[Histonet] re: iNOS
Kristopher Kalleberg
Kristopher.Kalleberg <@t> unilever.com
Tue Jul 26 13:29:43 CDT 2005
If I remember right Neomarkers/Lab Vision Corp. has iNOS antibodies. I used
them maybe 1 1/2 years ago but never got great results from the stain. I ran
the test numerous times and was unable to solve the problem. If you decide to
use Neomarkers and are able to achieve good results please send me your secret.
Good luck.
Kris Kalleberg
Histologist
Unilever R&D
40 Merritt Blvd.
Trumbull, CT 06611
203 381 5765
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Subject: Histonet Digest, Vol 20, Issue 35
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Today's Topics:
1. gluteraldehyde fixation with Re: [Histonet] Rat brains
(Gayle Callis)
2. IHC for RE:Staining for Helicobacter pylori (Gayle Callis)
3. Re: Staining for Helicobacter pylori (IamNinaOwen <@t> aol.com)
4. TMB staining for WGA-HRP (NIENHUIS,ROBERT )
5. historesin (Walters, Katherine S)
6. equivalent to Re: [Histonet] historesin (Gayle Callis)
7. Janella Seaton...please contact me (Morken, Tim - Labvision)
8. Background staining: mouse on mouse Broad spectrum kit
(Cathy Malcontenti-Wilson)
9. Re: historesin (Don.Birgerson <@t> leica-microsystems.com)
10. Re: CD 31 (Jackie M O'Connor)
11. North Carolina Histologist Position Available (MajorFocus <@t> aol.com)
12. Re: equivalent to Re: [Histonet] historesin (Pamela Marcum)
13. Looking 4 Antibodies (Luis Chiriboga)
14. DIF procedures (pathrm35 <@t> adelphia.net)
15. teaching heads for Nikon microscope (pathrm35 <@t> adelphia.net)
16. Integrase Interactor 1 (INI-1/hsnf5) antibody needed
(Histology SLU)
17. looking for iNOS antibody (Louro, Pedro)
18. Re: Integrase Interactor 1 (INI-1/hsnf5) antibody needed
(Dana Settembre)
19. IgG2a and IgG1 for guinea pig (Elizabeth Chlipala)
----------------------------------------------------------------------
Message: 1
Date: Mon, 25 Jul 2005 11:21:28 -0600
From: Gayle Callis <gcallis <@t> montana.edu>
Subject: gluteraldehyde fixation with Re: [Histonet] Rat brains
To: Laura Fidgen <lfidgen <@t> vt.edu>, Histonet <@t> lists.utsouthwestern.edu
Message-ID:
<6.0.0.22.1.20050725111447.01b7ada0 <@t> gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Question: is he planning to do EM on these tissues? If he had used
paraformaldehyde, your sectioning would be much easier - but if he is also
doing EM - you can try some things to counteract the dry/brittle brain.
In general, gluteraldehyde creates some friable, brittle tissues when
these are processed and embedded in paraffin. Check to see if you are
processing too long as long dehydrations, etc and heat from paraffin
contributes to the problem.
A good long soak in ice water or solid ice block with water on top after
trimming might help but be sure to not retrim the soak away, you have to
get the first sections off the knife. A warm water or room temperature
soak the going onto ice water for a time may help too, brain sometimes
needs a bit of help.
At 01:01 PM 7/23/2005, you wrote:
>Please help...
>A research project for our neuropathologist includes rat brains. These
>are extremely difficult to section as he profuses the whole rat with
>glutaraldehyde then takes brain samples. I have tried adding alcohol to
>my water bath but these sections still have many, many wrinkles. Can
>anyone offer suggestions? I would be so grateful for any and all advice.
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)
------------------------------
Message: 2
Date: Mon, 25 Jul 2005 11:25:52 -0600
From: Gayle Callis <gcallis <@t> montana.edu>
Subject: [Histonet] IHC for RE:Staining for Helicobacter pylori
To: Tom McNemar <TMcNemar <@t> lmhealth.org>,
Histonet <@t> lists.utsouthwestern.edu
Message-ID:
<6.0.0.22.1.20050725112230.01b7b5c8 <@t> gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Dear Tom,
If I had to do Helicobacter, I would vote for IHC too!!! Very specific for
identification of Helicobacter in human tissues, I only wished I had it for
murine animal model work but no antibody was available - boo hoo!!! There
is a nice method published in Histologic, Sakura Finetek website on doing
this IHC, and it was wonderful.
