The kit contains prepared solutions of silver staining reagents along with detailed instructions to achieve optimal results. Silver staining is used to detect proteins after electrophoretic separation on polyacrylamide gels. Silver staining techniques require containers to be thoroughly cleaned before use. Begin at step 2 of the silver staining protocol and proceed as usual. Switzer iii neuroscience associates, knoxville, tennessee 37922 abstract silver staining procedures have been used in numerous ways to render a variety of physical and biological features visible. This video explains the silver staining process and its applications in structural biological studies. Completely destain the coomassie stained gel in 40% methanol10% acetic acid.
This protocol is optimized for the following precast gels from. Readytouse solutions and preweighed chemicals minimise preparation time and maximize ease of use and reliability. The silver stain protocol has a flexible gel fixation step. Grocottgomoris methenamine silver gms stain is a histological stain that is used majorly for the identification of carbohydrates in fungal microorganisms. Silver staining of proteins in polyacrylamide gels mireille. Comparison of identical 2d gels stained with silver stain kit and another popular commercial silver stain formulation. The stain performs consistently and reliably for both firsttime and experienced users for high, low or gradient percentage gels in singledimension or 2d format.
Fast silver staining protocol used by the price lab. Protocol for silver staining of proteins scientist solutions. Staining containers may contain residues from soap or prior staining solutions, which silver will bind with to form metallic silver. After washing containers with laboratory soap and water, rinse with 50% nitric. Many silver staining protocols and commercial staining kits are not compatible with mass spectrometry. This protocol is valid for both sdspage and native page gels. Staining proteins in gels sasse 2009 current protocols.
Application of silver degeneration stains for neurotoxicity testing robert c. In this tutorial, you will learn the best protocol for staining your gels using proteas silver staining kit. Silverxpress silver staining protocols developer 5 ml ultra pure water 95 ml final volume. Silver staining of proteins in polyacrylamide gels. Mar 22, 2018 advances in impregnation procedures reduced time required for silver staining usually accomplished by rapidly increasing the temperatures of solutions using microwave techniques. Hiper protein silver staining teaching kit p r o d u c t i n f o r m a t i o n the information contained herein is believed to be accurate and complete. This protocol is compatible with maldimsms analysis of protein bands.
Silver staining of proteins in polyacrylamide gels nature. It combines excellent sensitivity in the low nanogram range whilst using very simple and cheap equipment and chemicals. Fungi, basement membranes, and reticulum can be stained with procedures in which oxidation of carbohydrates and glycoproteins is followed by reduction of silver by the oxidized molecule. Jul 15, 2012 the gallyas silver stain is a classic neuropathology method used for the detection of neurofibrillary tangles. It is used to show both substances inside and outside cells. Theory and practice of silver staining in histopathology. An example of a gel stained with the silverquest silver staining kit using the basic staining protocol is shown below. Pdf development of a simple and effective silver staining protocol. Pdf this protocol describes a simple silver staining method used to visualize dna fragments and other organic molecules with unsurpassed. It is compatible with downstream processing such as mass spectrometry analysis after protein digestion. It can be used to stain axons, neurofibrils and senile plaques in the central nervous system. The general principles underlying these silver staining techniques and their effective use are described in this manusc. However no warranty or guarantee whatsoever is made or is to be implied with respect to such information or with respect to any product, method or apparatus referred to herein.
The silver precipitates on the staining containers. Silver staining is used for sensitive detection of proteins separated by 1d and 2d sds page with detection limits from 0. Prepare destain by combining equal parts of solutions a and b just prior to use. The proteosilver silver stain kit is an exceptionally sensitive protein detection silver stain kit.
It is suitable for staining both single dimension sdspage gels and twodimensional 2d gels of complex protein solutions, a common characterization technique used in proteomics. Shalini jain, hariom yadav and pr sinha animal biochemistry division, national dairy research insitute, karnal2001, haryana, india. The pierce silver stain kit provides consistency, high sensitivity, low background and a flexible protocol. Silver staining kit, protein is a reagent kit for the fast, easy, reproducible and sensitive staining of proteins in polyacrylamide gels. Pdf silver staining dna in polyacrylamide gels researchgate. Application of silver degeneration stains for neurotoxicity.
Double and single stranded dna and rna in polyacrylamide gels can be stained. Cut silver stained bands from the gel and put in individual microfuge tubes. The fast staining protocol using a microwave oven for silver staining novex gels using silverquest silver staining kit is described below. This method is easy to perform and is routinely used in the. Compared with traditional silver staining, a significantly less amount of silver nitrate is required for this new staining techniquespecifically, 0. Future trends for silver staining will also be discussed. Its mission is to make life science research more efficient and reproducible by curating and hosting high quality, free access protocols.
