2-Color IHC Kit for Mouse/Rabbit Antibody on Rodent Tissue, Purple/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 BCIP/NBT (Purple) and AEC (Red) dye effectively stain 2 different antigens on mouse tissue or cell samples when paired with user-supplied mouse and rabbit primary antibodies.
Information
Kit Contents
Mouse Primer (RTU)
HRP-Polymer anti-Rabbit (RTU)
AP-Polymer anti-Mouse (RTU)
BCIP/NBT Chromogen(RTU)
AEC Substrate (20x)
AEC Chromogen (20x)
Hydrogen Peroxide (20x)
Mounting Medium (RTU)
HRP-Polymer anti-Rabbit (RTU)
AP-Polymer anti-Mouse (RTU)
BCIP/NBT Chromogen(RTU)
AEC Substrate (20x)
AEC Chromogen (20x)
Hydrogen Peroxide (20x)
Mounting Medium (RTU)
Usage
The kit provides two polymer enzyme conjugates: Mouse AP Polymer and Rabbit HRP Polymer, each with its own specific substrates/chromogens. The Mouse AP Polymer uses BCIP/NBT, which produces a purple color, while the Rabbit HRP Polymer uses AEC, resulting in a red color.
Applications
Paraffin tissue (verified), frozen specimen and freshly prepared monolayer cell smears.
Color
BCIP/NBT (purple) and AEC (red)
Specificity
Mouse and Rabbit
Tissue Species
Rodent
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
To enhance the specificity of antibody staining, a primer step is employed. In this step, both enzyme conjugates are simultaneously applied to the specimen and mixed on the slide. The kit utilizes a non-biotin system to prevent non-specific binding of endogenous biotin.