[Cytometry] Mixing and matching brands

Juan Bolanos bolanosg.juan at gmail.com
Wed May 18 03:01:08 EDT 2011

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?


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
Kowloon Bay, Hong Kong

All that is gold does not glitter,**
Not all those who wander are lost;**
The old that is strong does not wither,**
Deep roots are not reached by the frost.*

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