaged with a Stain-Free enabled imager, then stained with Coomassie (CBB R-250 and Bio-Safe G-250) stain and imaged on a densitometer. Serial 1:2 dilutions of broad range unstained molecular weight standards were separated on a 4–20% Criterion Stain Free Tris-HCI Gel. Comparison of a Stain-Free gel, CBB R-250, and Bio-Safe G-250 stained gel images. For proteins with higher tryptophan content, Stain-Free gels provide much higher sensitivity than CBB-stained gels (Figure 1).įigure 1. In general, the sensitivity of Stain-Free gels when visualizing data is equal to that of Coomassie-stained gels for all proteins. Protein visualization data obtained from Stain-Free gels are comparable to those obtained from gels stained with other dyes. Stain-Free Technology Provides More Sensitivity and Better Dynamic Range than Coomassie Stains Stain-Free imaging allows for the elimination of the inherently problematic use of housekeeping proteins as loading controls on western blots, enabling the user to obtain truly quantitative western blot data by normalizing bands to total protein in each lane.Īdvantages of Using Stain-Free Technology ![]() With the fluorophore covalently bound to the protein molecules, they can be imaged repeatedly on a gel or membrane after transfer, without additional staining and destaining steps. The addition of the fluorophore allows visualization of proteins in the gel and following transfer onto a membrane during western blotting but does not interfere with electrophoresis or downstream steps. This trihalo compound is covalently bound to tryptophan residues, enhancing their fluorescence when exposed to UV light, enabling the detection of proteins at levels as low as 10–25 ng. ![]() Stain-Free imaging technology utilizes a polyacrylamide gel containing a proprietary trihalo compound to make proteins fluorescent directly in the gel with a short photoactivation, allowing the immediate visualization of proteins at any point during electrophoresis and western blotting. ![]()
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