Improved method for locating endogenous hormone IAA in bamboo plant tissue

Abstract

The invention discloses an improved method for locating endogenous hormone IAA in bamboo plant tissue, ans aims to improve the existing immunogold-silver staining method. TBS is used as a buffer solution and solvent in the experiments to substitute PBS, TBS does not generate precipitate with calcium ions, and has extremely low interference on biological reaction, thus reduce some nonspecific coloration background interference and improve the accuracy of the results; by adding gelatin in the color liquid, chromogenic time is basically controlled in 15-20 min, and time is enough to check the chromogenic level in the chromogenic period, so as to obtain good repetition rate; and incubation of secondary antibody at room temperature shortens the reaction time. The method of the invention not only streamlines reaction steps, and has mild reaction conditions and easy control of coloration, can obtain more elegant and accurate image, and has good application and popularization value in the immune localization of bamboo plant endogenous hormone.

Description

technical field [0001] The invention relates to a method for specifically locating endogenous hormone IAA in bamboo plant tissue, in particular to a method for locating endogenous hormone IAA in plant tissue by using an improved immunogold and silver staining technique, and belongs to the field of immunocytochemistry. Background technique [0002] The immunogold and silver staining method is used to localize the endogenous hormone IAA in plant tissue. It is based on paraffin section, using antigen-antibody specific reaction, and the secondary antibody with colloidal gold particles is specific to the plant tissue section that has been incubated with the primary antibody. reaction, so that the colloidal gold particles are marked in the tissue where there are plant endogenous hormones, and finally the color is developed with a color solution containing silver nitrate. The principle of color development is that silver nitrate is reduced to silver atoms under the action of reduci...

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Problems solved by technology

[0029] Although this paper also carried out immunolocalization on the basis of paraffin sections, the experimental process was very complicated and the operation was cumbersome.

Although the experiment used a 37°C reaction environment to shorten the reaction time during antigen recovery, primary and secondary antibody incubation, and urea digestion, but due to many operational steps and many drugs involved, it is easy to cause background contamination of the slices, and it is harmful to the experimenters. High operational skill requirements

[0030] Moreover, during the material fixation process, the experiment used sodium cacodylate buffer as a lotion, and sodium cacodylate is a precursor drug, which is harmful to operators and expensive

[0031] In addition, the developing solution in this paper contains 2% gelatin, but in the development process of the endogenous hormone gold and silver method of bamboo shoots, the developing solution containing 2% gelatin cannot develop color normally, and the background non-specific color development is serious

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