Simply Blue SafeStain is a ready-to-use Coomassie G-250 stain that does not require hazardous disposal. It is made with no methanol or acetic acid and, therefore, can be safely dumped into the sink. The storage is at room temperature and the product is guaranteed to be stable for at least 6 months. The detection of proteins is fast and sensitive. Simply Blue SafeStain comes in two standard sizes: 1 and 3.5 L. The standard 1 L volume provides sufficient reagent to stain 50 mini-gels. Alternatively, 3.5 L bottle comes with a pump dispenser and is more economically priced for the laboratories with a high volume of protein experiments.
Two different protocols are available for general staining with Simply Blue SafeStain. The basic protocol is generally used with some modifications to achieve maximum sensitivity. First, the gel is washed and rinsed three times in water and then placed in the Simply Blue SafeStain for at least 1 h (with gentle agitation). After staining, the gel is washed one or two times for 1-3 h in water. For increased sensitivity, the slightly modified procedure with addition of salt can be employed. After staining and washing the gel, 20% NaCl is added to the second wash (can be done overnight). The other fast procedure is called a microwave protocol. The basic steps of washing, staining and washing are the same, but the length of all the steps is greatly reduced from hours to minutes. The whole process takes about an hour, unless an extra clear background is required, then an extra hour long wash in the water is needed.
For the last several months, I have routinely used the microwave protocol for staining 1.0 mm NuPage Novex Bis-Tris gels, manufactured by Invitrogen. The protocol consists of steps that are easy to perform and utilizes equipment available in every research laboratory. I found the results of the Simply Blue SafeStain to be very robust and reliable. Routinely, brightly stained blue bands appeared on my gels almost immediately after the start of incubation. The stain itself is very safe and easy to work with, much better than previous methods. I found that the Simply Blue SafeStain protocol gave equivalent staining to previous methods, but the overall protocol was faster since I didn’t have to fix or de-stain.
I cannot comment on the sensitivity of Simply Blue SafeStain. The proteins I analyzed were made in large quantities through the in-vitro transcription-translation system. Therefore, I had no opportunity to test how sensitive the stain actually is. To achieve a clear background I routinely included an extra washing step into my procedure.
Because of the dramatic time difference between performing basic and microwave protocol, I have chosen the latter and noticed that there is a potential danger in the transfer of heated liquids between a microwave and shaker. You need to perform the procedure carefully and slowly. In summary, I am very satisfied with the reliable and consistent results obtained with Simply Blue SafeStain.
Yelena Bykhovskaya PhD
Research Scientist I
Molecular Hematology Laboratory
Cedars-Sinai Medical Center Los Angeles CA