APC-Cy7 conjugated Streptavidin

Samantha Bailey sam-bailey at northwestern.edu
Tue Nov 23 17:23:40 EST 2004


Pamela,   does she see the same thing with other conjugated 
streptavidins?	The RAW cells will be extra sticky (macs), will 
non-specifically bind products.  try blocking with 20% autologous serum 
along with the usual FcgR antibody.

Are you also staining with APC?  the background may be bleed over from 
the APC channel.  Also, activated macs will flouresce in the APC 
channel, so you may be picking this up too.  PE-Cy7 has less bleed over 
into APC, & may work better for you.

Best of luck,

Sam

On Nov 22, 2004, at 10:56 AM, Pamela Shaw wrote:

> Hi all,
>
>  
>
> I have an investigator who is having problems with non-specific 
> binding when using eBioscience APC-Cy7 conjugated Streptavidin.  
> (Catalog # 10-4317)  She is staining RAW 264.7 cells.  She gets 
> non-specific binding with or without the addition of CD40 biotin.  She 
> has tried several concentrations, and all seem to have this 
> non-specific binding.  Has any one else had problems with this 
> particular product, or have any suggestions for us?
>
>  
>
> Pam Shaw
>
> Fluorescence Cytometry Core Facilitator
>
> Center for Environmental Health Sciences
>
> The University of Montana
>
> 32 Campus Dr, Skaggs Building 052
>
> Missoula, MT 59812
>
>  
>
> Ph (406) 243-4545
>
> email:  pamela.shaw at umontana.edu
>
> www.umt.edu/cehs
>
>  
>
Samantha L. Bailey, PhD
Northwestern University
Feinberg School of Medicine
Department of Microbiology / Immunology
Chicago, IL 60611

sam-bailey at northwestern.edu
Phone: 312 503 1449
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