[Histonet] Anti-Human Hepatocyte antibody clone OCH1E5
Hey Histonetters, Has anyone noted negative staining with the anti-human hepatocyte antibody Clone OCH1E5 on hepatocytes surrounding the centrilobular vein? We are staining mouse liver samples and get great staining on about 80-90% of the hepatocytes in normal liver but then see very low to frank negative staining only in this region. Thanks! Julie Julie Randolph-Habecker, Ph.D. Director, Experimental Histopathology Shared Resources Fred Hutchinson Cancer Research Center 1100 Fairview Ave N, DE-360 Seattle WA 98109-1024 Tel: 206-667-6119 Fax: 206-667-6845 jhabe...@fhcrc.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Luciferase IHC!
Thanks for the help! We actually tried antigen retrieval in Holy water in a pressure cooker for 10 minutes. I even brought in an old priest and a young priest to perform the staining but no luck! Sorry, no vendor for the holy water - the old priest makes his own. I enjoyed the laugh! This project has been hell! Thanks, Julie Message: 21 Date: Wed, 2 Nov 2011 08:46:28 -0500 From: Beckham, Sharon s...@stowers.org Subject: RE: [Histonet] Luciferase IHC To: 'O'Donnell, Bill' billodonn...@catholichealth.net, Kim Donadio one_angel_sec...@yahoo.com, Randolph-Habecker, Julie jhabe...@fhcrc.org, histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Message-ID: 2c40e43d1f7a56408c4463fd245dddf98aee2...@exchmb-02.stowers-institute.org Content-Type: text/plain; charset=iso-8859-1 You know sometimes I wish these emails had a LIKE button on them. I found this quite funny and wanted to LIKE it. Too much facebook for me! -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of O'Donnell, Bill Sent: Wednesday, November 02, 2011 8:19 AM To: Kim Donadio; Randolph-Habecker, Julie; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Luciferase IHC Actually, I have found that holy water reduces its sensitivity, greatly diminishes its signal and will destroy its reactivity. (Listen closely and you will hear the slide scream and growl and tell you no one likes you) (It's only Wednesday, what'll Friday look like?) -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kim Donadio Sent: Tuesday, November 01, 2011 7:01 PM To: Randolph-Habecker, Julie; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Luciferase IHC You have to soak it in holy water, then all H311 will break out Sorry, I couldnt resist. From: Randolph-Habecker, Julie jhabe...@fhcrc.org To: histonet@lists.utsouthwestern.edu Sent: Tuesday, November 1, 2011 6:49 PM Subject: [Histonet] Luciferase IHC Has anyone had good results staining for luciferase in FFPE tissue? If so, what antibody did you use. Thanks!! Julie Julie Randolph-Habecker, Ph.D. Director, Experimental Histopathology Shared Resources Fred Hutchinson Cancer Research Center 1100 Fairview Ave N, DE-360 Seattle WA 98109-1024 Tel: 206-667-6119 Fax: 206-667-6845 jhabe...@fhcrc.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 96, Issue 3 *** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Luciferase IHC
Has anyone had good results staining for luciferase in FFPE tissue? If so, what antibody did you use. Thanks!! Julie Julie Randolph-Habecker, Ph.D. Director, Experimental Histopathology Shared Resources Fred Hutchinson Cancer Research Center 1100 Fairview Ave N, DE-360 Seattle WA 98109-1024 Tel: 206-667-6119 Fax: 206-667-6845 jhabe...@fhcrc.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Turnaround time for Research Pathology
Folks, I am looking for some standard turnaround times for histology services including processing to embedding, processing to HE, and processing to HE and IHC. Does anyone have that data? Also, how do other folks handle standard turnaround time for research samples? We may get anything from 5 specimens to 300 specimens that just need paraffin processing and 1 HE. Or we may get a request for 10 samples for paraffin-processing, 10 slides each with 2 HE and 7 different IHC stains per case. Any insight on this would be very helpful. Thanks, Julie Julie Randolph-Habecker, Ph.D. Director, Experimental Histopathology Shared Resources Fred Hutchinson Cancer Research Center 1100 Fairview Ave N, DE-360 Seattle WA 98109-1024 Tel: 206-667-6119 Fax: 206-667-6845 jhabe...@fhcrc.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Control slides for Flag-Tag IHC
Folks, I am looking for a control for some Flag-tag IHC I am doing on FFPE tissue. I was wondering if someone could share some slides off of a block of flag transfected cells. THANKS!! Julie Julie Randolph-Habecker, Ph.D. Staff Scientist - Director Experimental Histopathology Shared Resource Fred Hutchinson Cancer Research Center 1100 Fairview Ave, N. DE-360 (Please note new location) Seattle WA 98109-1024 206-667-6119 jhabe...@fhcrc.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] CD4 and CD8 in FFPE mouse tissue
Folks, It's my annual inquiry as to whether someone has optimized immunohistochemistry for CD4 and CD8 on formalin-fixed paraffin mouse tissue. Over the years we have tried lots of different antibodies but I am wondering if there is anything new that we have not tried. Any leads would be great! Thanks, Julie Julie Randolph-Habecker, Ph.D. Staff Scientist - Director Experimental Histopathology Shared Resource Fred Hutchinson Cancer Research Center 1100 Fairview Ave, N. DE-360 (Please note new location) Seattle WA 98109-1024 206-667-6119 jhabe...@fhcrc.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Problems with mouse brains
Folks, I need some input on a problem we're seeing with some mouse brain samples. The samples are from new born mice P0 to P14 which have been fixed in 10% NBF (Fisher brand and well within expiration date) for 72 hours. They are then transferred to 70% etoh and processed on an 8 hour process. After the tissue is embedded, we are taking sagital sections, drying them overnight, and then baking overnight at 60C. They are then stained with HE. This has been working great but now all of a sudden we have very light HE staining, nuclear bubbling, and extensive tissue cracking. I also have noticed some formalin pigment. My first thought is that there was difference in formalin or time in formalin but that is not the case. I also wondered if the tissue might be exposed to excessive heat. We checked all of the temperatures in our processor, embedding center, and water baths - all were within tolerance. Any ideas what might cause all three artifacts? Could it be from inadequate paraffin infiltration? Any help would be greatly appreciated! Thanks, Julie Julie Randolph-Habecker, Ph.D. Staff Scientist - Director Experimental Histopathology Shared Resource Fred Hutchinson Cancer Research Center 1100 Fairview Ave, N. DE-360 (Please note new location) Seattle WA 98109-1024 206-667-6119 jhabe...@fhcrc.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet