Paraffin section immunofluorescence multiple staining kit and use method thereof

Abstract

The invention provides a paraffin section immunofluorescence multiple staining kit. The kit comprises a reagent A, a reagent B, a diluent, an FITC reagent, a Cy3 reagent and a Cy5 reagent. The invention further provides a use method of the paraffin section immunofluorescence multiple staining kit. According to the kit, a TSA signal amplification technology is adopted, and the principle is similarto that of common immunohistochemistry; after a primary antibody is combined with an antigen, an HRP labeled secondary antibody is incubated; by catalyzing a fluorescein substrate added into the system by HRP, an activated fluorescent substrate is generated, wherein the activated substrate can be covalently combined with tyrosine on the antigen, so that the fluorescein is stably covalently bondedon the sample; then, a non-covalently bound antibody is washed away by thermal remediation, a second primary antibody is used for second incubation, another fluorescein substrate is used for replacement, and multiple labeling can be realized by repeating the steps; only one antibody is incubated in each round; and the non-covalently bound antibody is washed away after microwave treatment, so thatthe species source of the used antibody is not limited.

Description

technical field [0001] The invention relates to the field of biomedical application technology, in particular to a paraffin section immunofluorescence multiple staining kit and a use method. Background technique [0002] Immunofluorescence technology is an experimental technique based on the specific combination of antibodies and antigens. It is widely used in basic research and clinical experiments due to its advantages of simple operation, high efficiency, and strong specificity. With the rise of immunotherapy, more and more More and more markers in the tumor microenvironment have been found to be closely related to disease treatment and progression. Simultaneously staining and marking these markers and studying their co-localization, expression and distance relationship have become a common demand for tumor immunity research ;Traditional immunofluorescent labeling can achieve simultaneous labeling of two antibodies, and the two antibodies need to be prepared from differen...

AI Technical Summary

Problems solved by technology

[0002] Immunofluorescence technology is an experimental technique based on the specific combination of antibodies and antigens. It is widely used in basic research and clinical experiments due to its advantages of simple operation, high efficiency, and strong specificity. With the rise of immunotherapy, more and more More and more markers in the tumor microenvironment have been found to be closely related to disease treatment and progression. Simultaneously staining and marking these markers and studying their co-localization, expression and distance relationship have become a common demand for tumor immunity research ;Traditional immunofluorescence labeling can achieve simultaneous labeling of two antibodies, and the two antibodies need to be prepared from different species to be connected to secondary antibodies of different fluorescein for color development; this experimental method requires the source species of the primary antibody High, if you want to label two or more antigens, all antibodies are required to come from different species, and often you cannot purchase all the antibodies that meet the requirements, resulting in the inability to perform multiple antigen labels

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