At 07:05 AM 7/25/2005, you wrote:
>We have been doing them by IHC for the last few years. Overkill, I know.
>The paths never like the Giemsa and we had so much trouble with the
>Warthin-Starry (consistency, repeats, etc) that I just switched to IHC.
>Works every time.
>Tom McNemar, HT(ASCP)
>Histology Co-ordinator
>Licking Memorial Hospital
>Newark, Ohio 43055
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)
------------------------------
Message: 3
Date: Mon, 25 Jul 2005 13:30:46 EDT
From: IamNinaOwen <@t> aol.com
Subject: [Histonet] Re: Staining for Helicobacter pylori
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <65.49f13cd4.30167bc6 <@t> aol.com>
Content-Type: text/plain; charset="US-ASCII"
Hi all
Thanks for all your views on this matter, keep the replies rolling in!
Nina Owen
------------------------------
Message: 4
Date: Mon, 25 Jul 2005 11:55:40 -0700
From: "NIENHUIS,ROBERT " <nienhuis <@t> ucla.edu>
Subject: [Histonet] TMB staining for WGA-HRP
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <1122317740.42e535ac936d3 <@t> mail.ucla.edu>
Content-Type: text/plain; charset=ISO-8859-1
We are about to cut a brain that was injected with
WGA-HRP for tract tracing. We haven't done this
for quite a while (decades?) and don't remember
if it is possible to delay processing after cutting.
If it is possible, how long can we delay, and what
solution should the cut tissue be stored in?
We plan to stain using the Olucha molybybdate
modification to the Mesulam TMB procedure.
Bob Nienhuis
UCLA / VA Medical Center
Los Angeles, CA
------------------------------
Message: 5
Date: Mon, 25 Jul 2005 15:07:50 -0500
From: "Walters, Katherine S" <katherine-walters <@t> uiowa.edu>
Subject: [Histonet] historesin
To: <histonet <@t> pathology.swmed.edu>
Message-ID:
<A4AA05CE92DACC43886D133DB94A926E0E995F4D <@t> medicine-exch1.medicine.uiowa.edu>
Content-Type: text/plain; charset="us-ascii"
I have a colleague looking to buy some historesin. Apparently it is
made by Leica (in Germany), and they will no longer ship it to the US.
Is there a company here that can sell it, or a similar product?
Thanks,
Kathy
------------------------------
Message: 6
Date: Mon, 25 Jul 2005 15:11:15 -0600
From: Gayle Callis <gcallis <@t> montana.edu>
Subject: equivalent to Re: [Histonet] historesin
To: "Walters, Katherine S" <katherine-walters <@t> uiowa.edu>,
Histonet <@t> lists.utsouthwestern.edu
Message-ID:
<6.0.0.22.1.20050725145504.01b275e0 <@t> gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
I don't recall plastic type of Historesin since it has been out of
circulation for a while in USA, but if this is GMA, buy Technovits 7100
If it is the methylmethacrylate, MMA then buy Technovits 9100.
Electron Microscopy Sciences sells both kits. These are excellent products.
At 02:07 PM 7/25/2005, you wrote:
>I have a colleague looking to buy some historesin. Apparently it is
>made by Leica (in Germany), and they will no longer ship it to the US.
>Is there a company here that can sell it, or a similar product?
>
>Thanks,
>Kathy
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)
------------------------------
Message: 7
Date: Mon, 25 Jul 2005 18:47:26 -0400
From: "Morken, Tim - Labvision" <tpmorken <@t> labvision.com>
Subject: [Histonet] Janella Seaton...please contact me
To: "'Histonet <@t> lists.utsouthwestern.edu'"
<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<0556BE8AC5551E4E8AF6BB9E42509BA20328CF55 <@t> usca0082k08.labvision.apogent.com>
Content-Type: text/plain
Janella, please contact me about the database you want.
From: Janella Seaton
----------------------------------------------------------------------------
----
Hello!
We want to implement a database for specimens we receive in our lab.
We would use this database starting with logging in specimens and ending
with archiving of specimens. It would be used to track when various steps
(gross photos, processing, staining, etc) have occurred and by whom.
We do want a system that is user friendly.
Any suggestions or advice?
Thanks for the help.....
j.s.
Tim Morken
------------------------------
Message: 8
Date: Tue, 26 Jul 2005 18:44:09 +1000
From: Cathy Malcontenti-Wilson <cmalc <@t> unimelb.edu.au>
Subject: [Histonet] Background staining: mouse on mouse Broad spectrum
kit
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<6.2.1.2.2.20050726182055.02ecbce8 <@t> mail.staff.unimelb.edu.au>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Dear All,
I really need some advice regarding an ongoing intense background problem.
