Re: [Histonet] Coverslipper

Happy Thursday, We would like to sell our slightly used but in excellent condition Dako Coverslipper which was purchased in 2010 (purchase price was $ 25K). The instrument can handle up to 600 slides per hour making it one of the fastest on the market. In addition to the flexibility of the

[Histonet] X-gal staining

Hello histonetters, I have request from a scientist to perform X-gal staining on the formalin fixed paraffin-embedded kidney. I was wondering if there is any possibility to do this staining on FFPE tissue as well as if there any possibility to perform a peroxidase DAB staining for the X-gal?

[Histonet] job search in Chicago area

qualifications and accomplishments. I am very interested in meeting to discuss how I can best contribute to the work in research laboratory. Sincerely, Naira Margaryan Naira V. Margaryan, D.V.M., Ph.D. Senior Research Scientist West Virginia University Robert C. Byrd Health Sciences Center Department

Re: [Histonet] Tissue scrolling

I do the same way! Good luck! Naira From: Colleen Forster Sent: Sunday, September 8, 2019 12:43 PM To: John Garratt Cc: Erin McCarthy ; histonet-request Subject: Re: [Histonet] Tissue scrolling We cut alot if these. This is the protocol: 1. The microtome,

Re: [Histonet] IHC mouse optomization

Biocare Medical has pure ready to use secondary.. Naira -Original Message- From: Blanca Lopez Sent: Wednesday, June 12, 2019 10:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC mouse optomization Morning, I need a good advice in optimizing Mouse antibodies for

Re: [Histonet] Coverslipper

Happy Thursday, We would like to sell our slightly used but in excellent condition Dako Coverslipper which was purchased in 2010 just for $10K (purchase price was $ 25K). The instrument can handle up to 600 slides per hour making it one of the fastest on the market. In addition to the

Re: [Histonet] Coverslipper

Happy Friday, We would like to sell our slightly used but in excellent condition Dako Coverslipper which was purchased in 2010 $10K (purchase price was $ 25K). The instrument can handle up to 600 slides per hour making it one of the fastest on the market. In addition to the flexibility of the

Re: [Histonet] Coverslipper

Happy Thursday, We would like to sell our slightly used but in excellent condition Dako Coverslipper which was purchased in 2010 $10K (purchase price was $ 25K). The instrument can handle up to 600 slides per hour making it one of the fastest on the market. In addition to the flexibility of

[Histonet] Polyclonal Rabbit Anti-Cytokeratin

Hello, I was asked to do IHC using the Polyclonal Rabbit Anti-Cytokeratin, Wide Spectrum Screening, Polyclonal, Unconjugated, Ig fraction, (Z062201-2). Ca anyone help me with AR to use and concentration to start. Thanks in advance, Naira ___

Re: [Histonet] Coverslipper

Happy Thursday, We would like to sell our slightly used but in excellent condition Dako Coverslipper which was purchased in 2010 $10K (purchase price was $ 25K). The instrument can handle up to 600 slides per hour making it one of the fastest on the market. In addition to the flexibility of

[Histonet] coverslipper CR10030

Dear histinetters: We can offer our slightly used DAKO coverslipper CR10030 for purchase. Let us know if you are interested. Thanks, Naira Ranked nationally in all 10 pediatric specialties by U.S. News & World Report (LCHOC Ver 1.0) This message contains confidential information and is

[Histonet] PD-L1 22c3 for IHC (paraffin)

Dear Friends, Can any of you share with me a full detailed protocol (time and dilutions) after deparaffinization for the PD-L1 22c3 for IHC (paraffin) to use on DAKO (not Link 48) autostainer?

[Histonet] PD-L1 and TGFb for FFPE human tissue

Good morning everyone, I was wondering if anyone can give me an advise regarding the best Abs for the PD-L1 and TGFb for FFPE human tissue (except DAKO Abs, please). Thanks in advance, Naira Ranked nationally in all 10 pediatric specialties by U.S. News & World Report (LCHOC Ver 1.0)

[Histonet] Immunoperoxidase Protocol on Cytospin

Dear histonetters: I never done ICC. A scientist brought me a slide with Cytospined cells on for IHC. I am going to fix with 10%NBF (it is only what I have now) then wash with TBST. May I skip AR step? May I use my usual IHC protocol and reagents which I usually use for FFPE tissue: blocks

Re: [Histonet] Recycled reagents in VIP processor

Absolutely agreed! Naira -Original Message- From: Frazier, John [mailto:john.fraz...@roche.com] Sent: Tuesday, February 28, 2017 2:31 PM To: Jay Lundgren <jaylundg...@gmail.com> Cc: Margaryan, Naira <nmargar...@luriechildrens.org>; Gareth Davis <garethdavisy...@gmail

