I need to pre stain my protein samples before running sds page. Will mixing the coomassie brilliant blue help?
Remazol is the generic name of a group of dyes, widely used in the textile industry, which can covalently bind to proteins. The linkage of Remazol dyes to proteins is performed at alkaline conditions. The protein solution will be mixed with a 100mM NaCO3 solution, then the remazol is added to a final concentration of 2mg/ml and 2% SDS. This mixture should be incubated at 60 ºC for one hour. Then you can add sucrose or glycerol and load directly into the gel (or mixed with conventional loading buffer).
You can use the Remazol dye, however keep in mind that this can modify the relative mobility of pre-stained compared to unlabeled protein
Remazol is the generic name of a group of dyes, widely used in the textile industry, which can covalently bind to proteins. The linkage of Remazol dyes to proteins is performed at alkaline conditions. The protein solution will be mixed with a 100mM NaCO3 solution, then the remazol is added to a final concentration of 2mg/ml and 2% SDS. This mixture should be incubated at 60 ºC for one hour. Then you can add sucrose or glycerol and load directly into the gel (or mixed with conventional loading buffer).
I echo Dr Sepehrimanesh's answer... why bother. You can always stain with Coumassie afterwards. There are lots of kits with dyed proteins as molecular weight markers but as the others have pointed out you have to be careful since the derivitization can result in denaturation. None of this matters if you are using an SDS gel but with native acrylamide or agar gels it is a problem.
I typically use Ponceau-S Red to stain the proteins after transfer. This stain is reversible in tap water and does not affect the ability of primary antibody to react against target proteins. While, Ponceau-S works great on Nitrocellulose, Ponceau-S does not work well with nylon membranes. I attach a protocol for your use.
Remazol Blue is used to stain custom-prepared protein standards for PAGE; I've tried it myself before. Guillermo's point about changes in mobility is an important one to keep in mind when pre-staining.
Josh, just wanted to share that I've not had any trouble with Ponceau-S staining of PVDF membranes myself.
You can use Instant-Bands sample treatment buffer available from EZBiolabs for pre-staining of protein samples for SDS-PAGE. The sample buffer fluorescently stain protein samples, which can be visualised by UV transilluminator. Further details about the product available at the following web address
http://www.ezbiolab.com/instant-bands.html
Hi Souparno, Finally, did you try with coomasie? did it works? Because I have the same problem
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