Hybridoma Cell lines - How to do it (Oct/09/2005 )

We just got 2 new hybridoma cell lines. Thing is I'm not sure how to take care of them as I'm rather new to this? Do I have to culture them in low IgG Serum or anything special? What is a good Media recipe?

Does anyone have protocols/hints for Protein G column purification?


Thanks,

Moutainman

---

Whomever/wherever you got them from should also tell you what they were growing in before so they are not shocked, but it should be some rather standard media if they are stable lines.
(ie. DMEM w/ L-glutamine, 10% FBS, and bets-mercaptoethanol. There could also be some HCF (Hybridoma Cloning Factor) but hopefully they are weened off that expensive stuff already)
Looking on ATCC will give you media formulations for some fusion clones they have.

Low Ig serum will certainly keep the bovine Ig down for purification as would converting to serum free but that's finicky. It may depend on how much Ab you need and how pure it has to be.

G protein purification is very standard, a Google will get you some protocols, or looking at protocols on manufacturers of kits if you do not want to buy a kit will work.

---