RE: Making monoclonals to cell surface molecules

From: Moss, Delynn M. (dmm3@CDC.GOV)
Date: Thu Apr 26 2001 - 16:38:09 EST


Tony Schountz wrote:

		-----Original Message-----
		From:	Tony Schountz [mailto:tschount@mesastate.edu]
		Sent:	Wednesday, April 25, 2001 3:27 PM
		To:	Cytometry Mailing List
		Subject:	Making monoclonals to cell surface molecules


		I'd like to solicit opinions on producing monoclonal
antibodies to cell
		surface proteins. I will be immunizing BALB/c mice with
thymocytes or
		splenocytes from a deer mouse, a species that is much more
divergent
		from lab mice than rats are. I will be screening hybridoma
supes by flow
		cytometry so that any antibody I detect will be to cell
surface proteins
		only.

		One question that I have is whether it is necessary to
emulsify the deer
		mouse cells in CFA/IFA for immunization/boost. Most papers
are unclear
		on this, while some specifically say that this was done.
Goding's book
		states that this is unecessary, while Current Protocols in
Immunology
		doesn't really address whole-cell immunizations.

		Any tips or suggestions will be greatly appreciated.

		Thanks,

		Tony

		--
		Tony Schountz, Ph.D.
		Department of Biological Sciences
		Mesa State College
		mailto:tschount@mesastate.edu





		You might want to consider Hunter's TiterMax adjuvant.	It
is not as toxic as CFA and IFA, and I have gotten excellent results in
BALB/c that were immunized with protein.  There number is 800-345-2987.

		Good Luck

		Delynn Moss
		Centers for Disease Control



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