2-Color IHC Kit for Goat/Rat Antibody on Human/Mouse Tissue, Geen/Red
In immunohistochemistry, the technique of double staining is widely employed to simultaneously detect and differentiate two distinct antigens within a single tissue sample. The 2-Color IF Kit is designed to use HRP/Emerald (Green) and AP/Permanent Red (red) dye effectively stain 2 different antigens on human and mouse tissue or cell samples when paired with user-supplied goat and rat primary antibodies.
Information
Kit Contents
HRP-Polymer anti-Rat (RTU）
AP-Polymer anti-Goat (RTU）
Permanent Red Substrate (RTU)
Permanent Red Activator (5x)
Permanent Red Chromogen (100x)
Rat Primer (RTU)
Emerald Chromogen (RTU)
Double Stain Blocker
Mounting Medium (RTU)
AP-Polymer anti-Goat (RTU）
Permanent Red Substrate (RTU)
Permanent Red Activator (5x)
Permanent Red Chromogen (100x)
Rat Primer (RTU)
Emerald Chromogen (RTU)
Double Stain Blocker
Mounting Medium (RTU)
Usage
The kit supplies two polymer enzyme conjugates: AP polymer anti-Goat IgG and HRP polymer anti-Rat IgG. These conjugates are paired with two distinct substrates/chromogens: HRP/Emerald (green color) and AP/Permanent Red (red color).
Applications
Paraffin tissue (verified), frozen specimen and freshly prepared monolayer cell smears.
Color
HRP/Emerald (green) and AP/Permanentred (red)
Specificity
Goat and Rat
Tissue Species
Human and Mouse
Storage
2-8℃
Note
The outcome is significantly influenced by several factors, including fixation, tissue slide thickness, antigen retrieval, as well as the dilution and incubation time of the primary antibody. It is crucial for the investigator to carefully evaluate all of these variables and establish the ideal conditions in order to accurately interpret the results.
Principle
The goat and rat primary antibodies will be applied to the tissue simultaneously, but the user will sequentially apply the polymer enzyme conjugate to the specimen. When both proteins are present in the same location, a blue or purple color will develop, depending on the quantity of each screened antigen. If only the anti-goat antigen is present, only the Permanent Red chromogen will be detected. Conversely, if the Rat antigen is present, only the Emerald chromogen will be observed in the cell. The kit is a non-biotin system that prevents non-specific binding of endogenous biotin.