Re: [Histonet] Recycled reagents in VIP processor

WOULD LIKE TO KNOW AS WELL - IT IS WASTE OF MONEY Thanks, Naira Ranked nationally in all 10 pediatric specialties by U.S. News & World Report (LCHOC Ver 1.0) This message contains confidential information and is intended only for the individual named. If you are not the named

[Histonet] rat vomeronasal organ (VNO) fix, decal, proc. and paraffin embedding protocol

Dear Histonet Colleagues, I hope you had very nice and warm holiday season . We are interested in a routinely used protocol to decal (if needed), process and paraffin embed the rat vomeronasal organ (VNO). It would be greatly appreciated if anyone would be willing to share a protocol for

Re: [Histonet] Ki67 antibody

Me too, love it! Naira Ranked nationally in all 10 pediatric specialties by U.S. News World Report (LCHOC Ver 1.0) This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or

RE: [Histonet] Re: squamous cell contamination on slides

Hi Hestonetters, I need to do IHC of SC4MOL on FFPE sections. If any of you have been done this staining before, please send me your procedure. Thanks in advance, Naira Ranked nationally in all 10 pediatric specialties by U.S. News World Report (LCHOC Ver 1.0) This message contains

RE: [Histonet] Histology as art!

Hi Emily, I don't mind if it is healthy tissue! It shows that even small cell or huge artery =all from the mother nature, don’t you think? Naira Ranked nationally in all 10 pediatric specialties by U.S. News World Report (LCHOC Ver 1.0) This message contains confidential information

[Histonet] FW: Hemoglobin

Hi histonetters, Do you know how we can stain for hemoglobin in FFPE zebrafish tissue? Thanks in advance, Simone. Ranked nationally in all 10 pediatric specialties by U.S. News World Report (LCHOC Ver 1.0) This message contains confidential information and is intended only for the

RE: [Histonet] Mouse lung IHC false positive/background issue

I have same problem for other Abs using in on mouse lung tissues... My secondary and tertiary have no problems on other tissues for negative controls as well Naira ** Confidentiality Note: This message (including any attachments) is intended for the use of the named

[Histonet] IHC (DAB) after HE?

Dear Histonetters, I would like to perform IHC (DAB) after HE. Do I need to remove HE? If answer is YES, my question HOW? Can I ask the protocol for that? Thanks in advance, Naira ___ Histonet mailing list Histonet@lists.utsouthwestern.edu

[Histonet] zebrafish and IHC

Dear Histonetters, I am trying to do IHC on FFPE 4 weeks old zebrafish with different Abs. Is there a trick working with zebrafish? I am using the same IHC protocol I always use on human and mouse tissue and my Abs are suppose to work on fish as well as human. I run human and fish sections

[Histonet] HE reagents

What companies HE reagents (catalog preferably) are the best for simple HE staining? Thanks in advance, Naira ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] proliferation and apoptosis

Hi histonetters, I am looking for the good markers to detect (separately) proliferation and apoptosis of cells in tumor sections. Unfortunately, KI-67 stains apoptotic bodies as well as proliferated cells; and the tunnel assay shows both apoptotic body and proliferation. Any suggestions for

[Histonet] reagents without expiration dates

My oldest was from 1965, I can't bit Rene:) Naira -- Message: 12 Date: Thu, 2 Feb 2012 13:14:53 -0800 (PST) From: Rene J Buesa rjbu...@yahoo.com Subject: Re: [Histonet] reagents without expiration dates Ha.Ha,Ha!!! I used to prepare staining solutions with

[Histonet] emergency question-IHC interruption

Hi, Where I can stop my IHC, in what step? I started deparaffinization, but I need to stop and run my IHC tomorrow. Can I leave my slides in 95% Ethanol, or Water or I have to do HIER? Where to store slides: in refrigerator or room. Naira ___

FW: [Histonet] emergency question-IHC interruption

Thanks to all who are kindly and quick answered on my question. Naira On Thu, Oct 13, 2011 at 4:20 PM, Margaryan, Naira nmargar...@childrensmemorial.orgmailto:nmargar...@childrensmemorial.org wrote: Hi, Where I can stop my IHC, in what step? I started deparaffinization, but I need to stop

[Histonet] HE on cells, protocol

Hi Histonetters, I have to do HE staining on cells that were growing up on coverslips. Can anyone send me a good protocol to fix these cells on same coverslip and HE staining, please? Thanks in advance, Naira ___ Histonet mailing list

[Histonet] melanin counterstaining

Good morning, I have heavily pigmented melanocytic cells and trying to differentiate between melanin pigment and chromogen (DAB) by using Azure B. After IHC with DAB I incubate slides for 30-45 min in the working solution of: * 4ml of a 25mg/ml aqueous solution of Azure B * 3.4ml of 0.1M

[Histonet] Retirement

I am 15-18 years out from retirement also. Count me in as well Naira ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] How do you call...

