Re: [Histonet] HMGB and RAGE; TLR2, TLR4 antibodies
thanks Amos which one did you used ? you have catalog number ? On Fri, Oct 1, 2010 at 9:44 PM, Amos Brooks amosbro...@gmail.com wrote: Hi, I am currently working on HMGB1. It is a weird one! If you go by the data sheets you will get great nuclear staining on all the nuclei. This is not what you are after if you have necrotic tissue. What you want to see is the nuclei in necrotic tissue fade out and have the cytoplasm take it up. I am cutting the dilution to 1: 50 and 1:100 since we were just starting to see blushes of cytoplasmic staining and no nuclear label at higher dilutions. Usually you get staining that fades out at higher dilutions. This one was really odd. Good luck, Amos Message: 13 Date: Thu, 30 Sep 2010 04:55:02 +0200 From: Fabrice GANKAM gan...@googlemail.com Subject: [Histonet] HMGB and RAGE; TLR2, TLR4 antibodies To: histonet@lists.utsouthwestern.edu Message-ID: c4a46c631cd14a86aad3ce3abb29a...@pcdegankam Content-Type: text/plain; charset=us-ascii Hey guys Just wanted to know if any of you had some luck with an antibody aigainst rat HMGB1 and its receptors RAGE, TLR2, TLR4. W Thanks Dr Fabrice GANKAM UTSW ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] HMGB and RAGE; TLR2, TLR4 antibodies
Hey guys Just wanted to know if any of you had some luck with an antibody aigainst rat HMGB1 and its receptors RAGE, TLR2, TLR4. W Thanks Dr Fabrice GANKAM UTSW ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] multi-timer
Hey guys Was wondering if any of you ever used a phopho NFKB p65 antibodu and which one you prefer with what antigen retrieval method if applies. I'm working on FFPE rat tissue Thanks. Fabrice ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] L-DNAse II vs neutrophil (leukocyte) elastase inhibitor
Hi guys Would like to know if any of you have used the L DNAse II or the serpin B1 antibody in the rat. very few references mentioning this antibody. Sigma has one prestige antibody for serpin B1 but do no know if it will work on rats. I was wondering if the DNAse II is the equivalent of the L DNAse II (product of the degradation of serpin) thanks for your help ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: L-DNAse II vs neutrophil (leukocyte) elastase inhibitor
Hi guys Would like to know if any of you have used the L DNAse II or the serpin B1 antibody in the rat. very few references mentioning this antibody. Sigma has one prestige antibody for serpin B1 but do no know if it will work on rats. I was wondering if the DNAse II is the equivalent of the L DNAse II (product of the degradation of serpin) thanks for your help ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] anti 8 oxoguanine, 8 hydroxyguanine
Dear histonetters Will like to use the anti 8 hydroxyguanine (staining the oxidised DNA and RNA to asses for free radical damage to nucleic acids)for IHC but tried the antibody from abdserotec with dissapointing results. We tried it on liver slides but LOT of background. Could any one advise on other antibody or other protocol ? Thanks y'all Dr Fabrice GANKAM Intern ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] anti 8 hydroxyguanosine
Hey guys was wondering if any of you used the anti 8 hydroxyguanosine to detect free radical induced DNA and RNa damage. which antibody is the best. we tried the one from abdserotec and it is a disaster. the background is just horrible. any idea ? 2010/4/26 Perry, Margaret margaret.pe...@sdstate.edu In the Brown and Hopps gram stain can I substitute 37 g in 100 ml of powdered paraformaldyhyde for the 37% formalin or do I need to buffer it? ___ 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] iNOS/nitrotyrosine antibody
Dear All Looking for the best possible iNOS and nitrotyrosine antibodies that work with rat brain tissue on paraffin section I used the one from chemicon in the past but results were not so great I also use the one from labvision with disappointing results. Any help ? Thanks Dr Fabrice GANKAM -Message d'origine- De : histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] De la part de Bryan Llewellyn Envoyé : lundi 5 avril 2010 21:37 À : Histonet; malbena...@gmail.com Objet : Re: [Histonet] 72644.18148...@web05.mail.gq1.yahoo.com Comparing the situations in different countries can be very confusing. I traied in the UK (many years ago) and have lived in Canada for a long time, but I do have some (limited) information about the US system. First off, Medical Laboratory Technology in the UK and Canada includes histotechnology as one of the integral subject areas, but the US does not. Histotechnology is a standalone subject there, by and large. The ASCP is different from the Health Professional Council (used to be Council for professions supplementary to medicine when I lived there). That is a licensing body, and its function is carried out by some state agencies (in the US) and some provincial agencies (in Canada). However, not all states and provinces require licensing to work as a medical laboratory technologist/histotechnologist. The ASCP used to run the commonest US qualifying exams (still do ??, I am not sure) and kept a registry of qualified technologists, although there