Five Color Multiplex Fluorescent Immunostaining Kit (Mouse Rabbit)

The tissue microenvironment contains intricate cell components, each with their own phenotype, state, abundance, and distribution. Understanding these cells is crucial for their biological and clinical significance. Immunohistochemical staining with antibodies provides a way to visualize and study tissue morphology and protein expression in situ. However, conventional immunohistochemistry is limited to displaying a single index and cannot fully capture the composition, state, and relationships within the complex tissue microenvironment, which is vital for disease diagnosis and treatment.
To address this limitation, tyrosine signal amplification (TSA) technology has been developed. Similar to the conventional DAB coloring method, TSA utilizes HRP-labeled secondary antibodies. HRP catalyzes the addition of fluorescent substrates, which, when activated, covalently bind to the tyrosine residues on the antigens within the sample. This covalent binding ensures that the sample is permanently labeled with fluorescence. The non-covalently bound antibodies are then washed away using heat repair methods, allowing for another round of incubation with a different primary antibody and a new set of fluorescent substrates. This process can be repeated, enabling multiple labels and facilitating a more comprehensive understanding of the cell composition and relationships within the tissue microenvironment.
By utilizing tyrosine signal amplification technology, researchers can overcome the limitations of traditional immunohistochemistry and gain valuable insights into the complex cellular dynamics and interactions in tissues. This information is crucial for advancing disease diagnosis and treatment strategies.

The primary application of this product is for the immunohistochemical labeling of human or mouse tissues, paraffin sections, and TMA chips. To create similar content, we could say that this item is commonly employed in the staining of samples from both human and mouse subjects, which may include tissues, paraffin sections, and TMA chips.

To prepare a fluorescent dye solution, there are different methods depending on whether the dye is in dry powder or liquid form. If the dye is in dry powder form, dissolve it in 100ul of DMSO to create a concentrated 100X dye mother liquor. However, if the dye is already in liquid form, it means it was dissolved in DMSO during the manufacturing process to create a 100X mother liquor. In this case, the dye can be directly used.
To create a dye working solution for immediate use, dilute the dye with the signal amplification reaction solution at a 1:100 ratio. This will ensure the dye is appropriately diluted and ready for use.
For DAPI, the process is slightly different. Dilute the DAPI dye with sterile water at a ratio of 1:100 to prepare a working solution. This solution can then be used for the intended purpose.
To ensure accurate results in your experiments, it is important to carefully consider the use of the secondary antibody. The pika universal secondary antibody provided in the kit is specifically designed for human samples, but may not be suitable for mouse tissue. It is crucial to verify that the species of your primary antibody matches before proceeding with the experiment. If working with mouse tissues, it is necessary to use other types of secondary antibodies. This adjustment will help to ensure that your research is based on accurate and reliable data.

1. The primary use of this kit is for immunohistochemistry and it is not suitable for any other applications.
2. Only professionals with appropriate training and expertise should handle this kit.
3. It is crucial to adopt proper protective measures to prevent any contact of the reagents with the skin and eyes.
4. The effectiveness of the reagents may decrease if used beyond their expiration date, so it is important to avoid using expired reagents.
5. Mixing the dyeing components of this kit with products from other companies may lead to abnormal results during the dyeing process.
6. Insufficient dewaxing can have a negative impact on the overall dyeing outcome.
7. To ensure accurate results, it is essential to simultaneously include a positive control and a negative control during the experiment, minimizing the chances of false negative and false positive outcomes.
8. All waste generated during the usage of this kit must be handled in strict accordance with the regulations outlined in the "Medical Waste Management Regulations."

Fluorescent dyes should be stored in the dark at 2～8° C. The validity period is 12 months from receipt.

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4. Qian Bangguo. Jiao Lei. The application of multi-label immunofluorescence staining and Doppler imaging technology in histological research. Chinese Journal of Histochemistry and Cytochemistry, 2017 (4): 373-382.