As requested by some of you, here is our recipe for Intraprep reagents: Reagent 1: We make a 10% stock of para-formaldehyde in PBS, then dilute to 5.5% (v/v) with HBSS. Reagent 2: We make a 10% stock of saponin in HBSS, then make a solution of 0.1% to 0.2% saponin in HBSS and add 0.1% Na azide. I am sure you could use PBS also for both reagents. Our saponin comes from Sigma, Cat.#S-2149. You can also try the more pure saponin, Cat.# S-4521. In another lab, I found that the source and preparation of saponin can give different permeabilization results. Depending on your cells, you may have to adjust the concentration of saponin to get a good stain, but we normally use 0.1% saponin solution for many antibodies. We also added a wash between Reagent 2 and the addition of our antibodies. Other than that, just follow the same procedure they give you in the data sheet. Good luck. Dr. Susan J. Zunino Lehrstuhl fur Genetik University of Erlangen Staudtstr. 5 91058 Erlangen Germany Tele. 49 9131 8528784 FAX 49 9131 8528526 e-mail: szunino@biologie.uni-erlangen.de
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