Re: [Histonet] CD31 IHC on bone

Human or mouse? Are they trying to stain bone vasculature? If so I can recommend superior markers. Andrea > On Dec 12, 2014, at 10:06 AM, Connolly, Brett M > wrote: > > Hi all, > > A colleague is having problems staining CD31 on bone sections and is looking > for recommendations for specifi

Re: [Histonet] CD4 and CD8 background in mouse tumors frozen sections, help

Were these samples treated with an antibody based therapeutic and your secondary is detecting the therapeutic? If that's the case and they were treated with a human IgG biologic you will need to use a secondary cross adsorbed against human IgG. Andrea > On Dec 8, 2013, at 4:41 PM, "Erickson, J

[Histonet] Back processing Histogel

Has anyone back processed something that was both Histogel and FFPE embedded? If so, any hints or problems? Andrea ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] CD31 clone WM59

Does anyone know if this clone WM59 crosses to mouse CD31? Andrea ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Hight throughput IHC

Is anyone doing any type of high throughput IHC testing? If so I would love to pick your brain. Andrea T. Hooper, PhD andreahoo...@rocketmail.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listin

[Histonet] TMA & tissue sources

> (1) Do those of you who use cyno tissue have a favorite source for purchasing > FFPE and frozen blocks? (2) What is evetyone's favorite vendor for frozen TMAs? Thanks, Andrea ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.ut

[Histonet] Signal amplification

What kits/vendors are people using for their CSA/tyramide amplification protocols? Thanks in advance! Andrea On Jan 25, 2012, at 3:45 PM, "Mayer,Toysha N" wrote: > Also, to help cut the tissues, I face at room temperature (very slowly), then > I place my uterus/prostate blocks on ice with lo

Re: [Histonet] Nail Polish sealant

Amen Gayle, as always you hit the nail on the head. I never use nail polish sealant because of being burned so many times due to the isopropanol leaching into - and ruining - the sample. In addition if it gets onto pricey fluorescence lenses it's difficult to clean. It's not worth the risk, esp

[Histonet] F4/80

Has anyone stained NOD/SCID lungs for F4/80 or any macrophage marker? What is the expected staining pattern? Andrea ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Decal retrieving

Does anyone have experience with epitopes that are "retrieved" after decal? We have used three separate buffers (HCl, formic acid, formic acid/EDTA) and all retrieve. This is with three different proteins we are staining for, and we were curious if it is a common occurrence. Andrea ___

Re: [Histonet] Isotype specific polymers

Not sure, but perhaps you can buy a rabbit anti mouse IgG2a link antibody for example and then use a rabbit polymer. Andrea On Oct 5, 2011, at 4:19 PM, "Milne, Katy" wrote: > Hi everyone, > > I'm curious if there is such a thing as isotype specific polymers (ie will > react with mouse IgG2a

Re: [Histonet] Thick cryosections

Sorry for the late response. I have done immunofluorescence on 50 um cryosections of mouse host tumors and decalcified bone on super frost Plus slides. I melt them to the slide using body heat then dry them at RT for a few hours before storing at -80 deg C. They do not come off the slide. What

Re: [Histonet] ? on frozen tissue artifacts

Sounds to me Definitely more likely to be the freezing process. Do you know their process? Andrea On Oct 22, 2011, at 5:05 PM, histopath...@gmail.com wrote: > I never said these were muscles. These are tumors removed and frozen the day > before and sent to us on dry ice. We know not to use fro

Re: [Histonet] Problems with Frozen tissues

you can share with me? Andrea Hooper Sent from my iPhone On Nov 1, 2011, at 7:53 PM, Kim Donadio wrote: > may I ask why you are not fixing them? Even the stain the 1st gentlman was > doing(Beta Gal) recomended to have a formalin fixation apllied first. > > There are many ways to

[Histonet] RE: Staining with two primary antibodies from same host

I routinely stain with two primaries from same species. Mostly rat antibodies on mouse tissue. I have been fortunate enough so far not to have to do it in GFP containing tissue so I have utilized either commercially available FITc labeled primaries, or we label them ourselves. Then I use an ant

