detection of cytoplasmic IgM

mscolnik@interlink.com.ar mscolnik at interlink.com.ar
Tue Dec 8 14:32:19 EST 1998



----------
>From: Tsurusawa Masahito <tsuru at aichi-med-u.ac.jp>
>To: Cytometry Mailing List <cytometry at flowcyt.cyto.purdue.edu>
>Subject: detection of cytoplasmic IgM
>Date: mart., 8 dici 1998 11:18
>

>
>Hallo Flowers,
>
>How do I detect cytoplasmic IgM(m chain) in pediatric preB -ALL cells
>together with other cytoplasmic
>
>antigens?    Until now I have used saponin solution to detect
>cytoplasmic m chain in childhood ALL.
>
>This method was excellent although the cell aggultination sometimes
>occured for poor marrow samples.
>
>Now I am begining to detect  other cytoplasmic antigens of CD79a, CD3
>and MPO
>
>by using the commercial permeabilization kits ( FACS permeabilizing
>solution, Coulter Intraprep,
>
>DAKO intrastain, Fix & Perm), but these methods did not refer to the
>detection of cytoplasmic m chain
>
>in the commercial brochure (or even in the cyometry journals).   Since
>the pediatric marrow sample was
>
>so small, I want to measure the above four intracellular antigens by a
>single permeabilizing method.
>
>Thanks
>
>
>Masahito Tsurusawa MD
>
>associate professor of pediatrics,
>
>Aichi Medical University, Aichi, 480-1195, Japan
>
>

Dear Masahito:
In our lab, usually use the Permeafix (Ortho) permeabililization solution
for all cytoplasmic antigens for diagnosis of oncohaematological diseases
without problems. The differences between the saponins and some commrcial
kits is the alteration of SSC characteristics of the blast cells.
Is very important (in our experience) for  the determination of cytoplasmic
IgM the control negative.
Usually we use a tube with IgA or IgG for determinate the control negative.

Best wishes

Mariano Scolnik MD
IO/IIHEMA
Academia Nacional de Medicina de Buenos Aires
mscolnik at interlink.com




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