2-Color IHC Kit for 2 Mouse 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 2 mouse primary antibodies.
Information
Kit Contents
HRP-Polymer anti-Mouse (RTU)
AP Polymer anti-Mouse (RTU)
Permanent Red Substrate (RTU)
Permanent Red Activator (5x)
Permanent Red Chromogen (100x)
Antibody Blocker (40x)
Double Mouse Blocker 1 (RTU)
Double Mouse Blocker 2 (RTU)
Emerald Chromogen (RTU)
Mounting Medium (RTU)
AP Polymer anti-Mouse (RTU)
Permanent Red Substrate (RTU)
Permanent Red Activator (5x)
Permanent Red Chromogen (100x)
Antibody Blocker (40x)
Double Mouse Blocker 1 (RTU)
Double Mouse Blocker 2 (RTU)
Emerald Chromogen (RTU)
Mounting Medium (RTU)
Usage
The kit supplies two polymer enzyme conjugates: HRP polymer anti-Mouse IgG and AP polymer anti-Mouse IgG. It offers two different substrates/chromogens: Emerald (Green color, to be used with HRP polymer anti-Mouse IgG) and Permanent-Red (Red color, to be used with AP polymer anti-Mouse IgG).
Applications
Paraffin tissue (verified), frozen specimen and freshly prepared monolayer cell smears.
Color
Emerald (green) and Permanentred (red)
Specificity
Mouse
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
The use of simplified steps in this protocol results in a faster procedure, while the blocking buffers effectively prevent false positives when using two primary antibodies from the same host species. By utilizing this kit, researchers are able to visually determine the co-localization of two proteins based on the color change resulting from the overlapping of chromogens. This color change can provide a semi-quantitative measure. For instance, if the co-localized area stains blue, it indicates that the antigen indicated by Emerald is present in higher concentrations within the cell. Conversely, if the color appears purple, it suggests that the antigen indicated by Permanent-Red is expressed at higher concentrations. Additionally, the kit utilizes a non-biotin system to avoid nonspecific binding of endogenous biotin.