[Histonet] Double HRP immunostains

I do double HRP immunostains. Works well using DAB/AEC and True Blue ... but what are your favorite chromagens to use? Would love some feedback for inspiration. Andrea -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsout

[Histonet] CD115/c-fms/CSF-1R

Has anyone stained CD115 (aka c-fms/CSF-1R) in mouse tissue? Any suggestions of antibodies and protocols? Thanks, Andrea -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

histonet@lists.utsouthwestern.edu

POLL: What is everyone's favorite hematoxylin for work-horse H&E stains? What about eosin? I am re-evaluating the quality of our current set-up and think we can improve - especially for the frozen sections. Any suggestions/protocols/brand suggestions? Thanks, Andrea --

[Histonet] MCP-1/3

Anyone have any good antibodies to recommend for MCP-1 and MCP-3? THANKS! -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] mouse CD115

Anyone working with CD115 in mouse? If so, may I contact you as I have a few questions? Andrea Hooper anh2006 @ med.cornell.edu -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] mouse CD11c

Is anyone working on CD11c in murine tissues? If so, can we exchange emails? ... I have some questions. Andrea Hooper anh2...@med.cornell.edu -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman

RE: [Histonet] Embedding Stamp???

Sakura used to sell items called "Tissue Tampers" just for this purpose in different sizes. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Yeonju Shim Sent: Tuesday, April 14, 2009 8:54 AM To: histo

[Histonet] bovine vs donkey anti-goat IgG

Has anyone compared the bovine anti-goat IgG to the donkey anti-goat IgG secondaries from Jackson IR and seen whether there was a considerable benefit to using the bovine series? My BSA is gamma globulin free so it shouldn't make a huge difference for me, but I am just wondering . AND

[Histonet] microtome

What is the best microtome on the market these days? We are a research lab sectioning mainly mouse tissue, and mainly decalcified bone. We have beginners to experts in the lab all who will need to use it. It's been a while since I have been in the market for a microtome, so although I have m

RE: [Histonet] list of special stains

DAKO has (or had as it's been some years since I got mine) a FREE Guide to Special Stains introductory manual, which was great for such info. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Zerfas, Patr

[Histonet] DyLight fluorophores

Any thoughts on the new DyLight series of fluorophores from Jackson ImmunoResearch? http://www.jacksonimmuno.com/technical/DyLight.asp -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histo

Re: [Histonet] Histology for Zebrafish

Have you checked out the ton of protocols (including histo) available at ZFIN online: http://zfin.org/zf_info/zfbook/zfbk.html Hi, I need to work on zebrafish for the first time and would like to get some general protocols for fixation and IHC staining. My investigator used 4% PFA for 2 hrs

Re: [Histonet] staining brain vessels

Hi Colleen, Although I agree that you have to make sure your markers are good for the particular species you work with, I respectfully disagree that CD34 does not "work" in mouse. CD34 is one of the best vascular markers for mouse paraffin sections, of course it is absolutely tissue specif

Re: [Histonet] CD31 on FFPE mouse tissue

Does it have to be CD31? I have a bunch of good protocols for mouse vessels but each tissue requires a different antibody. CD31 in FFPE is never reliable in my experience. Which tissue are you using? Andrea Kim Merriam Sent by: histonet-boun...@lists.utsouthwestern.edu 02/13/2009 12:27

[Histonet] Leica Thermal Block

Does anyone use the Leica Thermal Block? How does it work? Do you need to prewarm it? I was told to leave it in the cryostat but that doesn't work as it gets as cold as the chucks ... ? Thanks, Andrea -- ___ Histonet mailing list Histonet@lists.utsou

[Histonet] EphB4 & ephrinB2

Is anyone staining these proteins successfully in mouse tissue? If so, where are your antibodies from? Thanks. -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Autoflourescence

10-15 um seems extremely thick for paraffin sections. Can you get thinner sections? Say no thicker than 6um? Unfortunately autofluorescence is a fact of life with FFPE sections, hence why people normally do not choose to do immunofluorescence on FFPE sections. Are you tethered to using immunofluo

Re: [Histonet] staining rat tissue with monoclonal antibody?