We use FFP sections of mouse liver.
We are using an immuno kit called Zymed Histomouse Max Kit (It is a HRP-
DAB one).
Our primary ab is mouse anti human VEGF (1:1000).
The kit is called a BROAD SPECTRUM kit because it is meant to be able to be
used for mouse, rabbit, rat and guinea pig primaries.
The secondary antibody is not completely explained in the kit but I think
it is of goat host species.
Question: If it is broad spectrum does this mean that the secondary
antibody is a mixture of different antibodies?
The kit contains its own non specific block, with no details of what it
actually is.
Question: If the secondary ab is raised in goat, is this non specific block
normal goat serum? If not, what could it be?
We know that when I incubate the tissue with DAB only, we get endogenous
peroxidase staining which is pretty much abolished with H2O2 block.
We know that when we omit the primary antibody and keep all the other
layers, we get intense background staining of the sinusoidal (endothelial)
lining of practically all the sinusoids, and we get brown staining in
plasma cells, macrophages, connective tissue and other cells within vessels.
Question: Omission of non specific blocking reagent showed increased
background staining. Does this mean that the blocking agent is actually
blocking some of the background but not all? Why not?
We know that when we omit both the primary antibody and the non specific
block then we get background staining.
Question: What does this mean?
We know if we omit the secondary antibody only, we get no staining.
Question: What does this mean?
I would greatly appreciate any input into this problem which has been
plaguing us for a while now.
I hope someone can help, I cant beleive what a great wealth of knowledge
there is on this histonet list.
Cathy Malcontenti-Wilson
------------------------------
Message: 9
Date: Tue, 26 Jul 2005 07:32:06 -0500
From: Don.Birgerson <@t> leica-microsystems.com
Subject: Re: [Histonet] historesin
To: "Walters, Katherine S" <katherine-walters <@t> uiowa.edu>
Cc: histonet-bounces <@t> lists.utsouthwestern.edu,
histonet <@t> pathology.swmed.edu
Message-ID:
<OF3E1490E5.43943AC1-ON8625704A.0043C8DC-8625704A.0044E73B <@t> leica-microsystems.com>
Content-Type: text/plain; charset="US-ASCII"
Hi Kathy,
While we quit importing Historesin, there is product called
"Technovit " that is sold in the US by "Energy Beam Science" at
413-786-9322. If you have further questions, please call me.
Best regards,
Don
Don Birgerson
Leica Microsystems
Technical Assistance Center
Don.Birgerson <@t> Leica-Microsystems.Com
1-800-248-0123 ext 5918
"Walters, Katherine S" <katherine-walters <@t> uiowa.edu>
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
07/25/2005 03:07 PM
To
<histonet <@t> pathology.swmed.edu>
cc
Subject
[Histonet] historesin
I have a colleague looking to buy some historesin. Apparently it is
made by Leica (in Germany), and they will no longer ship it to the US.
Is there a company here that can sell it, or a similar product?
Thanks,
Kathy
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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Message: 10
Date: Tue, 26 Jul 2005 07:43:00 -0500
From: "Jackie M O'Connor" <Jackie.O'Connor <@t> abbott.com>
Subject: Re: [Histonet] CD 31
To: Angela McNabola <angela.mcnabola.b <@t> bayer.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<OF5BFE2A12.1853D9CE-ON8625704A.0045BBFE <@t> northamerica.intra.abbott.com>
Content-Type: text/plain; charset="us-ascii"
Santa Cruz gave me the same story - it's amazing that they depend on one
goat for a product line.
Jacqueline M. O'Connor HT(ASCP) QIHC
Assistant Scientist
Discovery Cancer R4N2 AP3
847-938-4919
Jackie.O'Connor <@t> abbott.com
Angela McNabola <angela.mcnabola.b <@t> bayer.com>
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
07/25/2005 05:40 AM
To: histonet <@t> lists.utsouthwestern.edu
cc:
Subject: Re: [Histonet] CD 31
Which antibody are you using? We have had great success with Santa Cruz
(goat
polyclonal) until I ordered a new lot this past month. When I called
them, they
indicated to me that they were having problems with their goat, literally.
I
guess it had stopped producing antibodies to CD31 that work in IHC
(interestingly enough, they still work in Westerns). They hope to have
the
problem resolved within the next month, with a new goat.
