2-Color IHC Kit for Mouse/Rabbit Antibody on Human 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 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)
BCIP/NBT (RTU)
AEC Substrate (20x)
AEC Chromogen (20x)
Hydrogen Peroxide (20x)
Mounting Medium (RTU)
AP-Polymer anti-Rabbit (RTU)
BCIP/NBT (RTU)
AEC Substrate (20x)
AEC Chromogen (20x)
Hydrogen Peroxide (20x)
Mounting Medium (RTU)
Usage
The kit provides the user with two polymer enzyme conjugates: an HRP Polymer anti-Mouse IgG and an AP-Polymer anti-Rabbit IgG. Each enzyme is accompanied by reactive chromogens. The HRP Polymer anti-Mouse IgG is used with the AEC chromogen (Red Brick color), while the AP-Polymer anti-Rabbit IgG is used with the BCIP/NBT chromogen (Purple/Blue 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
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
By applying the enzyme conjugates and chromogens as a mixture, simplified steps in this protocol significantly speed up the process. The enzyme conjugated polymers and chromogens are carefully optimized to produce a strong signal without any background interference. Additionally, the kit utilizes a non-biotin system, eliminating the need to block endogenous biotin and preventing non-specific binding.
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