Just make sure you use an anti-mouse IgG antibody that is not cross reactive to rat IgG. Jackson ImmunoResearch is the best place to find these reagents ... I suggest the donkey as host, it works best for me, least background. Beautiful results ... Good afternoon. I just had someone ask me ab

[Histonet] murine BM monocytes

Is anyone doing F4/80 on frozen mouse bone sections? How about other monocyte markers? Would love to hear about any experiences. Thanks, Andrea -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listi

[Histonet] Cryostat safety question

The discussion on microtome safety begs me to ask a cryostat question We have a Leica CM3050 cryostat and love it! How are people (and perhaps only those in research do this) removing their tissue from the chucks for future use? We often just section a few slides worth then put the block

Re: Re: [Histonet] Capillary counting

What animal? What organ? In frozen or paraffin? So any one can provide some cell makers for staining of blood vessels? I only know CD31. 2008-12-25 TF -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.

[Histonet] mouse dendritic cells

Is anyone staining for mouse dendritic cells either in paraffin or frozen sections? If so, what markers/antibodies are you using? Thanks, ANDREA -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/lis

RE: [Histonet] IHC on paraformaldehyde-fixed

Good point. I never tested it directly side-by-side to check for the effects of 1% methanol directly. It could be old-fashioned paranoia. However, it was a variable I wanted to omit as I know methanol (we are talking 100%) has deleterious effects to some mouse antigens. Therefore, it wasn't som

[Histonet] NG2

Is anyone using NG2 on FFPE sections of mouse tissue? If so, I could use some details of your protocol and antibody. Thanks!! Andrea -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histon

Re: [Histonet] IHC on fresh frozen

Try fixing for 5-10 min in PFA. At 10:16 AM -0500 12/3/08, FU,DONGTAO wrote: Hi, all Recently I did some IHC(chromagen methods) on mouse fresh frozen tissues, mainly using insulin antibody on pancreas. The image is much fuzzier compare to paraffin embedding tissue. And the staining also sm

[Histonet] Testes vascular markers

What are the best markers to stain testicular endothelium? Thanks, Andrea -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

RE: [Histonet] Presidential Voting Infomation

Honestly who cares. Let's just report to Listserv manager and move on. -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] double trouble

You cannot do what Paula recommends below without extra blocking steps, if the primaries are raised in the same species as the second secondary is raised. This is a rough outline of what I recommend: 1st primary antibody (X-anti-Y) 1st biotinylated secondary antibody (Fab anti-X) Streptavidin-H

[Histonet] mouse heart endothelium

Is anyone routinely staining for mouse heart ECs in FFPE sections? If so, what markers and protocol are you using? Thanks, ANDREA -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] X-Gal stain of bone

Dear Markus, We do X-Gal staining on bone and in the best scenario the animals are perfused with glutaraldehyde (GA). PFA tends to quench the beta-gal activity and the results may not be 100% accurate (underestimation of positivity). Though if you want to do IHC on these samples, I am sure y

Re: [Histonet] Fw: platelets and IHC

Can you post a pic of this non specific staining? What organ is it? Is it inside blood vessels? At 5:27 PM -0500 10/7/08, Jan Shivers wrote: Addendum... I should have included this information in my original posting... I'm already doing a 3% H2O2 peroxidase quench (10-15 minutes) and a 10%

Re: [Histonet] ever heard of these antibodies?

Anti-cleaved caspase 3 works beautifully from Cell Signaling. www.cellsignal.com For IHC: 1.Nephrin 2.m-TOR 3.caspases Have you ever used them on FFPE? did it work ? where did you but them? Thank you Kimberly C. Tuttle HT (ASCP) Pathology Biorepository and Research Core University of Mar