[Histonet] Androgen receptor antibody raised against mouse
Hello all, Does anyone have a recommendation for an anti-mouse Androgen Receptor antibody for Immunohistochemistry on formalin fixed paraffin embedded mouse tissues? We have some mouse prostate tumor models that we're working with. Again we are trying to stain for Androgen Receptor in a mouse, and not the human AR. I have reviewed a few papers and have some decent options, but I'd like to see if the histonet community has any expertise with this. Thank you for all your time and efforts as you always help me out, Steven Swartwood HT (ASCP) Cedars-Sinai Medical Center IMPORTANT WARNING: This message is intended for the use of the person or entity to which it is addressed and may contain information that is privileged and confidential, the disclosure of which is governed by applicable law. If the reader of this message is not the intended recipient, or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this information is strictly prohibited. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Decalcification of mouse jaw and legs
Hello all, I hope everyone is having or had a fantastic week, I'm going to test a few different decal methods on mouse jaws and leg bones. I was wondering if anyone does this routinely out there who is willing to share a protocol. I've never done this on mouse jaw/leg bones before. I've only done this on human tissues. From what I've read it seems that for routine histology low % HCl is fine, but if I wanted to perform IHC then anywhere from 5-15% formic acid may be the best way to go. Nitric acid is probably too strong of a decal agent for these tiny specimens I would assume. Maybe an EDTA based decal agent may be best as well. I'm just really inexperienced with this and I'm very open to ideas and trying a few different methods. Steven Swartwood HT(ASCP) IMPORTANT WARNING: This message is intended for the use of the person or entity to which it is addressed and may contain information that is privileged and confidential, the disclosure of which is governed by applicable law. If the reader of this message is not the intended recipient, or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this information is strictly prohibited. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Rat Neutrophils
Hello again everyone, I have a researcher requesting IHC staining for Rat Neutrophils. I've found a few papers on PubMed and a few antibodies that are specific for Rat neutrophils. The tissue being used is FFPE Rat pancreas and lungs. We are specifically looking for Rat neutrophils. There was no human neutrophils injected into the Rats. I just wanted to see if anyone out there does this or has done this recently and what antibodies they used. A protocol would be greatly appreciated as well. So far I've found more anti-MPO antibodies, but only 1 of them has actual pictures of neutrophil staining in Rats. I was looking into Ly6G antibodies, but none really looked too spectacular. Any and all information is greatly appreciated. I thank all you for your time/efforts for helping me now and the previous help I've received. Steven Swartwood HT(ASCP) Cedars Sinai Medical Center steven.swartw...@cshs.org IMPORTANT WARNING: This message is intended for the use of the person or entity to which it is addressed and may contain information that is privileged and confidential, the disclosure of which is governed by applicable law. If the reader of this message is not the intended recipient, or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this information is strictly prohibited. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] FoxP3 and Geminin Immunohistochemistry
Hello all, I've done some research on Foxp3 and Geminin and I've seen a few antibodies that look sufficient for IHC in FFPE tissues. My lab is currently looking into purchasing these antibodies for a few different projects. Does anyone on here use these two antibodies for IHC in FFPE tissues? I'm going to use them in bright field, but we may also turn to IF for other studies. I'd be grateful for any information from what clone/company do you buy from and retrieval methods to primary antibody incubations/dilutions. Thank you all for any information, Steven Swartwood HT(ASCP) Cedars Sinai Medical Center steven.swartw...@cshs.org IMPORTANT WARNING: This message is intended for the use of the person or entity to which it is addressed and may contain information that is privileged and confidential, the disclosure of which is governed by applicable law. If the reader of this message is not the intended recipient, or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this information is strictly prohibited. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] ZDHHC11
Hello Histonetters, This is all pertaining to staining human tissues, I work in a research core lab and was wondering if anyone had IHC-P (FFPE) experience in staining for ZDHHC11? From the data our computational team has pulled 70% of primary prostate ca is positive and 90% of metastatic prostate ca is positive. There's not much information out there for IHC-P staining with this antibody. The researcher that wants this staining done bought this from Thermo Scientific and they don't have much information either... other than a picture of lung adenocarcinoma that about 4 other companies also show from the same antibody. One company advised using pepsin digestion along with citrate retrieval and another said tris-EDTA. I figure to try out multiple retrievals (as usual with new antibodies), but has anyone ever used this antibody? I was trying to find some good control tissue as well (besides the cancer tissues) and all I can find is normal ovary so far. We have a Discovery Ultra here and I've tried a titer with a few different high grade primary prostate cancer tissues with different HIER and was unsuccessful, but I've yet to try the enzymatic approach. Any help would be tremendously appreciated, Steven Swartwood HT(ASCP) Cedars Sinai Medical Center steven.swartw...@cshs.org IMPORTANT WARNING: This message is intended for the use of the person or entity to which it is addressed and may contain information that is privileged and confidential, the disclosure of which is governed by applicable law. If the reader of this message is not the intended recipient, or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this information is STRICTLY PROHIBITED. If you have received this message in error, please notify us immediately by calling (310) 423-6428 and destroy the related message. Thank You for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Biocare Decloaker Nx Gen
Has anyone ever used the Biocare Decloaking Chamber Nx Gen for epitope unmasking (heat retrieval)? Also does anyone have any personal pros/cons with this instrument? We are thinking about getting one because of inconsistencies with our current method using a vegetable steamer. We use many different tissue types both animal and human. Also we use many different antibodies. Currently with the vegetable steamer we've used Citrate, EDTA, and Tris-EDTA combinations. We've used times from 30-60 minutes, and even tried 30 minutes x 2 or 40 minutes x 2, but we still see some inconsistent staining. To make sure it wasn't due to fixation vimentin staining was performed and shows even staining. It's just certain antibodies that are giving us trouble even from multiple different companies. Any advice would be greatly appreciated and very beneficial to our work pertaining to the instrument. Thanks again everyone have a great end of the week, Steven Swartwood HT(ASCP) steven.swartw...@cshs.org IMPORTANT WARNING: This message is intended for the use of the person or entity to which it is addressed and may contain information that is privileged and confidential, the disclosure of which is governed by applicable law. If the reader of this message is not the intended recipient, or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this information is STRICTLY PROHIBITED. If you have received this message in error, please notify us immediately by calling (310) 423-6428 and destroy the related message. Thank You for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Double staining with IF/IHC - chromogenic
Hello everyone, I have a co-worker in the research department that would like to know if anyone has ever combined and IF stain with chromogenic IHC staining. His theory for this is that IF shows better nuclear staining from an image analysis stand point. If you have heard of this can you send me literature on it, or if you've done this what is your protocol? It would seem that doing IF first than IHC might cause quenching of the IF, but then again doing chromogenic staining first might cover the IF antigen. I suspect that only an chromogenic cytoplasmic IHC stain can be combined with a nuclear IF marker with IHC first. Any ideas? Again this is specifically from an image analysis stand point. Steven Swartwood HT(ASCP) Cedars Sinai Medical Center steven.swartw...@cshs.org IMPORTANT WARNING: This message is intended for the use of the person or entity to which it is addressed and may contain information that is privileged and confidential, the disclosure of which is governed by applicable law. If the reader of this message is not the intended recipient, or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this information is STRICTLY PROHIBITED. If you have received this message in error, please notify us immediately by calling (310) 423-6428 and destroy the related message. Thank You for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