are others systems. In Canada it is done by the CSMLS (Canadian Society for Medical Laboratory Sciences). The equivalent organisation in the UK is the IBMS (Institute of Biomedical Sciences). In Canada it is possible to take specialty training in a subject area at both initial level and post initial level. So you can be an RT (Registered Technologist) in medical laboratory technology generally, or an RT in cytology or electron microscopy as examples. All RTs can take advanced examinations as general or specialist technologists, depending on their initial RT status. There used to be a third level (Fellowship in the CSMLS) but it was abandoned because so few technologists took it. That was about the same level as the UK three part exam. An applicable BSc is now required in Canada to advance post RT. As to 16 year olds in labs. I started work in the UK in Hackney six days before my 17th birthday in 1960. A month later I was doing haemoglobins by finger stick with Hagedorn needles, ESRs and going around the wards. A year later I was well versed in clinical chemistry (urea, glucose, bilirubin, alkaline phosphatases etc - all done manually with what passed for micro methods in those days. Students like me did about 80% of the work in those days because the profession was expanding so fast due to the introduction of the public health care system in Britain. Things change, and that just would not be allowed today. I suspect that 16 year olds doing grossing is very unusal and in the US would likely be viewed as an invitation for the pathologists to be sued. Remember, it was April 1st! In the US the CAP (College of American Pathologists) is involved in an accreditation system with other agencies. In Canada the CAP (Canadian Association of Pathologists) is a player in some accrediatation systems, but in Canada health care is legally a provincial responsibility, so accreditation is done by provincial agencies for each province. That is also the reason licensing varies from province to province here. Our country wide qualifying system by the CSMLS is a fortunate anomoly that nobody wants to change because it works so well for us. I hope this explains a little. Bryan Llewellyn - Original Message - From: Malika Benatti malbena...@googlemail.com To: Mark Tarango marktara...@gmail.com Cc: histonet@lists.utsouthwestern.edu; Andrew Burgeson nap...@siscom.net Sent: Monday, April 05, 2010 11:03 AM Subject: Re: [Histonet] 72644.18148...@web05.mail.gq1.yahoo.com I am very confuse reading every email reply to this tread also I would be really grateful if someone could enlighten with regard to what is the comment practice in the US. Having been trained as a histotechnologist although we are call Specialist Biomedical Scientist in the UK, we cannot practice unless we are fully registered with the Health Professional Council HPC, which I believe has the same role as the ASCP. Every 2 years we may be audited a demonstrate that we fully comply with HPC regulation and CPD or lose or registration. All laboratories are accredited by the Clinical Pathology Accreditation CPA under the international organization for standardization legislation (ISO 15189). Laboratory accreditation happen every 2 years cycle for which the laboratory has to comply with a set of standard. During inspection accessor review everything
[Histonet] Free radical detection and cell death IHC
Sorry to post this again but had problems posting messages lately. Here is my problem, We do not have cryostat in our facility and I will like to test free radical production in situ in rat brain tissue Some of the methods involves homogenate of whole part of brain and assay with component reacting with free radical that does not give you the geographical distribution of free radical. I'm therefore looking for a methods involving IHC or IF in paraffin section. I checked for hydroethidine but it seems like the fluorescence fades with paraffin processing. Has any one used hydroethidine with paraffin processing ? Does anyone know another method of free radical or oxidative stress detection on paraffin section ? We also wanted to asses cell death by some marker with the same tissue (paraffin processed) the caspase stain is faint and we will like to find a selective marker of cellular death or irreversible damage (apoptotic or non apoptotic) that could be used in IHC and IF. Has anyone ever used PI on rat tissue after paraffin processing ? Any other marker ? I heard about the hydroxyprobe but does it work on paraffin embedded tissue ? Thanks for your help guys Fabrice ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Deleting extraneous parts of posts when replying
Hi, Just wondering if anyone has use hydroethinide to detect the free radicals (ROS) in the CNS or anyother tissue of rats after 4% formallin fixation and parrafin embedding. I wanted to used the hydroethidine on paraffin sections. I wonder if the fluorescence is lost by fixation (12hrs in 4% PAF) and paraffin embedding plus deparaffination. All the papers I reviewed used hydroethidine on frozen section but our facility does not have vibratome or cryostat. Please help Please Help. Fabrice ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
