2-Color IHC Kit for Mouse/Rat Antibody on Human Tissue, Purple/Red, V2
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)
Rat Primer (RTU)
AP-Polymer anti-Rat (RTU)
Permanent Red Substrate (RTU)
Permanent Red Activator (5x)
Permanent Red Chromogen (100x)
Emerald Chromogen(RTU)
Mounting Medium (RTU)
Rat Primer (RTU)
AP-Polymer anti-Rat (RTU)
Permanent Red Substrate (RTU)
Permanent Red Activator (5x)
Permanent Red Chromogen (100x)
Emerald Chromogen(RTU)
Mounting Medium (RTU)
Usage
The kit supplies the user with two polymer enzyme conjugates: anti-Mouse IgG (minimal cross-reaction to rat) HRP polymer and anti-rat IgG (minimal cross-reaction to mouse) AP polymer. It also includes two distinct substrates/chromogens: Permanent Red and Emerald. The Permanent Red reacts with the anti-Rat AP polymer conjugate to produce the red color, while the Emerald chromogen reacts with the anti-Mouse HRP polymer conjugate to produce the green color. In cases where two proteins are co-expressed in the same location, the area of co-localization appears blue if there is a higher concentration of Emerald and purple-blue if there is a higher concentration of Permanent Red.
Applications
Paraffin tissue (verified), frozen specimen and freshly prepared monolayer cell smears.
Color
BCIP/NBT (purple) and AEC (red)
Specificity
Mouse and Rat
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
To enhance the specificity of antibody staining, a primer step is incorporated into the protocol. The kit employs a non-biotin system, eliminating the need for additional steps to block non-specific binding caused by endogenous biotin.