I totally agree with Rene J.: there is anatomy and physiology becomes pathological that is why need to call Pathological Anatomy and Pathological Physiology Foreign people learn what they hear usually that is why Americans have to watch there language when speaking and use only correct words!

[Histonet] FW: How to remove DAB to restain with DAB

Hello, I would like to repeat my DAB staining on the same slide with DAB I run before. I know that acid alcohol will remove hematoxylin but how to remove DAB? I appreciate to any suggestion. Thanks in advance, Naira ___ Histonet mailing list

[Histonet] kits for LCM archival FFPE tissue

Hi all, I am about to perform PCR on archival FFPE tissue after LCM. My LCM system is from Arcturus and my tissue is on membraned glass slides. My questions are: 1. What Kit to use to get the highest yields and quality RNA? 2. Can anyone who has been doing it for the past 2-3 years suggest

[Histonet] TGFβ RII

Hi histonetters, I need suggestion and help with the TGFβ RII (D-2): sc-17799 Ab, which should show a cytoplasmic staining. My problem is that I am getting a beautiful nuclear staining only. How I can fix this problem? What to pay attention on and what to change in the usual protocol? My AR is

RE: [Histonet] ihc slides

Hi Cynthia, I agree with Greg, you have to re-do your staining from beginning (from AR step) after removing the hematoxilin. Naira ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Antigen Retrieval for 10u sections in IHC

Hi tistonetters, I have to do IHC on 10µ sections. Is procedure for Antigen Retrieval same like for 4-5µ (time and temperature) Thanks in advance, Naira ___ Histonet mailing list Histonet@lists.utsouthwestern.edu

[Histonet] RE:Tap Water or filtration system

What if we suggest to use ddH2O both for TBST preparation and in autostainers (before and after IHC) because the PH level is vary in different states. Naira ** Confidentiality Note: This message (including any attachments) is intended for the use of the named recipient(s) only

[Histonet] FW: How to remove Hematoxilin

Hi histoworld, I would like to repeat my staining on the slides already coverslipped but need to remove hematoxilin first. How to remove hematoxilin? Is it need to repeat Antigen retrieval? Thanks in advance, Naira ___ Histonet mailing list

[Histonet] RE: coverslipper

Thanks a lot to all of you answered me. I was surprise nobody mentioned coverslipper from DAKO. Are any of you have any experience with DAKO's coverslipper? Again Thanks to all, Naira Subject: coverslipper Hi Colleges, I need your opinion about coverslip by hand vs. using machine. If you

RE: [Histonet] Incubation of Ab more then 1 hour in autostainer

-Original Message- From: Liz Chlipala [mailto:l...@premierlab.com] Sent: Thursday, March 18, 2010 3:05 PM To: Margaryan, Naira; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Incubation of Ab more then 1 hour in autostainer We have done up to an hour but not longer, if I were you

[Histonet] Incubation of Ab more then 1 hour in autostainer

Hi Histonetters! I have a stupid question but I Have to ask. Does anyone perform an Incubation of Ab that required more then 1 hour (2-3 hours) in autostainer? Does autostainer keep slides wet or slides sometimes are getting dry? I know that slides should not be dry in any step of IHC.

[Histonet] Bleach in autostainer cost problems?

Hi Histonetters, Have you ever had the problem I am having now: light or totally negative staining after service and cleaning the machine with bleach? To explain it better: I have immuno-autostainer from TermoFisher that was working well before I had service from the company. After a

[Histonet] IF staining on FFPE tissue

Dear Histonetters, Hopefully you are enjoying a great Holidays with your friends and family! I have been asked to perform . I got positive and negative absolutely identical with beautiful nuclear DAPI and very red RBC. What to do or what to change in my protocol to avoid RBC and background?

[Histonet] RE: IF staining on FFPE tissue

Dear Histonetters, Hopefully you are enjoying a great Holidays with your friends and family! I have been asked to perform . I got positive and negative absolutely identical with beautiful nuclear DAPI and very red RBC. What to do or what to change in my protocol to avoid RBC and background?

