2-Color IHC Kit for Mouse/Rabbit Antibody on Human 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 Emerald (green) and Permanent Red (red) dye effectively stain 2 different antigens on human tissue or cell samples when paired with user-supplied mouse and rabbit primary antibodies.
Information
Kit Contents
HRP-Polymer anti-Mouse (RTU)
AP-Polymer anti-Rabbit (RTU)
Permanent Red Substrate (RTU)
Permanent Red Activator (5x)
Permanent Red Chromogen (100x)
Emerald Chromogen (RTU)
Mounting Medium (RTU)
AP-Polymer anti-Rabbit (RTU)
Permanent Red Substrate (RTU)
Permanent Red Activator (5x)
Permanent Red Chromogen (100x)
Emerald Chromogen (RTU)
Mounting Medium (RTU)
Usage
The kit supplies two polymer enzyme conjugates: HRP-Polymer anti-Mouse IgG and AP-Polymer anti-Rabbit IgG. It also includes two chromogens: Emerald (green) and Permanent Red (red).
Applications
Paraffin tissue (verified), frozen specimen and freshly prepared monolayer cell smears.
Color
Emerald (green) and Permanentred (red)
Specificity
Mouse and Rabbit
Tissue Species
Human
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
This protocol offers a convenient approach by simultaneously applying the enzyme conjugates to the specimen. The kit enables researchers to visualize colocalization of two proteins through a color change when the chromogens overlap, allowing for semi-quantitative analysis. For example, if the co-localization area stains blue, it indicates a higher concentration of the antigen indicated by Emerald in the cell, while a purple color indicates a higher concentration of the antigen indicated by Permanent-Red.
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