[Histonet] Movat Pentachrome - Need to enhance Alcian Blue
Hi All, I had wonderful help from this site for my Woodstain Scarlet-Acid Fuchsin binding collagen problem. Now that this is fixed I have realized that the Alcian Blue is not staining my excessively fixed samples (Human Aorta & Aortic Valve, formalin fixed 2-4 years, embedded in paraffin and stored at RT for ~6 years). I have some samples fixed for only 2-6 months and these are fine but the samples fixed for 2-4 years do not uptake the Alcian Blue. I have taken it up to 1 hr in the 1% Alcian Blue solution followed by 10 min in 56*C Alkaline Alcohol solution. I do a 1hr Bouin's 50*C to start (I have also tried Bouin's at 60*C). Any ideas on how I can get the Alcian Blue to stain these excessively fixed samples or possibly a substitute? I need to see the proteoglycans/GAGs. Thank you Krista ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Renee Grow is out of the office.
I will be out of the office starting 05/27/2013 and will not return until 06/06/2013. I will respond to your message when I return. This communication may contain protected health information (PHI) that is legally protected from inappropriate disclosure by the Privacy Standards of the Health Insurance Portability and Accountability Act (HIPAA) and relevant Tennessee Laws. If you are not the intended recipient, please note that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this message in error, you should notify the sender immediately by telephone or by return e-mail and delete this message from your computer. Direct questions to the Blount Memorial Hospital Privacy Officer at 865-977-4675. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] MDM2 on lipsarcoma IHC
Hi Gudrun, We only do FISH for MDM2 but it is positive for gene amplification in both well differentiated liposarcomas and dedifferentiated liposarcomas. Lipomas and other sarcomas should be negative as far as I've seen. I haven't worked with the antibody or seen that staining, but I'd guess that you are right and the protocol needs to be brought down. That link you have in your e-mail seems to imply that the antibody stains nearly all nuclei though. Maybe the antibody isn't just staining the overexpressed cells... but all of them? "Ubiquitous nuclear expression" doesn't sound very specific for the cells that are positive for gene amplification. Mark On Mon, May 27, 2013 at 8:37 AM, Gudrun Lang wrote: > Hi all! > > We try to improve our MDM2-staining and purchased a new antibody. This one > stains all nuclei in liposarcoma but also in normal surrounding tissue and > lipoma positiv. > > On the Protein-Atlas-site MDM2 is described with "Ubiquitous nuclear > expression". On the other hand I read, that well differentiated liposarcoma > are MDM2 negativ, dedifferentiated liposarcoma are positiv. > > > > Most likely the point is "overexpression" of MDM2 protein due to > gene-amplification. Maybe we have to "turn down" our protocol to detect > only > overexpressed MDM2. > > > > Has anyone practical experience with this marker? Can help me? > > > > http://www.proteinatlas.org/ENSG0135679 > > > > > > Bye and thanks in advance > > Gudrun Lang > > ___ > 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] MDM2 on lipsarcoma IHC
Hi all! We try to improve our MDM2-staining and purchased a new antibody. This one stains all nuclei in liposarcoma but also in normal surrounding tissue and lipoma positiv. On the Protein-Atlas-site MDM2 is described with "Ubiquitous nuclear expression". On the other hand I read, that well differentiated liposarcoma are MDM2 negativ, dedifferentiated liposarcoma are positiv. Most likely the point is "overexpression" of MDM2 protein due to gene-amplification. Maybe we have to "turn down" our protocol to detect only overexpressed MDM2. Has anyone practical experience with this marker? Can help me? http://www.proteinatlas.org/ENSG0135679 Bye and thanks in advance Gudrun Lang ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
