Dear Morteta
0.2% v/v indeed means volume per volume (0.2 ml added to 100 ml in this case). In many other zymography protocols Brij35 is used in a concentration 0.02 % w/v. Commercialy available 10% Brij35 nead to be diluted as 0.2% v/v to yield an end concentration of 0.02% (w/v).
My guess would be that you dissolve 20 mg Brij35 powder to 100 ml of developing buffer. Be sure to mix it carefully (not to fast spinning) but thoroughly. Best to keep it on e.g. a roller bank for an overnight.
Good luck
Hello,
Check this link and try to procure something like this:
https://www.thermofisher.com/order/catalog/product/28316
It would be easier for you to prepare the working concentration of the developing buffer which should contain 0.02% Brij35, if you could have these kind of reagents in your lab. Usually, when you start to develop a new method it's better to look for the easiest solutions even if they are a little more expensive; in this way you are sure that you will have the reproducibility of the results, which is a must in order to go further with your research.
Good luck,
Mariana
Dear Duco Kramer & Mariana Constantinovici
Appreciate your kind and knowledge for both of you. Firstly, I'll try preparing 10% stock then will see the results!! if not good I'll try the commercial one with great hope to be the the right one coz they didn't mention the cata. no. of Brij35 in the paper.
Thank you again
Morteta
Dear Morteta H AL-Medhtiy
I would you like to Know whether it is a gelatin zymography? I regularly do gelatin zymography and i used to use Brij 35, but for the past 1 year i have omitted Brij 35 from the activity buffer and there is no change in the activity of MMPs. ou may try this once and for further details mail me at [email protected], I will send you detailed protocol.
Regards
Anil
Thank you so much Anil
I'm sending you an email and looking to hear from you soon please
Regards
Morteta
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