Kit used for apoptosis detection of mammal blastocyst cell

Abstract

The invention provides a kit used for apoptosis detection of mammal blastocyst cells. The kit includes: a PBS containing 0.5% of BSA, a PBS containing 2% of paraformaldehyde, a PBS containing 0.5% of Triton X-100 and 0.05% of Tween20, a PBS solution containing 10% of goat serum and 0.05% of Tween20, a rabbit anti-caspase-3 primary antibody, a goat anti-rabbit-IgG secondary antibody having an Alexa Fluor 594 red fluorescein marker, and a PBS containing 20 [mu]M H33342. The kit is mainly based on immunofluorescence technology. A cysteine protease 3 (the caspase-3), which is important in the apoptosis process, is employed as a target point and an antigen-antibody reaction is carried out with the primary antibody aiming to the caspase-3 and the secondary antibody having the fluorescein for carrying out fluorescence labeling and counting to an apoptotic cell in an embryo, and then cell nucleus in all of the cells are stained with a DNA dye so that an apoptosis rate of the blastocyst cells is calculated. With combination of a fluorescence microscope, accurate qualitative and quantitative detection of the apoptotic cell can be carried out. The kit is high in sensitivity, is strong in specificity, is easy to operate, is good in repeatability and is high in efficiency.

Description

technical field [0001] The invention relates to the field of cell biology and immunofluorescence technology, in particular to a kit for detecting apoptosis of mammalian blastocyst cells. Background technique [0002] Apoptosis is a self-death process actively formed by cells in order to maintain a stable internal environment and better adapt to the living environment. It involves the activation, expression and regulation of a series of genes. A large number of light microscopy and ultrastructural research and analysis have proved that apoptosis can be observed in mammalian embryos before implantation. Apoptosis of embryonic cells is conducive to the removal of abnormal cells in embryos. However, the results of the study found that apoptosis Associated with the arrest of embryonic development, apoptosis beyond a certain level is detrimental to embryonic development. Therefore, studying the apoptosis of pre-implantation embryonic cells will help to correctly understand the de...

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Problems solved by technology

However, due to the formation of DNA breakpoints in necrotic cells, TUNEL reaction is also positive; DNA recombination, repair, mitosis in cell metabolism, and human factors in the experimental process, such as sample fixation and protease pretreatment, can cause false positives, so the experiment must be set up. Negative and positive controls

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