3D Collagen Matrix Polymerization - Technical Issue (Sep/15/2009 )
I'm trying to get a 3D collagen matrix but I cannot managed to got polymerization.
This is my protocole for 2,5ml of solution:
- 250無 of 10X PBS medium.
- 250無 of 10X Phenol Red solution.
- 625無 of collagen I solution at 1mg/ml. My stock solution of collagene is from rat tail (Sigma Lab).
- 1375無 of steril water.
- 10無 of NaOH to adjust pH.
I prepared my solution on ice and then put it on bacterial dishes. I wait for polymerization at 37蚓 (98,6蚌) under 5% CO2. The gel polymerization never happen!
I hope someone had an idea to help.
Thank you everyone.
Sorry it's not an answer but I had same problem with BD collagen I. I tried heating and adding acid both didn't work. I gave up and use matrigel now. You may refer to Bissell lab protocol at http://www.ihcworld.com/_protocols/lab_pro...b-protocols.htm.