[Histonet] Procedure for manual C4D immunofluorescence staining

[James L Burchette]

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

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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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