class switching M hybridomas

[Richard R. Hardy]

In article <MAPI.Id.0016.0075726e732020203341464630303033 at MAPI.to.RFC822>,
burns at sasa.gov.uk (Robert Burns) wrote:

>Hi all,
>       does anybody out there have experience of class switching M
>hybridomas to G. I don't have access to a FACS for sorting, so it
>would have to be done manually by cloning. I have heard of people
>doing it by this method and wondered how feasible it really was.
>Also I know that highly specific isotyping reagents would be needed
>to identify the class-switched clones. Many of the reagents we have
>looked at across the years give some cross reactivity with some
>other isotypes, does anybody know of a good source?
>
I've done this by FACS, not sure how well it would work by limit diln (our
estimates of frequency varied from 1/10^6 to 1/10^5).  As to source of
anti-isotype reagents, you could try PharMingen (San Diego) - I think they
have monoclonal anti-isotype reagents.  Good luck!
____________________________________________________________________________
Richard R. Hardy                       Member, Institute for Cancer Research           
Fox Chase Cancer Center                Tel:    (215) 728-2463
7701 Burholme Ave.                     FAX:    (215) 728-2412
Philadelphia, PA 19111                 E-MAIL: R_HARDY at fccc.edu