3 replies to this topic

[anonymous]

I need a protocol for charcoal-stripping of serum.

[anonymous]

This works for me!

1. Make a 1.0M stock of Tris Buffer (ph 8.0, 4C).2. Combine 2.5g washed, activated charcoal, 0.25g dextran (T-70), q.s to 1.0L with 0.01M Tris (ph 8.0, 4C).3. Stir overnight at 4C.4. Autoclave the next day.5. Add an equal volume of dextran coated charcoal suspension to serum. 6. Stir at 45C for 1h.7. Centrifuge aliquots to remove charcoal (3000rpm, 5min). Often this requires multiple spins.8. Dectant the 50% serum into sterile containers.

Lately, I've just been buying the CS-FBS from Hyclone. It works great!

Good luck!

[anonymous]

This works for me!

1. Make a 1.0M stock of Tris Buffer (ph 8.0, 4C).2. Combine 2.5g washed, activated charcoal, 0.25g dextran (T-70), q.s to 1.0L with 0.01M Tris (ph 8.0, 4C).3. Stir overnight at 4C.4. Autoclave the next day.5. Add an equal volume of dextran coated charcoal suspension to serum. 6. Stir at 45C for 1h.7. Centrifuge aliquots to remove charcoal (3000rpm, 5min). Often this requires multiple spins.8. Dectant the 50% serum into sterile containers.

Lately, I've just been buying the CS-FBS from Hyclone. It works great!

Good luck!

[Sean Peel]

If you go to www.hyclone.com (who sell FBS) under their support section they have 2 places to look:

1) FAQ section reguarding serum components - they say they are willing to provide average values for a variety of components. just e-mail them

2) Art to Science - this is a very informative newsletter that discusses all sorts of things related to cell culture.