[Cytometry] Mixing and matching brands

Mikhail Roshal mir9079 at med.cornell.edu
Wed May 18 09:39:20 EDT 2011


H Juan,
You can use Abs from different manufacturers as long as the Ab itself  
performs well.   It is done routinely.   That being said conjugation  
efficiency and the stability of tandems may vary from one manufacturer  
to another and for that matter from one lot of the same manufacturer  
to another.
Like for all tandems you will need to do your compensations on the  
specific antibody you are using for the specific lot number you are  
using and taking care of the Ab stability. I have no experience with  
the specific antibody in question.
Misha Roshal
Assistant Professor of Pathology and Laboratory Medicine
Weill-Cornell Medical College.
On May 18, 2011, at 3:01 AM, Juan Bolanos wrote:

> Hello Everybody!
> I am well on my way to my 4th month of self teaching FLow Cytometry  
> and this
> forum, as well as Mario Roeder's webpage (and many other sources)  
> have been
> wonderful in this process. I have currently completed my 4 color  
> panel for
> Amnion Stem Cells and all is good, however i am now running into a  
> bit of
> a conundrum. I am in need of adding a 5th color as I am attempting to
> measure Ep-CAM as well as my other  4 dyes. I am using AF488, PE, PI,
> PerCP/Cy5.5 and want to incorporate PE/Cy7 for EpCAM, however the  
> company
> that supplies my antibodies (Biolegend) does not have Ep-CAM-PE/Cy7.  
> I have
> attended some flow seminars and I noted at one Beckman Coulter  
> conference
> that they did not recommend the use of different brands of  
> antibodies within
> the same panel, as they claim the different manufacturing processes  
> may
> affect the fluorescence and binding properties of the AB's. To a  
> certain
> extent this makes sense, however it sounds like a ploy for you to  
> get stuck
> with a single brand of manufacturer, as I think that proper  
> compensation
> will make up for any discrepancies amongst the different brands.
> My question would be: Has anyone had any adverse effects when mixing  
> and
> matching conjugated antibody brands?
>
> Also, my other alternative would be to purchase Qdots (Qdot 800) from
> BioLegend, however they seem to have just begun to enter the Qdot  
> market and
> i do not know their performance (as well as the fact that i have never
> previously used a Qdot). Has any one used Biolegend Qdots before?
> Thanks!
>
>
> -- 
>
> Thank you for your time and patience and happy year of the rabbit!
>
> Best Regards
> BSc in Biotechnology
> Juan Bolanos
> ProStemCell Technologies
> #1 Trademart Drive, Hong Kong Kowloon Bay International Trade and  
> Exhibition
> Center
> Kowloon Bay, Hong Kong
>
> ____*
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> Not all those who wander are lost;**
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> Deep roots are not reached by the frost.*
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