[Histonet] anti-human Notch4

Hi friends, My PI is asking for Notch 4 IHC on FFPE tissue. Can any of you suggest me a good Ab (preferably mot mouse but anti-human Notch4) and protocol please! Thanks in advance, Naira Naira V. Margaryan, D.V.M., Ph.D. Research Scientist Children's Memorial Research Center 2300

[Histonet] AlkPhos

Dear histonetters, I have to order AlkPhos kit to my IHC and, because I did not use it for the past 5-6 years, I do not know what company will best to order from. Any suggestions are appreciated, Naira ___ Histonet mailing list

[Histonet] human macrophages in human melanoma xenografts

Hi Histonetters, I would like to be able to look at human macrophages in human melanoma xenografts raised in mouse. Could you please suggest me a best Ab for ICH and protocol? Thanks in advance, Naira ___ Histonet mailing list

RE: [Histonet] human macrophages in human melanoma xenografts

; Margaryan, Naira; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] human macrophages in human melanoma xenografts F4/80 is a mouse macrophage marker. If she wants to detect human macrophages in a mouse background she will need to use a mouse anti-human CD68. She will need to run

[Histonet] Isotype background

Hi Adam, I always do the isotype control parallel with a no-primary control in parallel with my real Ab. May I suggest you to use Protein block serum free and pure Ab diluent without adding 2% donkey serum? Try this and let me know your results. If histonetters think I am wrong, fill free

[Histonet] black DAB instead of brown

Dear Histometters: I am getting black staining with my DAB instead of brown. I use usual protocol with AR-pH6, H2O2, Avidin/Biotin, PB, 1º, biotinylated 2-dary, streptavidin then DAB. Does any of use know this kind of problem? and What exactly can coast this artifact? Thanks in advance, Naira

[Histonet] acid phosphatase in tissues in routine IHC

Hi histonetters, Around a month ago, I asked a question and did not get any answer, but I really would like to have any thoughts from you about the: Would the presence of acid phosphatase in tissues cause non-specific background staining when doing routine immunoperoxidase staining -- DAB? If

[Histonet] type of paraffin and polymer

Dear Histonetters, I quite need answer as soon as it possible, PLEASE! I am working with mouse tissue and tumors. What type of paraffin is the best for processing and embedding to cut 4µ nice sections? Thanks in advance, Naira ___ Histonet mailing

[Histonet] RE: reagents for IHC

Hi Everyone, I do appreciate for all suggestions I get form most of you! Have a nice week, Naira ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] reagents for IHC

Hi everyone, How are you experiencing the economic pressures and price changes for REAGENTS? I am sorry, but I just bought reagents from DAKO and, for the price I paid for 125 ml before, I got 15-50 ml :( I am ready to switch to another company, but I need your suggestion about reagents for

[Histonet] bone fixation and calcification

Dear Histonetters, I am new in fixation and processing bones. I need your full protocol with details how to fix and process mice bone to visualize the tumor metastases? Thanks in advance, Naira ___ Histonet mailing list

[Histonet] Fixation of Formalin vs Bouin's for IHC

Hi histonetters, Please help. What is difference in procedure for IHC between 10%Formalin fixed tissue and in Bouin's fixed tissue? I used to use 10%FFPE tissue with Citrate buffer Antigen Retrieval for my IHC. Do I have to change my protocol for the Bouin's fixed tissue? Hope for you soon

[Histonet] Trypsin versus other AR-s

Hi Dears, It is Friday, but not a weekend yet:-):-( I just got a request from my PI to figure out: 1. How often now day's people use Trypsin (EDTA, Proteinase K or E) as an Antigen Retrieval for FFPE. 2. Why or is the Citrate Buffer pH6 more usable??? 3. Is Trypsin very

[Histonet] keratin 18

Hi histoneters, Please help me to find an Ab to detect keratin 18 in mouse tissue, this means that at has to be not mouse Ab. Thanks in advance, Naira ___ Histonet mailing list Histonet@lists.utsouthwestern.edu

[Histonet] chicks' embryos and LCM

Dear histoneters, I am going to start the LCM work on chicks' embryos and now more questions are coming up :-) 1.What kind of brand of blades will you suggest to use for crysectioning (Catalog ## will be very good)? 2.After placing glass slides in LCM what kind of caps will you

[Histonet] RE: HE help (Weems, Joyce)

Hi Roger, I usually use Bluing reagent (Richard Allan Scientific, part of Fisher) after Hematoxylin and tap water, then wash again before eosin. Try it and you will see big difference. I just love it! All the best, Naira --