Abcam's TripleStain IHC Kit (ab183295) is designed to be uses with user-supplied one mouse and two rabbit primary antibodies to detect three distinct antigens on a single mouse/rat tissue or cell sample. This kit has been tested on tissue specimens that are paraffin embedded; however it may also be used on frozen or freshly prepared monolayer cell smears.
Triple staining uses traditional methods in immunohistostaining to reveal three distinct antigens and their co-expression on a single tissue. Abcam's TripleStain IHC Kit (ab183295) supplies polymer enzyme conjugates: Mouse HRP Polymer, Rabbit HRP Polymer and Rabbit AP Polymer with three substrates/chromogens; DAB (brown), Emerald (green), and Permanent Red (Red). Abcam's TripleStain IHC Kit (ab183295) is a non-biotin system, avoiding non-specific binding caused by endogenous biotin. This kit has been optimized to have no cross detection when detecting two primary antibodies from the same host species using a unique blocking system. Simplified steps allow the user to complete triple staining within 5 hours (without antigen retrieval) or 6-7 hours (with antigen retrieval). The well tested protocol allows the user to permanently mount slides with coverslip.
|Components||Identifier||240 ml||72 ml||24 ml|
|Non-aqueous Mounting Medium||—|
|Antibody Blocker (40x)||3 x 100ml||1 x 100ml||2 x 15ml|
|Blocker A||1 x 120ml||2 x 18ml||1 x 12ml|
|Blocker B||1 x 120ml||2 x 18ml||1 x 12ml|
|DAB Chromogen (20x)||1 x 6ml||1 x 2ml||1 x 1.5ml|
|DAB Substrate||1 x 120ml||2 x 18ml||1 x 15ml|
|Emerald Chromogen||1 x 120ml||2 x 18ml||1 x 12ml|
|Mouse HRP Polymer||1 x 60ml||1 x 18ml||1 x 6ml|
|Mouse Primer||1 x 120ml||2 x 18ml||1 x 12ml||2 x 18ml||1 x 12ml|
|Permanent Red Activator (5x)||1 x 12ml||1 x 7.2ml||1 x 3ml|
|Permanent Red Chromogen (100x)||1 x 1.2ml||1 x 360µl||1 x 150µl|
|Permanent Red Substrate||1 x 120ml||2 x 18ml||1 x 15ml|
|Rabbit AP Polymer||1 x 60ml||1 x 18ml||1 x 6ml|
|Rabbit HRP Polymer||1 x 120ml||2 x 18ml||1 x 12ml|
Our Abpromise guarantee covers the use of ab183295 in the following tested applications.
|IHC-P||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
ab183295 has not yet been referenced specifically in any publications.