Seven Color Multiplex Fluorescent Immunostaining Kit (Anti Rabbit)

The biological significance and clinical value of tissue microenvironment are closely tied to the composition of complex cells within it. The state, abundance, distribution, and phenotype of these cells play a crucial role in understanding the overall dynamics of the microenvironment. It can be shown in situ by antibody staining.The use of immunohistochemical staining is a well-established method in the field of tissue morphology and protein expression analysis within tissues. However, traditional immunohistochemical detection methods have limitations in that they can only detect a single indicator at a time. Consequently, it becomes challenging to comprehensively visualize the cellular composition, state, and interrelationships within the intricate tissue microenvironment. These aspects are of utmost importance for accurate disease diagnosis and effective treatment strategies. Therefore, there is a pressing need to develop advanced techniques that can overcome these limitations and provide a more holistic understanding of tissue biology.

The principle of tyramine amplification technology is quite similar to the DAB color rendering method usually employed for immunohistochemistry. In this technique, a secondary antibody labeled with HRP is used for the detection of the target antigen. The HRP catalyzes the introduction of a fluorescein substrate into the system, which becomes activated and generates a stable covalent bond with the tyrosine present on the antigen. As a result, the final output is a covalently bound fluorescein on the sample, which can be accurately detected using appropriate instrumentation. This technology offers significant advantages over conventional methods and provides unmatched sensitivity and specificity in the detection of various antigens.Following that, the hot repair method was utilized to remove the antibodies that were bound through non-covalent interactions. Subsequently, another round of incubation took place involving the subsequent primary antibody and a different luciferase substrate. This approach allowed for the attainment of multiple labeling, facilitating a more comprehensive analysis.