You can conjugate antibodies directly with different colors of Quantum dots using the conjugation kits from Quantum dots Co. The conjugation is not too difficult to follow the protocol that comes with the kits. Using biotin/streptavidin or secondary antibodies methods are often more difficult (if not possible) in multi-color staining than using the direct conjugation methods. Our experience of Qdots imaging indicated that higher concentration of Qdots-conjugated antibodies are often needed in intracellular staining than the use of the same antibody but conjugated with organic fluorochromes. Therefore, titration experiment with control samples are needed. Hope that helps. Jian ************************************* Jian Ling, Ph.D. Principal Engineer Bioengineering Section Southwest Research Institute 6220 Culebra Rd. San Antonio, TX 78238 Tel. (210) 522 - 3953 Fax. (210) 684 - 6147 Email. jling@swri.org Website: www.swri.org or www.bioengineering.swri.org ************************************* -----Original Message----- From: Alice L. Givan [mailto:Alice.L.Givan@Dartmouth.EDU] Sent: Friday, April 14, 2006 3:10 PM To: cyto-inbox Subject: Quantum Dots Hello, Can people tell me how you are staining cells with antibodies and the Quantum Dot fluorochromes? Are you conjugating the dyes to the antibodies yourselves (using reagents from where)? Or are you limiting yourselves to one Quantum Dot color with the biotin/streptavidin combination? Or are you using different animal-derived antibodies with animal-specific secondary antibodies (from where)? Or what am I missing? The goal is to use lots of QDot colors with just 407nm excitation -- but I don't know how close we are to being able to do this. Thanks. Alice Alice L. Givan Englert Cell Analysis Laboratory of the Norris Cotton Cancer Center Dartmouth Medical School Lebanon, NH 03756 USA tel 603-650-7661 fax 603-650-6130 givan@dartmouth.edu www.dartmouth.edu/~celllabReceived on Tue Apr 18 13:38:00 2006
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