[Histonet] Re: CY2/CY3 vs Alexa488/546-555
To get around the problem of Alexa conjugated secondaries you don't like
from Molecular Probes, buy your favorite secondary from Jackson, and
conjugate it yourself with an Alexa dye conjugation kit from Molecular
Probes. You will get the best of both (of your favorite) worlds. The kit
conjugates three antibodies, and is a very simple to use. This may be the
only way to guarantee the quality you desire and still have your favorite
We have experienced similar problems as you with MP Alexa conjugated
secondaries. However, last week's IFA using a highly adsorbed goat
antiRabbit-Alexa 488 at 0.5ug/ml with a rabbit polyclonal for a bacteria in
frozen mouse lung sections gave spectacular results - a pleasant surprise
for me and one PI.
At 05:40 PM 12/21/2005, you wrote:
>Has anyone done a SIDE-BY-SIDE comparison of Cy2 or Cy3 as compared with
>Alexa 488 or Alexa 546/555? I am ideally looking for experimental results
>not anecdotal information if possible .... I know we have all heard that
>Alexas are the best and this is certainly true as compared to FITC. But
>are they better compared to CY2 and CY3 from Jackson Labs?
>I ask b/c I love Jackson Labs secondaries and they don't sell Alexa
>conjugates :( I use Alexa Streptavidin from Molecular Probes but am not
>such a fan of their secondaries for TISSUE staining (they give me some
>strange artifacts sometimes although they work great for cell culture
>work). I want to keep ordering Jackson ImmunoResearch quality
>secondaries - they are just SO SPECIFIC for mouse work but they only sell
>CY2 and CY3 (which have worked for me thus far).
>Decisions, decisions .... Comments, opinions, suggestions?
Gayle Callis HTL, HT, MT(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717
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