[Histonet] RE: Endogenous Tau (normal Tau)

2014-11-14 Thread Hoekert, W.E.J.
Hi Claudia, We use Dako anti-Human Tau (rabbit polyclonal) at a concentration of 1:8000. We do not use a pretreatment (HIER or protease). Staining is done on a Ventana stainer. Good luck, Willem Van: histonet-boun...@lists.utsouthwestern.edu [histonet

RE: [Histonet] Corpora amylacea are destroyed after antigen retrieval?

2014-02-20 Thread Hoekert, W.E.J.
I am not sure if you are talking about the Beta Amyloid antibody, but if you are: the best way to do the antigen retrieval is 3 minutes in formic acid. Willem Van: histonet-boun...@lists.utsouthwestern.edu namens Pirici Daniel Verzonden: di 18-2-2014 23:59 Aan

RE: [Histonet] M2D2 and SMAD

2013-12-05 Thread Hoekert, W.E.J.
Hi, SMAD4 is very difficult on the Ventana platform. The only clone that would work on the Ventana to my knowledge(clone EP618Y, immunologic) has recently 'collapsed' (i hope that's the right word?) It means that it is not available anymore. Maybe there is some vendor that has a few vials left

RE: [Histonet] Her2 Dual ISH and breast processing

2012-12-14 Thread Hoekert, W.E.J.
is: http://www.histosearch.com/rene.html >From: Rene J Buesa >To: "Hoekert, W.E.J." ; "vtol...@cox.net" >; >"histonet@lists.utsouthwestern.edu" > [] >Willem, yes, there are. Look through the attached references which include >some fo

RE: [Histonet] Her2 Dual ISH and breast processing

2012-12-13 Thread Hoekert, W.E.J.
I was wondering,is there any literature on this subject? i.e. the minimal required fixation time of breast tissue in order to get reliable immuno staining (Oestro, Prog and Her2neu and ISH). We immunologists are alway trying to convince our pathologists about the importance of good fixation, but

RE: [Histonet] MSI antibodies

2012-11-30 Thread Hoekert, W.E.J.
Hi Kenia I guess you will need at least two blocks because they are never 'positive' all four of them in the same tumour. You might search your archive for young people with colon cancer. There is a good chance that you will find positive material. Willem Hoekert OLVG The Netherlands ___

RE: [Histonet] Pronase antigen retrieval

2012-09-20 Thread Hoekert, W.E.J.
Hello Eva, We are not using pronase anymore, but when we did, we were doing it at room temperature. (15 mg in 250 ml PBS, sigma P-5147). It worked fine. Our Trypsin antigen retrieval, we were doing it at 37C as follows: the evening before the retrieval, we put some PBS in the 37C stove, the

[Histonet] How to make HP control tissue?

2012-07-04 Thread Hoekert, W.E.J.
Hi Histonetters, Does anybody has experience in making your own HP control tissue? I am tired of using positive biopsies of patients since they are always almost finished. I have heard of a procedure on making HP controls but I am not exactly sure how it is done. I have tried the following:

[Histonet] MDM-2 on Ventana, which clone?

2012-06-01 Thread Hoekert, W.E.J.
Hi Histonetters, I am having a very hard time in setting up the MDM-2 antibody on our Ventana Benchmark XT's. I am using clone SMP14 from Becton Dickinson, which was working fine on the Labvision. I have tried everything (even incubating overnight at 4ºC) and I am out of idea's. Maybe I should

RE: [Histonet] SMAD4

2012-04-25 Thread Hoekert, W.E.J.
Which stainer are you using? Because some clones will not work on a Venatana stainer. Willem Van: histonet-boun...@lists.utsouthwestern.edu namens Chakib Boussahmain Verzonden: ma 23-4-2012 4:20 Aan: histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] SMAD4

RE: [Histonet] IHC and negative controls?

