Kim, We also use the Basalcellcocktail (with p63). The protocol is put together of the two single protocols for the single markers with the usual incubation times. The first HIER step is naturally the retrieval for both antigens. The first part is the dab-kit. After the colour-development a second HIER step takes place. You can choose time and temperature (eg. 60 degrees, 15 min; or 90 degrees 8 min). That depends on how easily your first primary and secondary antibodies can be denatured. Than the Red-kit follows. At the end is counterstain and blueing. If the first primary and sec. ab are'nt denatured you get an overall red result. If the second HIER step is too harsh, you see it in the morphology or high background. Gudrun
-----Ursprüngliche Nachricht----- Von: Artim, Kimberly [mailto:[email protected]] Gesendet: Donnerstag, 22. Jänner 2009 18:02 An: [email protected] Betreff: RE: [Histonet] double staining on the ventana XT Guadrun, Would you be able to provide me more detailed protocol information for the prostate biopsy staining. Are you using P63 as well? Any information you provide would be greatly appreciated. Thank you, Kim Kimberly Artim, HT (ASCP) Technical Coordinator, Anatomic Pathology St Lukes Hospital & Health Network 801 Ostrum Street Bethlehem, PA 18015 610-954-4832 -----Original Message----- From: [email protected] [mailto:[email protected]] On Behalf Of Gudrun Lang Sent: Thursday, January 22, 2009 11:57 AM To: [email protected] Cc: [email protected] Subject: AW: [Histonet] double staining on the ventana XT Hi Jennifer, we perform the doublestain routinly. The preferred protocol is CK HMW/p504s on prostata biopsies. The protocol is easy to establish and the staining results are very good. The run lasts about 4 hours and we do it overnight. You need the Red and the DAB ultraview kit. No extra reagens. Gudrun -----Ursprüngliche Nachricht----- Von: [email protected] [mailto:[email protected]] Im Auftrag von [email protected] Gesendet: Donnerstag, 22. Jänner 2009 01:41 An: [email protected] Betreff: [Histonet] double staining on the ventana XT Greetings, We are looking into doing some immuno double staining. I searched the archives and found a few similar inquiries but no responses. Anyone with experiences to share I would love to hear what you have to say. Everything from what not to do to what works well for you. Many Thanks, Jennifer _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
