| Jul. 9th, 2008 @ 06:04 pm (no subject) |
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Has anyone ever made collagen gels (or something similar), and if so, do you have any tips?
My collagen refuses to polymerize. I've done everything. I've incubated in 37°C with and without 5% CO2, room temp, etc. I've put it on slides, cover slips, treated plates, untreated plates, deep 24-well plates, thinner p100s. I've tried switching media. I've tried using NaOH vs. sodium bicarbonate to adjust the pH. I've tried switching the collagen from type I to II or III, and I've tried it with old collagen, new collagen, red collagen, blue collagen. I've tested the pH at every single step of the process. And it's not like it's a hard process, either - thaw collagen, add media, adjust pH with NaOH to taste, watch collagen gel, rejoice.
The closest I ever got was when I accidentally left a slide uncovered, and it dried out. The collagen either precipitates into a goopy mess, or it begins forming a matrix but never actually solidifies to a gel. And perhaps this is where I'm going wrong - is it even possible to get collagen to gel to a point where you can cut it into bits? I'm talking maybe jello consistency. (Preferably lime jello, but I'm willing to accept raspberry, as well.)
If this doesn't work I'm going to have to start sacrificing chickens and/or virgins. |
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