Sarah
The proteinase K does a lot more than break the formalin linkages. To isolate
the RNA or DNA you have to break up the cell membranes, nuclear membranes and
all the other proteins in the cellular matrix to isolate the nucleic acids. I
don't think an antigen retrieval solution will do any
Hello
Just wanted to add one more thing - we actually use a dedicated pyrex dish
(maybe 6x10 inches) for our water bath for RNA sections. We use warm tap
water, but you can put it in the microwave for a short bit if it needs to be
warmer. You can spray the dish with RNAse away and wipe before
We have been using store bought gallons of distilled water in our water baths.
This water has been boiled so enzyme activity should be absent.
Helen
410.614.1660
http://tmalab.jhmi.edu/
http://prostatebiorepository.org/
Helen
-Original Message-
From:
We use a heat block during digestion. There is less chance of
contamination with a heat block than with a water bath.
... and no we don't use antigen retrieval solution for this! We use a
Qiagen kit too.
Mark
On Wed, Apr 3, 2013 at 9:11 AM, Helen Fedor hfe...@jhmi.edu wrote:
We have been
I attach my protocol for cell pellet
Preparation of formalin fixed Cell pellets.
1. Grow up approximately 5-1o millions cells.
2. Centrifuge the cells with the medium in conical tubes for 10 minutes with
speed 1000rpm.
3. Remove supernatant, wash once with 1X PBS at RT, centrifuge as before
Dear Colleagues
My current problem is: I should detect the myeloperoxidase (MPO) antigen and
its derivate nitrotyrosine and chlorotyrosine in the rabbit tissues (aorta) .
Most of the antibodies what I found and claimed to work on Rabbit are rabbit
polyclonal. Do you know antibodies for those
Histonetters,
Has anyone tried to process an intact mouse brain to paraffin?
Brett M. Connolly, Ph.D.
Principal Scientist, Imaging Dept.
Merck Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
brett_conno...@merck.com
T- 215-652-2501
F- 215-993-6803
Notice: This e-mail message, together
I've got a question regarding Ventana's Melanoma Cocktail. We've never done
double or triple staining at our institution so my knowledge of the CPT codes
for this is vague. If we were to use this product, which has HMB-45,
Tyrosinase, and Mart-1 in one dispenser, do I bill it as an 88342
You can only bill for 1
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
stephen.cla...@hcahealthcare.com
Sent: Wednesday, April 03, 2013 10:45 AM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet]
We have a vacancy for a Senior Histotechnian in our progressive department.
Experience in IHC a must. IF, ISH and /or EM desirable. Must be willing to
participate in and lead part of formal Continuing Education program for all
staff. Experience in QA development and Lean/Six Sigma projects a
Hello fellow histonetters ,
I need your expertise regarding the
treatment of nails. We currently immerse the nails specimen in a
solution of tween formalin up to 2 weeks . We then process and
section them. Is there a more rapid and efficient way to do it ???
Also,
we recently change
Use 10% aq. sol. of sodium or potassium hydroxide for 30 minutes on already
fixed toe (or finger) nails, wash thoroughly, and process as usual.
Decalcify BM with EDTA overnight
René J.
From: carmen loiselle carmen_loise...@hotmail.com
To: histonet@lists.utsouthwestern.edu
Hello All,
We section fixed frozen brain that we mount on the microtome using either 30%
sucrose or OCT and we keep the temperature down by surrounding the tissue with
crushed dry ice constantly (every 10 minutes).
One of the problems I have had, and also the person who trained me also had, is
Hello Maria,
Well, it's interesting that the Thermo HM 450 sliding microtome, looks very
much like the Microm HM450 sliding microtome
that I used (A LOT) at UCSF. You must be cutting (30um or 40um??)
free-floating sections.
Questions:
How thick (mm) are the frozen brain block?
AND, are you
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