2012-02-17 Thread Hoekert, W.E.J.
We are not using negative controls. We only add positive controls at the bottom of our patient slides. Sometimes there are 3 different types of control tissue (a TMA), so that you can use the same control block for several antibodies. You could add a negative control in the same block as the pos

RE: [Histonet] Pms 2 antibody trouble

2012-01-13 Thread Hoekert, W.E.J.
This is what we are doing on our Benchmark XT's: MLH1: cc1 30', 32' incubation time (Ventana Cell Marque) MSH2: cc1 60', 60' incubation time (Ventana Cell Marque) MSH6: cc2 60', 60' incubation time (1:600, Becton Dickinson, clone 44) PMS2: cc1 30', 32' incubation time + amplification (1:50, Becto

RE: [Histonet] HSV1 and HSV2

2011-12-27 Thread Hoekert, W.E.J.
For the immunologic pAB cocktail we are using Protease 1 (4 min) and ab incubation of 32 min. The only problem is that mast cells are also staining, it is a side effect of the protease 1. (AB concentration 1:400) Willem Hoekert OLVG Amsterdam The Netherlands __

RE: [Histonet] GLUT-1 antibody

2011-11-29 Thread Hoekert, W.E.J.
Hi Sandra, I am using Neomarkers (Glut 1 Ab-1, polyclonal) on the Benchmark XT. 30' cc1 32' ab incubation 1:50 It works fine. Willem Hoekert OLVG Amsterdam The Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens Diaz, Sandra Verzonden

RE: [Histonet] Fatty mamma tissue keeps on washing off

2011-11-17 Thread Hoekert, W.E.J.
slides. So from now on, if we have a case that washes off, we will be using the Dako slides. Thanks for your help, Willem Hoekert OLVG AMsterdam The Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens Hoekert, W.E.J. Verzonden: do 20-10-2011

[Histonet] Fatty mamma tissue keeps on washing off

2011-10-20 Thread Hoekert, W.E.J.
Hi Histonetters, Every once in a while, we have mamma tissue that washes off the slides completely. The reason for this is that the tissue is still too fat and/or not sufficiently fixed. There is no problem with the HE stain, but the immuno will wash off. What do you do in such cases? I have t

RE: [Histonet] P 16

2011-09-30 Thread Hoekert, W.E.J.
Hi Phyllis, We are doing a double stain (Mib-1 - P16) on cervix biopsies in which the P16 antibody is coloured with AP. It works fine, our pathologists are quite fond of it. Almost every day we have a few slides. Willem Hoekert OLVG, Amsterdam The Netherlands __

RE: [Histonet] Microsatellite Instability Antibodies

2011-03-24 Thread Hoekert, W.E.J.
>We are currently running the Ventana IHC stainers with the UltraView Detection. Just like us! MLH-1: Cell Marque (ref 760-4264). 30' cc1, 32' ab incubation. MSH-2: Cell Marque (ref 760-4265). 60' cc1, 60' ab inc. PMS2: Becton Dickinson, clone A16-4, cat nr 556415. 30' cc1, 32' ab inc. +amp

RE: [Histonet] RE: Procedure for making Gram Control

2011-03-18 Thread Hoekert, W.E.J.
Thomas Jasper Verzonden: do 17-3-2011 18:32 Aan: Hoekert, W.E.J. CC: histonet@lists.utsouthwestern.edu Onderwerp: RE: [Histonet] RE: Procedure for making Gram Control Some sort of small, snack sausage of questionable quality. I'm not familiar with anything like that in Europe, but maybe you

RE: [Histonet] RE: Procedure for making Gram Control

2011-03-17 Thread Hoekert, W.E.J.
What would be the equivalent of a Slim Jim in Europe? The Netherlands to be more precise? Van: histonet-boun...@lists.utsouthwestern.edu namens Walter Benton Verzonden: wo 16-3-2011 15:07 Aan: Hayes, Randi (HorizonNB); histonet@lists.utsouthwestern.edu Onderw

RE: [Histonet] ER staining in Tonsil.

2010-12-10 Thread Hoekert, W.E.J.
Yes, we see the same. We also see staining in follicles in appendix and colon. Van: histonet-boun...@lists.utsouthwestern.edu namens Andrea Verzonden: do 9-12-2010 15:05 Aan: Histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] ER staining in Tonsil. Good

RE: [Histonet] Formalin

2010-11-12 Thread Hoekert, W.E.J.
Not here in The Netherlands. Van: histonet-boun...@lists.utsouthwestern.edu namens Laurie Colbert Verzonden: vr 12/11/2010 15:42 Aan: godsgal...@aol.com; Histonet@lists.utsouthwestern.edu Onderwerp: RE: [Histonet] Formalin Not us - in Pasadena CA. -Origina

