How do you remove silver gel stains?
Remove the gel from the destain before complete destaining has occurred. Wash several times in deionized water to remove the destain and prevent fogging. Stop the destaining process with 10% acetic acid for 15 minutes. Wash the gel for at least 1 hour with several changes of water.
How does silver staining of proteins work?
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 stain reversible?
Coomassie staining is very frequently used with SDS-PAGE because it is reversible and is compatible with downstream mass spectrometric analyses. Silver staining relies on the reduction of bound silver ions, which result in distinct markings of metallic silver.
What is silver nitrate staining?
In traditional stained glass, silver stain is a technique to produce yellow to orange or brown shades (or green on a blue glass base), by adding a mixture containing silver compounds (notably silver nitrate), and firing lightly. It was introduced soon after 1800, and is the “stain” in the term “stained glass”.
Is silver staining toxic?
Current silver staining methods have had the disadvantages of long development time (12-24 hours), unstable solutions, and very toxic chemicals. Sigma’s method is 3.5 hours from fixed to stained gel. Sigma’s solutions are less toxic and stable as shipped.
Why is silver staining done?
Silver staining is a special yet powerful staining technique that is used for the detection and identification of proteins in gels. This is because silver binds to the chemical terminal or side chains of amino groups i.e carboxyl and sulfhydryl groups.
What is a silver stain used for?
Silver stain techniques are widely used to detect nanogram quantities of proteins following electrophoresis. Silver nitrate is the silver source in most silver stain procedures. Silver staining is a highly sensitive method for detecting proteins in polyacrylamide slab gels.
Why do we use silver stain?
Is silver staining more sensitive than Coomassie?
Coomassie blue staining is approximately 50-fold less sensitive than silver staining. However, due to its simplicity in binding, it is preferred in many studies.
What does silver stain stain?
Silver staining is an excellent technique for detecting proteins which are separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis due to its efficiency of detecting proteins present in nanograms.
What is silver stain used for?