b'Staining gels Protein stainsOnce protein bands have been separatedFluorescent and specialty stainsby electrophoresis, they can be directly Invitrogen SYPRO Orange, SYPRO Red, SYPRO Ruby gel stainsvisualized using different methods of in-gelThermo ScientificPierceReversible Protein Stain Kit for detection. Over the past several decades,Nitrocellulose or PVDF Membranesdemand for improved sensitivity and Invitrogen Pro-Q Emerald glycoprotein staincompatibility with downstream applications Invitrogen Pro-Q Diamond phosphoprotein stainand detection instrumentation has driven To visualize the proteins, a protein-specific, dye-binding the development of several basic stainingor color-producing chemical reaction must be performed methods. Each method has particularon the proteins within the gel. Depending on the particular chemistry of the stain, various steps are necessary to hold advantages and disadvantages, and athe proteins in the matrix and to facilitate the necessary number of specific formulations for each typechemical reaction. Most staining methods involve some version of the same general incubation steps:of method provide optimal performance for2 F DI H 2 O SDI H Ovarious situations.1. Water wash 2. Fix3. Water wash 4. Stain 5. Destain A water wash to remove electrophoresis buffers from the Typically, these stains can be classified broadly based ongel matrixthe molecules that help visualize the proteins: An acid or alcohol wash to condition or fix the gel to limit Coomassie stains diffusion of protein bands from the matrix Thermo Scientific PageBlue Protein Staining Solution Treatment with the stain reagent to allow the reagent toInvitrogen SimplyBlue SafeStain diffuse into the gel and bind to (or react with) the proteins Thermo Scientific Imperial Protein StainDestaining to remove excess dye from the background gel matrixSilver stains Thermo Scientific Pierce Silver Stain Kit Depending on the particular staining method, two or moreof these functions can be accomplished with one step.Invitrogen SilverXpress Silver Staining Kit For example, a dye reagent that is formulated in an acidicThermo Scientific Pierce Silver Stain forbuffer can effectively fix and stain in one step. Conversely, Mass Spectrometry certain functions require several steps. For example, silver staining requires both a staining reagent step and a development step to produce the colored reaction product.Learn more at thermofisher.com/proteinstains63'