Trypsin and Annexin

Dr. Szollosi Janos szollo at jaguar.dote.hu
Tue Aug 25 02:29:18 EST 1998


In our hand trypsin caused only a 5-10 % decrease in the expression level
of erbB2 protein, suggesting that not all cell surface proteins are
stripped of during trypsin treatment. 

We compared trypsinized cells with EGTA harvested ones.  

I think that the stripping effect depends upon the cell lines and the
receptors you investigate. 


Regards

Janos


At 03:11 PM 8/19/1998 -0500, you wrote:
>
>Well, Trypsin has the reputation of stripping cell surface markers, and
>can't it kill off cells if the concentration is strong enough? Also, if
>cells surface markers are stripped and cells are starting to die, or are
>damaged somehow, they can exert autoflourescense.....or so I have found
>with cultured cells. Sorry, no exp. with Annexin, but I will soon as my
>kit arrived yesterday! 
>
><+>:<+>:<+>:<+>:<+>:<+>:<+>:<+>:<+>:<+>:<+>:<+>
>
>Patricia L. Echeagaray;  echeagaray at sri.org
>Manager of Flow Cytometry Services
>Southern Research Institute - Frederick
>Pharmaceutical Research and Development  Dept.
>431 Aviation Way     
>Frederick, Maryland 21701
>USA
>Voice: (301) 228-2170 or (301) 694-3232 ext 111 or 276
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>    echeagaray at sri.org
><+>:<+>:<+>:<+>:<+>:<+>:<+>:<+>:<+>:<+>:<+>:<+>
>
>
>>----------
>>From: 	Joan Kalnitsky[SMTP:jkalnits at mail.vt.edu]
>>Sent: 	Tuesday, August 18, 1998 10:11 AM
>>To: 	Cytometry Mailing List
>>Subject: 	Trypsin and Annexin
>>
>>
>>I have a client who is staining with Annexin-FITC.  They are looking at
>>cell lines and are using trypsin to get the cells off the flasks.  We are
>>seeing what we think is an exagerated Annexin response.  Does anyone out
>>there have any feedback on how trypsin affects Annexin staining?  Any
>>experience and or information about this will be greatly appreciated.
>>	Thanks in advance,
>>	Joan Kalnitsky
>>Joan Kalnitsky
>>Flow Cytometry Laboratory
>>VMRCVM
>>(540) 231-4115
>>FAX 540-231-7367
>>
>>	"It is better to serve than to receive"
>>	 B. Borg
>>
>>
>>	
>>
>>
>




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