I'm looking to contact Rebecca (Becky) Orr. Does anyone know her current email
address? The one I have bounced back.
Thanks, Andrea
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Hi All, Do you have a protocol for formalin fixation (and processing to FFPE)
of a previously snap frozen piece of tissue? Or advice as to the best method of
snap freezing for such a process (assume isopentane slurry)? We have done this
for several tissues with our standard fixation and processi
Just curious as to at what shelf-life people consider their blocks too "old" to
rely on for IHC data? Along those lines I am wondering about labile antigens
... does anyone know how fast epitopes may "disappear" from blocks and what are
some good examples of labile epitopes/antigens? As usual an
Congratulations Dr. Adam on your recent defense (yay). Thanks for the kind
words, I have enjoyed emailing with you and discussing bone IHC. We will miss
you on Histonet and I certainly hope you return to research once you have
completed your clinical years (if not sooner).
Best, Andrea
_
Hi All,
I am wondering if anyone knows of any particular publications or studies that
have examined manners in which one can predict whether an antibody in a large
panel may work in IHC. Let's say you are faced with 50-100 Abs and you need to
determine which works in IHC, but all you know is bi
Is anyone currently staining for RFP in FFPE or frozen sections using a
detecting antibody? I am curious as to what anti-RFP Ab would be recommended.
There is one I plan to try from Invitrogen (rabbit anti-RFP) but of course
would love feedback if people have used one in particular.
Thanks, An
Agree, switching to Leica with disposable blade holder changed my life. Never
looked back. Love the CM3050S.
--- On Sun, 7/10/11, Rene J Buesa wrote:
From: Rene J Buesa
Subject: Re: [Histonet] In search of a cryostat...
To: "histonet@lists.utsouthwestern.edu" ,
"Tom McNemar"
Date: Sunday,
It will depend on how your scope is configured. On our system, when using the
three you have outlined I routinely use CY5 or Alexa633 (or equivalent in far
red category) on our scope. It's actually my personal favorite. If you have any
doubts check out the spectral viewer on Invitrogen and also
Hi All,
I am doing a survey and will be happy to compile results and share if folks
will respond! What is your favorite antigen retrieval method and/or panel?
Buffer
Source/composition
Temperature
Device
Thanks, Andrea
___
Histonet mailing li
Use the TUNEL assay, several very easy to use kits available. Roche's Cell
Death Detection kit with FITC might be the easiest.
--- On Thu, 4/1/10, Alexandra Meinl wrote:
From: Alexandra Meinl
Subject: [Histonet] Assay for cell death needed
To: "Histonet"
Date: Thursday, April 1, 2010, 9:00
Very interesting! Coming out the nose is definitely bad for any work I have
done in the past - lungs get blown out, liver doesn't perfuse well and bone
marrow looks horrific. However, if you are working with PFA and doing a
post-fix anyway, you will probably be fine. If you are using GA and coun
We used to do overnight in our DAKO Autostainer many years ago, and we kept
many containers of dH20 in the incubator to make a humid chamber. Seemed to
work well - although we realized overnight was overkill and 1-2 h was
sufficient. In fact, given this we started to always put humid chambers in
aries in 0.2X concentration of your block.
If you want to share your protocol, maybe I can give you some further tips?
Let me know if I can help further,
Andrea
--- On Wed, 3/3/10, Mauricio Avigdor wrote:
From: Mauricio Avigdor
Subject: Re: [Histonet] IF staining on peritoneal macrophages
overkill.
Let me know if you need anything else,
Andrea
--- On Wed, 3/3/10, Mauricio Avigdor wrote:
From: Mauricio Avigdor
Subject: Re: [Histonet] IF staining on peritoneal macrophages
To: "Andrea T. Hooper" ,
histonet@lists.utsouthwestern.edu
Date: Wednesday, March 3, 2010,
. I
would give you the catalog number except I am not at my computer.
Andrea T. Hooper
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
. I
would give you the catalog number except I am not at my computer.
