What is silver staining method?

What is silver staining method?

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.

How do you prepare silver stain?

Silver staining

1. Fix the gel in fixation solution (40% ethanol, 10% acetic acid, 50% water) for 30 minutes.
2. Treat the gel with protein treatment solution (20% ethanol, 5% acetic acid, 75% water, 4 mg dithiothreitol) for 30 minutes.
3. Rinse the gel with 0.5% dichromate for 5 minutes.
4. Wash the gel with water for 5 minutes.

Which can be used for staining protein?

Coomassie brilliant blue stain is easily available and is the most commonly used protein stain. However, staining and destaining require more time and reagents. The silver stain is the alternative colorimetric stain for increased detection sensitivity as compared to the Coomassie staining.

Which stain is used to stain protein bands in electrophoresis?

Coomassie Blue stain
Coomassie Blue stain is used to stain the protein bands in polyacrylamide gels. One common way to use it is to dissolve the dye in a mixture of methanol, acetic acid, and water. This stain will permeate the gel, stain the protein, and also fix the protein in place.

How does silver staining of proteins work?

Does silver stain stain DNA?

Currently, silver staining has widely been used to detect DNA fragments in various experiments, including short interspersed nuclear elements, variable number of tandem repeat detection and single nucleotide polymorphism study. The sensitivity of sil- ver staining is the same as that of radioisotopic methods.

Why is silver staining used?

In pathology, silver staining is the use of silver to selectively alter the appearance of a target in microscopy of histological sections; in temperature gradient gel electrophoresis; and in polyacrylamide gels.

What is the difference between silver stain and Coomassie?

Coomassie is much faster but 10-100XX less sensitive than silver stain. If your band is faint you should use silver, but its also a bit more tedious.

Is silver stain quantitative?

Silver staining has a narrow linear dynamic range (the range at which the level of staining is linear to the concentration), making it less suitable for quantification.

Which staining reagent is used for staining proteins?

The most common method of in-gel protein detection is staining with Coomassie dye. These stains either use the G-250 (“colloidal”) or the R-250 form of the dye. Colloidal Coomassie stains can be formulated to effectively stain proteins within 1 hour and requires only water (no methanol or acetic acid) for destaining.

Which type of detection is used for silver stain?

total protein
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.

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.