RE: [Histonet] PMS2

2010-09-16 Thread Hoekert, W.E.J.
We used the same antibody but then at 1:1200, retrieved with PH9 (PT module buffer 4, thermo) on a labvision stainer. We had beautiful results. Now, we switched to Ventana Benchmark XT (1:50, 30' cc1 + amplification) and our results are quite poor for PMS2: many cytoplasmatic staining. So i

RE: [Histonet] Glut1 antibody

2010-06-23 Thread Hoekert, W.E.J.
We switched to Neomarkers (polyclonal) (RB 078-A). Willem Hoekert OLVG, Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens Martha Ward Verzonden: di 22-6-2010 22:10 Aan: histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] Glut1 antibody I

[Histonet] Smad4 on a Ventana Benchmark XT

2010-04-14 Thread Hoekert, W.E.J.
Hi Histonetters, We are thinking of taking the Ventana Benchmark XT, and I am testing it right now. So far, it is impossible to get the Smad4 up and running. I have tried everything: -Pretreatment with cc1 and cc2 for up to 90 minutes -Incubation of primairy AB for up to 90 minutes -Incubatio

[Histonet] How to do an EBER on cytological material?

2010-03-04 Thread Hoekert, W.E.J.
Dear histonetters, I was wondering if it is possible to do an EBER ISH on a cell-smear. What pretreatment should I use? 1: Fix the cells in formalin (let's say overnight, or else one or two hours), and than use the proteinase K digestion step. 2: Or maybe it is enough to fix the cells in cold

RE: [Histonet] HSV I & II

2010-03-02 Thread Hoekert, W.E.J.
Too bad that all these people did not respond to the entire group. We are using the one from Immunologic (polyclonal, ILP4114-C1). Is this one also not available anymore? Willem Hoekert OLVG, Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens

RE: [Histonet] [ SOLVED ][ Histonet ] Strange circles in IHC slides

2010-02-26 Thread Hoekert, W.E.J.
Are you sure that you don't introduce air bubbles when you put your slides into the coverplates? The antibody will not touch the tissue if there is an air bubble. Willem Hoekert Van: histonet-boun...@lists.utsouthwestern.edu namens Rene J Buesa Verzonden: do 2

RE: [Histonet] CISH - HER2

2010-01-06 Thread Hoekert, W.E.J.
We are using Spot Light her2 Cish from invitrogen (manual, FDA approved). Willem Hoekert Pathology OLVG, Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens Taylor, Jean Verzonden: di 5-1-2010 22:02 Aan: 'histonet@lists.utsouthwestern.edu'; 'ih..

RE: [Histonet] To my fellow histotechs....

2009-12-18 Thread Hoekert, W.E.J.
* Cyclin D1 : NeoMarkers, SP4 * CD56 : NeoMarkers, 123C3.D5 * CD138 : Dako, M115 * Kappa : Dako, A0191 (Polyclonal) * Lambda : Dako, A0193 (Polyclonal) * MUM-1 : Dako, MUM1p Willem Hoekert Pathology, OLVG The Netherlands Disclaimer: Dit e-mail bericht is uitslui

RE: [Histonet] CD68 on FFPE human tissue

2009-11-18 Thread Hoekert, W.E.J.
Yes, we too. Pretreatment with trypsin. Van: histonet-boun...@lists.utsouthwestern.edu namens Richard . Verzonden: wo 18-11-2009 15:09 Aan: Kim Merriam CC: Histonet Onderwerp: Re: [Histonet] CD68 on FFPE human tissue PG-M1 from dako 2009/11/18 Kim Merriam >

RE: [Histonet] Labvision immunostainers skip slides

2009-10-15 Thread Hoekert, W.E.J.
which contains tween already. And yes, we do get good support, but still the problems are not solved. Thanks again, Willem Hoekert Pathology Lab, OLVG Amsterdam The Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens Hoekert, W.E.J. Verzonden:

[Histonet] Labvision immunostainers skip slides

2009-10-09 Thread Hoekert, W.E.J.
Dear Histonetters, A few months ago, we bought 2 Labvision Autostainers 720 from Thermo (immunostainers). We have some bad experiences with them. They seem to skip some slides every now and than (it happens maybe once every 3-4 weeks, we stain about 150 - 200 slides a day). If so, our positiv