Angela McNabola, MS, HT(ASCP)SLS, QIHC
Bayer Healthcare
400 Morgan Lane
West Haven, CT 06516
"Kim Allred"
<kallred <@t> bellsouth.net> To:
<histonet <@t> lists.utsouthwestern.edu>
Sent by: cc:
histonet-bounces <@t> lists.utsouth Subject:
[Histonet] CD 31
western.edu
07/22/2005 03:15 PM
I am having a problem getting my CD31 to work. The protocol was for an
overnight incubation and pepsin pretreatment. I couldn't get it to work
this way so I altered it a bit by using trilogy for AR and a two hour
incubation for the primary ab. Does anyone have any ideas? This seemed
to
work well for a while but once again, I'm having problems with it. Any
input would be appreciated. Thanks in advance!
Kim Allred
Dermatopathology Associates
Jackson, MS
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------------------------------
Message: 11
Date: Tue, 26 Jul 2005 09:18:10 EDT
From: MajorFocus <@t> aol.com
Subject: [Histonet] North Carolina Histologist Position Available
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <b7.5c99a6d9.30179212 <@t> aol.com>
Content-Type: text/plain; charset="US-ASCII"
Frye Regional Medical Center in Hickory, North Carolina is currently
seeking a Histology Technician or Technologist for full time employment.
Qualified applicants must be certified by the ASCP or equivalent agency.
Please
apply on-line at _www.fryemedctr.com_ (http://www.fryemedctr.com/) or call
toll-free (866) 494-4747.
Frye Regional Medical Center
Human Resources
420 North Center Street
Hickory, NC 28601.
------------------------------
Message: 12
Date: Tue, 26 Jul 2005 09:25:08 -0400
From: Pamela Marcum <pmarcum <@t> vet.upenn.edu>
Subject: Re: equivalent to Re: [Histonet] historesin
To: Gayle Callis <gcallis <@t> montana.edu>, "Walters, Katherine S"
<katherine-walters <@t> uiowa.edu>, Histonet <@t> lists.utsouthwestern.edu
Message-ID: <6.1.1.1.2.20050726092213.019799b0 <@t> mail.vet.upenn.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Hi,
The Technovit 8100 is the same as JB-4 Plus which has a second polymer to
make the GMA remain clearer and somewhat less brittle. As Don was kind
enough to give you the number for EBS I shall not repeat and you can reach
Polysciences at 800-523-2575.
Pam Marcum
At 05:11 PM 7/25/2005, Gayle Callis wrote:
>I don't recall plastic type of Historesin since it has been out of
>circulation for a while in USA, but if this is GMA, buy Technovits 7100
>
>If it is the methylmethacrylate, MMA then buy Technovits 9100.
>
>Electron Microscopy Sciences sells both kits. These are excellent products.
>
>At 02:07 PM 7/25/2005, you wrote:
>
>
>>I have a colleague looking to buy some historesin. Apparently it is
>>made by Leica (in Germany), and they will no longer ship it to the US.
>>Is there a company here that can sell it, or a similar product?
>>
>>Thanks,
>>Kathy
>>
>>
>>_______________________________________________
>>Histonet mailing list
>>Histonet <@t> lists.utsouthwestern.edu
>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>Gayle Callis
>MT,HT,HTL(ASCP)
>Research Histopathology Supervisor
>Veterinary Molecular Biology
>Montana State University - Bozeman
>PO Box 173610
>Bozeman MT 59717-3610
>406 994-6367 (lab with voice mail)
>406 994-4303 (FAX)
>
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 13
Date: Tue, 26 Jul 2005 09:49:42 -0400
From: Luis Chiriboga <Luis.Chiriboga <@t> med.nyu.edu>
Subject: [Histonet] Looking 4 Antibodies
To: Histonet <Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <KFEIIJOCLBABEKFDAEHPAEFEFCAA.Luis.Chiriboga <@t> med.nyu.edu>
Content-Type: text/plain; charset="iso-8859-1"
HI everyone
I'm looking for antibodies against MLH-1, MSH-2, BRCA-1 and BRCA-2 for use
in human FF-PE tissue. recommendations as well as comments are welcome.
feel free to respond directly to my address.