Andrea T. Hooper
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
t: Re: [Histonet] Snap Freezing Tissue
To: "Andrea T. Hooper"
Cc: histonet@lists.utsouthwestern.edu, "Laurie Colbert"
, "Steve Pike"
Date: Wednesday, February 24, 2010, 10:31 PM
I learned to freeze tissue by taking a beaker and filling it with
2-methylbutane and t
rm storage.
If snap freezing tissue for RNA/protein extraction I use the classic isopentane
cooling method.
Andrea T. Hooper
--- On Wed, 2/24/10, Laurie Colbert
wrote:
From: Laurie Colbert
Subject: [Histonet] Snap Freezing Tissue
To: histonet@lists.utsouthwestern.edu
Cc: "St
Not sure if you were asking me or not and it's a broad answer but VE-cadherin
is a good one - the best overall marker of endothelium. vWF is another one that
works well for most tumors. If staining mouse organs, if you let me know what
tissue you are staining I can recommend the best specific ma
What types of mouse tissue are you working with? Are they fresh frozen with
post-fixation or fixed frozen? Do you block biotin? Do you have an isotype
control and a no primary control? I am thinking biotin's the likely culprit as
the kit doesn't come with the very necessary biotin blocking reag
to mention the autofluorescence from
RBCs. I am afraid of false positives.
--- On Tue, 2/23/10, Adam . wrote:
From: Adam .
Subject: Re: [Histonet] Double labeling with antibodies that need different
fixatives
To: "Andrea T. Hooper"
Cc: "Phebe Verbrugghe" ,
histonet@lists.utsouthwes
Hi Phebe,
In my experience that CD31 clone you refer to doesn't work well mainly b/c of
paraffin embedding - in combination with PFA/formalin fixation. On
PFA/formalin fixed frozen samples it works just fine with a trypsin antigen
retrieval step. I find it's the paraffin that is the real kill
Although what you say is true, it is worth noting that there are some
antigens/antibodies more amenable to acid decal. I have experienced this myself
several times recently and been burned by thinking EDTA will always give better
IHC results. At least for murine bone marrow vasculature antigens,
Hi Julia,
I am sorry it's taken me months to reply to this email and hopefully it's not
too late. I am only catching up on Histonet messages recently.
I stain for VE-cadherin, CD144 in murine bone routinely. R&D Systems goat
anti-mouse VE-cadherin works very well in paraffin and frozen murine
After cytospinning I always dry my slides thoroughly before moving on to a IHC
or IF step. I also use Superfrost Plus slides or silane coated slides to
increase adherance. I would do that then try various fixatives.
You can make a cell pellet as you suggest. Decide ahead of time if you want to
For many years doing IHC on mouse bones, I have used 10% EDTA, just pHed up to
where it goes into solution, with great results for frozen section
immunofluorescence. It takes 5 days. For FFPE, we use Richard Allan Decal
solution which is HCl and EDTA. Also works great and only takes 1 hour for
Couldn't agree with Gayle more!
Glucose oxidase is the best protocol for blocking endogenous peroxidase but
isn't entirely necessary unless you are working with hematopoietic tissue or
tissue that has a large leukocyte infiltrate. For muscle standard 0.3%
H202 should be fine. Of course if you
Your protocol is fine. It's precisely my protocol.
What do you get when you do the no primary control? Although this control isn't
sufficient for publication it is necessary for troubleshooting. In fact
performing the following controls together can tell you exactly where the
problem lies:
Hi everyone! It's good to be back ... I have resubscribed under a different
email as I switched place of employment.
I am just running a poll as to what your favorite anti-VEGFR2 antibody is for
immunostaining human tissue or cells. Frozen or paraffin, cells or tissues
... it's all good info
Whether or not an antibody can be used for western vs IHC depends on
many factors.
Some antibodies can be used for both, some can be only used for one
vs another, and some cannot be used for either.
There are quite a few factors to consider and here are a few from the
top of my head (and I a
30 matches
Mail list logo
