[Histonet] Procedure for manual C4D immunofluorescence staining

[Anne van Binsbergen]

thanks for this method
we have been battling to get C4d FITC to work
hopefully i can get a vendor here in the desert to source the reagents (that
is a huge problem here)
AbuDhabiAnnie

2009/10/16 James L Burchette <burch007 <@t> mc.duke.edu>

> Frozen sections cut at 3-4 microns. Air dry 30 minutes.
> Acetone fix for 5 minutes (room temp acetone), air dry slides
> PBS wash x2
> C4d (diluted 1:500 in 1% BSA/PBS) incubated 1 hour. Mouse monoclonal C4d
> is from AbDirect/Serotec
> PBS rinse, wash x2
> Cy2 labeled goat anti-mouse IgG (diluted 1:100 in 1% BSA/PBS), incubated
> 30 minutes (Cy2 GAR from Jackson ImmunoResearch Labs)
> PBS rinse, wash x2
> Coverslip with fluorescence mounting media
> Happy viewing
>
> I run this protocol everyday!
>
> Jim Burchette, HT(ASCP) QIHC
> "A simple histotech from a little country hospital in North Carolina"
>
>
>
> "Foshey, Annette" <AFoshey <@t> chw.org>
> Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
> 10/15/2009 04:56 PM
>
> To
> " (histonet <@t> lists.utsouthwestern.edu)" <histonet <@t> lists.utsouthwestern.edu>
> cc
>
> Subject
> [Histonet] Procedure for manual C4D immunofluorescence staining
>
>
>
>
>
>
> Histonetters,
> Does anyone have a procedure they could share for manual C4D
> immunofluorescence staining?
>
> Thanks in advance
>
> Annette Foshey, HT (ASCP)
> Charge Tech in Histology
> Children's Hospital of Wisconsin
> 414-266-6580        Fax 414-266-2779
> afoshey <@t> chw.org
>
>
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-- 
Anne van Binsbergen (Hope)
Abu Dhabi
UAE