Thanks in advance
Luis
------------------------------
Message: 14
Date: Tue, 26 Jul 2005 10:55:00 -0400
From: <pathrm35 <@t> adelphia.net>
Subject: [Histonet] DIF procedures
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<10787860.1122389700521.JavaMail.root <@t> web2.mail.adelphia.net>
Content-Type: text/plain; charset=utf-8
Fellow Techs,
Does anyone have Direct Immunofluorescence procedures they are willing to
share? We are interested in CD3, fibrinogen, IgA, IgM and IgG.
Thanks in advance.
Ron Martin, BS HT (ASCP) HTL
fax 561-721-1249
------------------------------
Message: 15
Date: Tue, 26 Jul 2005 11:01:06 -0400
From: <pathrm35 <@t> adelphia.net>
Subject: [Histonet] teaching heads for Nikon microscope
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<28528193.1122390066380.JavaMail.root <@t> web2.mail.adelphia.net>
Content-Type: text/plain; charset=utf-8
Fellow Techs,
Does anyone know of or where I can find binocular teaching heads for a (older
model) multiheaded Nikon Optiphot microscope? Any leads or info is greatly
appreciated.
Thanks in advance.
Ron Martin, BS HT (ASCP) HTL
fax 561-721-1249
------------------------------
Message: 16
Date: Tue, 26 Jul 2005 08:35:52 -0700 (PDT)
From: Histology SLU <sluhisto <@t> yahoo.com>
Subject: [Histonet] Integrase Interactor 1 (INI-1/hsnf5) antibody
needed
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <20050726153552.54597.qmail <@t> web51005.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Hello All:
I have a doc looking for the above antibody for use in FFPE tissue sections of
rhabdoid tumor in children. Any help would be greatly appreciated. Thanks, in
advance.
Susan
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------------------------------
Message: 17
Date: Tue, 26 Jul 2005 11:58:13 -0400
From: "Louro, Pedro" <pedro.louro <@t> spcorp.com>
Subject: [Histonet] looking for iNOS antibody
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<4508920F80C0D411B90200508BF9A9F4062B4A40 <@t> LAFMSG30.us.schp.com>
Content-Type: text/plain
Hi all,
I'm starting a new experiment with rat mesentery artery looking for
inducible nitric oxide synthase
(iNOS, eNOS, and nNOS).
What companies would carry this antibody that would work on FFPE rat tissue.
Also, what would be a good positive control?
Thanks in advance for your assistance
Pedro
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Message: 18
Date: Tue, 26 Jul 2005 12:13:31 -0400
From: Dana Settembre <settembr <@t> umdnj.edu>
Subject: Re: [Histonet] Integrase Interactor 1 (INI-1/hsnf5) antibody
needed
To: histonet <@t> lists.utsouthwestern.edu, sluhisto <@t> yahoo.com
Message-ID: <s2e62902.026 <@t> smtpnpc.umdnj.edu>
Content-Type: text/plain; charset=US-ASCII
I purchased it from BD Biosciences from San Diego, CA cat# 612110
I pretreat with DakoCytomation's Target Retrieval Solution. and use it
at 1:50 incubating for 15 minutes room temp.
Works nicely but there's not much in the vial and I am looking to Santa
Cruz, who sells 1ml of it.
Santa Cruz Cat. sc-13055
Good Luck.
Dana Settembre
University Hospital-UMDNJ
Newark, NJ
>>> Histology SLU <sluhisto <@t> yahoo.com> 7/26/2005 11:35:52 AM >>>
Hello All:
I have a doc looking for the above antibody for use in FFPE tissue
sections of rhabdoid tumor in children. Any help would be greatly
appreciated. Thanks, in advance.
Susan
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------------------------------
Message: 19
Date: Tue, 26 Jul 2005 10:48:10 -0600
From: "Elizabeth Chlipala" <liz <@t> premierlab.com>
Subject: [Histonet] IgG2a and IgG1 for guinea pig
To: "'Histonet'" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000101c59201$ce33a1c0$a7d48a80 <@t> AMY>
Content-Type: text/plain; charset="US-ASCII"
Hello everyone
I have been looking for some reagents for guinea pig for both ELISA and
IHC for IgG2a and IgG1. I did find a source for the IgG2a through
Nordic Immuno Reagents. Has anyone had any dealings with them? They
have several antibodies against guinea pig IgG2a all derived from
different species (sheep, goat and rabbit). I was guessing that the
rabbit antibody would be nice since I could use a polymer detection
system if necessary. Any suggestions of a different vendor or anything
else would be appreciated.
Thanks in advance.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz <@t> premierlab.com
www.premierlab.com
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309
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End of Histonet Digest, Vol 20, Issue 35
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