Re: MIB1
<< Previous Message | Next Message >>
From: | Mary Vaughan <mvaughan@sc3102.med.buffalo.edu> (by way of histonet) |
To: | histonet <histonet@magicnet.net> |
Reply-To: | |
Content-Type: | text/plain; charset="iso-8859-1" |
Dear Heike,
MIB-1 [Ki-67] took a little extra time.... but I managed to get the
assay working. I've not done bone marrow, but even "regular" soft tissues
[form fix - paraffin] required an extra effort. The literature pointed in
this direction also, although I don't have the references at my fingertips.
Temperature and duration are two very important factors here. This
is the only Ab [in my lab] that requires double time in the microwave
{citrate pH 6.0}. My usual pre treatment time is 10 min and with this
Antibody, 2 X 10 min worked nicely. Also, I used an overnight incubation
with the primary @ 4 µg/ml. I tried 6 different pre treatments, and nothing
else worked. {Don't forget your negative control}. Your secondary system
sounds OK. Let me know if you have any specific questions.
At 10:14 PM 3/4/99 -0800, you wrote:
>Histonetters!
>anybody out there doing immunohistochemistry on bone marrow sections with
>MIB 1? We get no staining so far. However, I have to admit that in
>Germany the routine fixation for bone marrow biopsies contains a
>combination of glutaraldehyde, formaldehyde and other ingredients. So, I
>currently assume it is the glutaraldehyde which destroys the MIB1
>epitope.
>But may be, there are some suggestions aout there? some special
>pre-treatment? We use a pressure cooker (5 min) in citrate buffer at the
>moment and a straeptavidin-biotin-hrp- detection system.
>
>thanks for your help
>
>Heike Grabsch, Germany
>
>
Best Regards,
Mary Vaughan HT(ASCP)
Roswell Park Cancer Institute
Pharmacology + Therapeutics
Elm + Carlton Sts. CDC-121
Buffalo, NY 14263
<< Previous Message | Next Message >>