What does SYPRO Ruby stain?
SYPRO Ruby Protein Gel Stain is a highly sensitive, ready-to-use fluorescent stain for the detection of total proteins separated by polyacrylamide gel electrophoresis (PAGE). It is ideal for use in 1D and 2D PAGE. The sensitivity of SYPRO Ruby Gel Stain is as good as or better than the best silver staining techniques.
How does SYPRO Ruby stain work?
SYPRO Ruby protein gel stain is compatible with a variety of imaging platforms since it absorbs maximally in the ultraviolet (280 nm) and visible (470 nm) regions of the spectrum. Dye localization is achieved by noncovalent, electrostatic and hydrophobic binding to proteins, with signal being detected at 610 nm.
Can you reuse SYPRO Ruby?
Yes, we have determined that SYPRO Ruby Protein Gel Stain (1703126, 1703125 and 1703138) can be reused twice with no loss in sensitivity, if the gel is washed first, as described in our protocol. After two uses, there is an increasing loss of sensitivity, the more times it is used.
What is silver staining used for?
Silver staining is the most sensitive colorimetric method for detecting total protein. The technique involves the deposition of metallic silver onto the surface of a gel at the locations of protein bands. Silver ions (from silver nitrate in the staining reagent) interact and bind with certain protein functional groups.
Is silver staining more sensitive than Coomassie?
Coomassie Blue staining is approximately 50-fold less sensitive than silver staining, however due to its simplicity binding Coomassie Blue is preferred. Staining methods that allow destaining of gels makes them more appealing, as destaining may be needed for certain downstream applications.
How does Coomassie Brilliant Blue bind to proteins?
The Coomassie Brilliant Blue dye which is commonly used in SDS-PAGE, was first described by the German scientist Volker Neuhoff. In the staining reaction, the Coomassie dye binds to proteins through ionic interactions between sulfonic acid groups and positive protein amine groups through Van der Waals attractions.
Why does Coomassie blue change color?
When proteins bind to Coomassie blue in acid solution their positive charges suppress the protonation and a blue colour results. The binding of the dye to a protein causes a shift in the absorption maximum of the dye from 465 to 595 nm and it is the increase in absorbance at 595 nm that is monitored. Coomassie blue.
Does Coomassie stain all proteins?
As with all staining methods, Coomassie staining detects some proteins better than others, based on the chemistry of action and differences in protein composition. Thus, Coomassie staining can detect as little as 8–10 ng per band for some proteins and 25 ng per band for most proteins.
What is the most sensitive protein stain?
Silver staining
Silver staining is the most sensitive colorimetric method for detecting total protein. The technique involves the deposition of metallic silver onto the surface of a gel at the locations of protein bands.
What does warthin Starry stain?
Warthin – Starry stain is a stain based on silver nitrate used in histology. This type of stain is commonly called silver staining. Warthin – Starry stain is used for the visualization of spirochetes but also for the detection of Helicobacter pylori and microsperediates.
How is the Sypro Ruby protein blot stain used?
SYPRO Ruby Protein Blot Stain is a highly sensitive, ready-to-use fluorescent stain for proteins on nitrocellulose or PVDF membranes, making it easy to assess the efficiency of protein transfer to the blot and to determine if lanes are loaded equally.
What are the excitation maxima of Sypro Ruby?
Viewing and Photographing the Blot. SYPRO Ruby protein blot stain has two excitation maxima, one at ~280 nm and one at ~450 nm, and has an emission maxi- mum near 618 nm (Figure 1).
What makes syproruby protein gel stain an irritant?
SYPRO®Ruby protein gel stain is characterized as an irritant due entirely to the solvent system and buffer salts in the product. The heavy metal component of the stain (ruthenium) is not a regulated hazardous heavy metal in the United States.