I-kiti ye-2-color multiplex IHC Plus (TYR520/I-antibody yesibili yesikhala se-rabbits no-mice)|Neorise Biotech Co.,Ltd

Imininingwane enzuzo

I-MOQ:1

Indlela yokuthumela:I-express, I-ihlathi

inombela yokucaciswa:100T

Ukubonisa umlingani

Principle Introduction:

The principle of Multiplex Immunohistochemistry (mIHC) is that the antigen on tissue sections specifically binds to the primary antibody. Then, a secondary antibody conjugated with horseradish peroxidase (HRP) is added to bind with the primary antibody. After adding the tyramine fluorescence substrate, non-activated tyramine fluorescein substrates are activated by HRP-catalyzed hydrogen peroxide. Upon addition of a fluorescein substrate, the non-activated tyramine fluorescein substrate is catalytically activated by HRP in the presence of hydrogen peroxide. The activated substrate covalently binds to nearby protein residues, including tryptophan, histidine, and tyrosine, allowing for stable antigen-fluorescein binding.

In multiplex labeling experiments, the non-covalently bound primary and secondary antibodies from the previous round can be washed away using heat repair or an antibody eluent. Subsequently, the next round of primary-HRP secondary antibody is incubated, followed by the application of a different tyrosine fluorescein substrate. This process can be repeated to achieve multiple labeling steps. Since only one antibody is incubated at a time in each system, there is no concern about antibody cross-reactivity or the need to match primary and secondary antibodies, significantly reducing the challenges associated with selecting and matching antibodies from different species in experimental design.

The kit contains specific tyramine fluorescent dyes, which include one or more of the following: TYR-480Plus, TYR-520Plus, TYR-570Plus, TYR-620Plus, TYR-690Plus, and TYR-780Plus. These fluorescent dyes can be used individually or in combination, allowing for single, double, triple, or even more complex fluorescence amplification and homologous antibody labeling, greatly enhancing the versatility the kit.

Our Multi Fluorescence Staining Plus(mIHC) Kits can currently achieve up to 9 colors. The dye combinations are detailed in the table below:

Kit components:

TYR-***Plus Fluorescent Dyes, TSA buffer, HRP goat anti mouse/rabbit universal secondary antibody, DAPI Staining Solution (Ready-to-use), Antibody Diluent, 3% hydrogen peroxide.

Advantages of Multi Fluorescence Staining Plus(mIHC) Kits:

1. High sensitivity, 10-50 times higher than conventional TSA assay kits,

2. Be careful not to have too strong a signal; If the signal is too strong, the dye concentration/reaction time/primary antibody concentration will be reduced,

3. If the signal is too low, increase the dye concentration/reaction time/primary antibody concentration;

4. The higher the concentration of the Multi Fluorescence Staining Plus(mIHC) Kits, the exponentially enhanced the signal; 1: 50 can obtain the strongest signal, and 1:500 is equivalent to the signal of a conventional TSA kit.

5. The Multi Fluorescence Staining Plus(mIHC) Kit has strong quenching resistance to fluorescent dyes, and does not require exposure to sunlight throughout the entire process. It also does not require operation in a dark environment during use, but cannot be exposed to sunlight.

Operational staining procedure:

Example of imaging using our Multi Fluorescence Staining Plus Kits:

(A) Human fallopian tube tissue index A (570 yellow)+index B (520 green)+index C (690 powder);

(B) Mouse kidney tissue index A (570 yellow)+index B (520 green)+index C (690 powder);

(C) Human gastric cancer tumor indicator A (690 powder)+indicator B (620 red)+indicator C (570 yellow)+indicator D (520 green); (D) Human intestinal tissue index A (570 yellow)+index B (520 green)+index C (690 pink)+index D (620 red)+index E (480 green).

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