Many silver staining protocols and commercial staining kits are not compatible with mass spectrometry due to the use of crosslinking reagents. Pdf silver staining is one of the widely used methods for dna fragment detection in biological research. Gel compatibility with the thermo scientific pierce silver stain kit. Nov 22, 2006 silver staining is used to detect proteins after electrophoretic separation on polyacrylamide gels. Fast silver staining protocol used by the price lab source. In all solutions containing formaldehyde fm, add the fm just before adding the solution to the gel. Avoid the use of metal containing materials in this procedure.
It combines excellent sensitivity in the low nanogram range with the use of very simple and. Decreased concentration of the silver solutions used, reduced costs and background staining. This protocol describes a silver nitrate staining approach. Original version original version updated version updated. Silver staining of protein gels md anderson cancer center.
Fluorescent silver staining of proteins in polyacrylamide. Using the silver staining protocol described on previous pages, you should be able to detect. In order to obtain a positive stain, one must use the complexing power of proteins for silver ion. Protocol for silver staining of gels optimized for mass spectrometry and protein identification guidelines silver staining is used for sensitive detection of proteins separated by 1d and 2d sds page with detection limits from 0. To summarize these data, we can consider that every staining protocol is divided into five main. The silverquest silver staining kit or silverxpress silver staining kit page 25 is suitable for silver staining of nativepage gels. Its sensitivity is in the lownanogram range, which is 50100 times more sensitive than classical coomassie blue staining, 10 times better than colloidal coomassie blue staining, and at least twice as sensitive as the zincimidazole negative staining method. The nativepage novex bistris gels are compatible with any of the standard coomassie staining. Silver staining is not recommended for agarose gels. Before beginning, make sure that all glassware is acid washed. Silver staining protocol compatible with mass spectrometry modified from mass spectrometric sequencing of proteins from silverstained polyacrylamide gels a. Fluorescent staining is a popular alternative to traditional staining procedures, mainly because it is more sensitive than coomassie staining, and often as sensitive as silver staining.
For the detection of proteins, a number of techniques have been developed in which silver staining is one of the most sensitive techniques available for. Careful manufacturing procedures and thorough quality testing ensure that the kit yields excellent results with every use, independent of the polyacrylamide gel type or buffer system being used. If not better, these homemade solutions work as good as commercially available ones. Silver staining is also used in temperature gradient gel electrophoresis. Proteosilver silver stain kit silver staining kit sigma. Proteosilver utilizes silver nitrate, which binds to. The red book bulletin, supplement 11 soaks and washes 250 ml are carried out in a clean glass tray scrubbed with 409. Alternate protocols describe rapid coomassie and silver staining methods, as well as fluorescent stains that are specific for phosphoproteins and glycoproteins.
Silver staining, widely regarded as the standard by which all other ultrasensitive staining methods are judged, is the most complex and variable protein gel staining methodology, with many dozens of published protocols, all of them requiring several stepwise procedures. Silver nitrate stains the nucleolar organization region norassociated protein, producing a dark region wherein the silver is deposited and denoting the activity of rrna genes within the nor. Mar 23, 2011 in this tutorial, you will learn the best protocol for staining your gels using proteas silver staining kit. Check what is needed by reading the procedure first.
It combines excellent sensitivity in the low nanogram range with the use of. Bielschowskys silver stain protocol by paul polak and douglas feinstein, university of illinois, chicago. For best results and better background, we recommend using the silverquest silver staining kit. All silver staining protocols, starting form the pioneer work 2 are made of the same basic steps, which are. This staining method was named after gyorgy gomori, a physician from hungary, who developed the staining methodology. Bielschowskys silver stain a very useful tool to detect nerve fibers. It can be hypothesized that the success of the new stain is purely down to the combination of the incredibly bright. Published september 28, 2012 staining proteins following sdspage or 2dimensional electrophoresis is a very useful technique for visualising a global population of proteins or checking expression of your recombinant proteins, but how do you know which stain to use.
Silver staining is the use of silver to stain histologic sections. Human chromosomes, 14, 15, 21, and 22 have nors, which increase the silver stain activity by at least 50 times. Silver staining protocol mass spectrometry facility. The silver stain protocol has staining flexibility. For these reasons, protocols based on silver nitrate are of a more general use and might be favored. Silver staining of proteins in polyacrylamide gels ncbi. Silver staining is used for sensitive detection of proteins separated by 1d and 2d sds page with detection limits from. Conveniently packaged, extremely stable, and highly sensitive, proteosilver is an ideal product for any protein scientist. This kind of staining is important especially to show proteins for example type iii collagen and dna. For the basic protocol and more details on the staining procedure, refer to the silverquest silver staining kit manual im6070.