The TripleStain IHC Kit (ab183298) is designed to be used with user-supplied mouse, rat and rabbit primary antibodies to detect three distinct antigens on mouse tissue and cell samples or human tissue. This kit has been tested on paraffin embedded tissue, however it can also be used on frozen tissue or cell smears.
Triple staining is a common method used in immunohistochemistry for the detection of three distinct antigens and their co-expression in a single tissue. Abcam’s TripleStain IHC Kit (ab183198) supplies polymer enzyme conjugates: mouse HRP polymer, Rabbit HRP polymer, and Rat AP-polymer with three substrates/chromogens; DAB (brown), DAB-Ni (black), and Permanent Red (Red). The TripleStain Kit is a non-biotin system avoiding non-specific background due to endogenous biotin binding. This kit has been optimized to have no cross detection when detecting two primary antibodies from the mouse and rat host species using a unique blocking system. Simplified steps allow the user to complete triple staining within 5 hours (without antigen retrieval) or 6 hours (with antigen retrieval). This protocol also includes a method to dehydrate, clear and permanently mount slides with coverslip.
|Components||24 ml||72 ml||240 ml|
|Antibody Blocker (40x)||2 x 15ml||1 x 50ml||1 x 100ml|
|Aqueous Mounting Medium||1 x 15ml||2 x 18ml||1 x 120ml|
|DAB Chromogen (20x)||1 x 2ml||1 x 4ml||1 x 12ml|
|DAB Substrate||1 x 15ml||2 x 18ml||1 x 120ml|
|DAB-Ni Substrate (20x)||1 x 1ml||1 x 2ml||1 x 6ml|
|Hydrogen Peroxide (20x)||1 x 1ml||1 x 2ml||1 x 6ml|
|Mouse HRP Polymer||1 x 12ml||2 x 18ml||1 x 120ml|
|Nickel Solution (7x)||1 x 3ml||1 x 6ml||1 x 18ml|
|Permanent Red Activator (5x)||1 x 3ml||1 x 7.2ml||2 x 12ml|
|Permanent Red Chromogen (100x)||1 x 150µl||1 x 360µl||1 x 1.2ml|
|Permanent Red Substrate||1 x 15ml||2 x 18ml||1 x 120ml|
|Rabbit HRP Polymer||1 x 6ml||1 x 18ml||1 x 60ml|
|Rat AP Polymer||1 x 6ml||1 x 18ml||1 x 60ml|
|Rat Primer||1 x 12ml||2 x 18ml||1 x 120ml|
Our Abpromise guarantee covers the use of ab183298 in the following tested applications.
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
ab183298 has not yet been referenced specifically in any publications.