Re: Antibody removal
Pete Macardle (Peter.Macardle@flinders.edu.au)
Wed, 27 Aug 1997 08:34:47 +0930 (CST)
>Date: Tue, 26 Aug 1997 13:45:28
>To: Kevin Holmes <KHOLMES@atlas.niaid.nih.gov>
>From: Pete Macardle <Peter.Macardle>
>Subject: Re: Antibody removal
>
>Hi Kevin,
>
>We have been trying to do this sort of thing, ie removing antibody from
stained cells but only one antibody at a time. We incubate the cells with an
anti-mouse antibody for an hour at room temp (22C) then wash the cells etc.
It works for some antibodies but not for all. It works , in our hands for CD3.
>
>The idea was based on the Dynal Detacha bead product. Which is also an
anti-mouse antibody directed to the Fab region of the monoclonal antibody.
Again their product works for some antibodies but not all. My understanding
of how it works is not terribly clear but I think it is supposedly due to
competitive inhibition of the binding site. Presumably the reason it works
for some and not all is due to the affinity of the MoAb.
>
>I'd be interested to hear of any other ideas that come up
>
>Best Wishes
>
>Pete
>
>At 04:40 PM 8/25/97 -0400, you wrote:
>>
>>An investigator in our Institute asked me a question that I'd like to
>>pass along:
>>He is doing a multicolor sorting experiment and wants to remove the
>>fluorochrome labeled antibody after sorting, before he puts the cells
>>into culture.
>>The only thing I could think of was trypsin treatment, but I was
>>unsure of the long term effects of this, or whether this would cause
>>more harm than good.
>>Any suggestions?
>>Thanks.
>>
>>Kevin L. Holmes, Ph.D.
>>Head, Flow cytometry Unit
>>Office of the Scientific Director
>>Bldg 7, Room 01
>>NIAID, NIH
>>
>>Phone: 301-496-9071
>>FAX: 301-402-4532
>>Email: kholmes@atlas.niaid.nih.gov
>>
>>
>>
>