Multiplex Bead Assays

Moss, Delynn M. (CDC/CCID/NCZVED) dmm3 at CDC.GOV
Tue Jul 17 06:38:29 EDT 2007


Andrew,
 
The protocol for antibody coupling to COOH beads at the Luminex website
should work.  If you can not get in, give them a call.	If that does not
work, I'll send you a protocol.
 
A 635 nm laser will excite the two dyes in the Luminex beads, but I do
not believe you will be able to distinguish very many bead
classifications.  The Luminex instrument can distinguish up to 100
different Luminex bead classifications, and if you use these beads, I
suggest you use a Luminex instrument.
 
Another option is Bangs Laboratories.  They have COOH beads that are
excited by 488 nm laser with emission at about 675 nm, but there are
only five different bead classifications, resulting in only five-analyte
detection.  I have used these beads, and there is no problem using PE or
FITC as the reporter fluorochrome.  I believe BD has an array of eight
beads that work in the same fashion as the Bangs beads, but I have no
experience with the BD kit.
 
Hope this helps.
 
Delynn M. Moss 
Centers for Disease Control 
CCID/NCZVED/DPD/PDB 
4770 Buford Hwy, NE 
Atlanta, GA 30341 
ph 770-488-4041 
fax 770-4884108 
e-mail dmm3 at cdc.gov 

   

________________________________

From: Andrew Beernink [mailto:Andrew.Beernink at dako.com] 
Sent: Wednesday, July 11, 2007 12:09 PM
To: cyto-inbox
Subject: Multiplex Bead Assays


Hello-
 
I have a client who wants to set up multiplex bead assays for soluble
analytes.
 
Some of the commercial kits will prove useful, but said client would
also like the ability to use his own antibodies.
 
1.  Can anybody provide a protocol for conjugating Ab to beads?
Spherotech was kind enough to provide some bead samples free of charge
(with and without a carboxyl moiety).  Luminex has a protocol on their
website, but the link is faulty (as are a LOT of their links).
 
2.  Will Luminex bead sets work on current 405/488/635 cytometers?
Alas, links to their instrument specs are also a bit whacked.
 
Thanks for any feedback.
 
Andrew
 
Andrew Beernink
Senior Applications Specialist
Dako Colorado
(858) 353-7007
(858) 435-1137 efax
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