I have been making hydrogels regularly using 10X DMEM (phenol red indicator), Ultrapure water, 1N NaOH, and 3.41 mg/ml Collagen (suspended with acetic acid). We have used the same calculations for a long time and within a two day period, the process started coming out around a pH of 4 when it is normally around 7.2-7.4. We have purchased all new components for making the hydrogel, initially thinking one of them was starting to degrade. Now, with everything new, we are still getting an acidic gel as the phenol red indicator in the 10X DMEM is going from red to yellow.
Steps:
1. 1.686% 1N NaOH.
2. 15% Ultrapure H2O.
3. 73.314% Collagen (bonded with acetic acid).
4. Thoroughly mix to bond acetic acid and NaOH. This should create a neutral solution.
5. 10% 10X DMEM.
We have tried this with cold, warm, and room temp components.
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