Just to add to Linda's note on how we do our C4d's, our control slides are cut in advance, fixed for 10" in acetone and stored in the freezer.  We currently use previously tested positive patients.

Sally Ann Drew-MT(ASCP)
Univ. of Wisconsin Hospital & Clinics
IHC/ISH Laboratory 
600 Highland Ave. VAH-DM223
Madison, WI 53792-2472
608-265-6596
sa.drew@hosp.wisc.edu


-----Original Message-----
From: Sebree Linda A. 
Sent: Thursday, January 30, 2003 11:14 AM
To: Rita Humphrey
Cc: Histonet
Subject: RE: C4d staining for renal biopsies


Hi Rita and histonetters,

OK, here's the scoop on our C4d staining of renal biopsies:

Biogeneis C4d catalog #2222-8004
10" R.T. acetone fixation of 4um thick frozen renal biopsies, air dry.
Stain on the Ventana ES for 16" or the Ventana NexES for 32" at 1:600.
Use Ventana iview DAB detection, Ventana AB block (biotin block) and hematoxylin counterstain.

We are currently investigating the use of Ventana's Morphosave as a 2" pretreatment in order to clean up the stain a bit.  It helps on the unusually dirty tissues but does compromise the morphology a bit.  The jury is still out on this.

Linda A. Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-2472
600 Highland Ave.
Madison, WI 53792
(608)265-6596
FAX: (608)262-7174


-----Original Message-----
From: Rita Humphrey [mailto:Rita.Humphrey@chsys.org]
Sent: Thursday, January 30, 2003 9:48 AM
To: Sebree Linda A.
Subject: RE: C4d staining for renal biopsies


My pathologist just ask me to order this antibody, would you please fax me
you procedure? Fax 205-939-6869 Thank you, Rita

-----Original Message-----
From: Sebree Linda A. [mailto:la.sebree@hosp.wisc.edu] 
Sent: Friday, September 27, 2002 12:02 PM
To: Martha Ward
Cc: Histonet
Subject: RE: C4d staining for renal biopsies


Martha,

We only do C4d on frozen renal biopsies  mainly because the clinicians want
their answer sooner rather than later.

Linda A. Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-2472
600 Highland Ave.
Madison, WI 53792
(608)265-6596
FAX: (608)262-7174


-----Original Message-----
From: Martha Ward [mailto:mward@wfubmc.edu]
Sent: Friday, September 27, 2002 10:22 AM
To: HISTONET@pathology.swmed.edu
Subject: C4d staining for renal biopsies


Our lab has had inquiries about adding this antibody to our protocol for
transplant renal biopsies and I wanted to hear from labs that are currently
performing it.  All the protocols I have seen reference frozen tissue but we
were wondering if there is a way to do it on ffpe tissues.  Any and all help
will be greatly appreciated!
 
Martha Ward, MT (ASCP) QIHC
Wake Forest University Baptist Medical Center
Winston-Salem, NC
1-336-716-2756