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Improved method for locating endogenous hormone IAA in bamboo plant tissue

A technology of endogenous hormones and bamboo plants, applied in the field of immunocytochemistry, can solve the problems of background contamination of slices, high requirements for experimental personnel's operation skills, and complicated experimental process, so as to increase the permeability of cell membranes, reduce background interference, The effect of shortening the reaction time

Inactive Publication Date: 2015-07-08
SOUTHWEST FORESTRY UNIVERSITY
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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

Method used

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  • Improved method for locating endogenous hormone IAA in bamboo plant tissue
  • Improved method for locating endogenous hormone IAA in bamboo plant tissue
  • Improved method for locating endogenous hormone IAA in bamboo plant tissue

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Embodiment Construction

[0062] The present invention will be specifically introduced below in conjunction with the accompanying drawings and specific embodiments.

[0063] 1. Experimental samples

[0064] For bamboo shoots of Arrowhead bamboo in Yunnan, the 1st, 3rd, 5th nodes and tops of the shoots that have just been unearthed for about a week (the first segment of the bamboo shoot has not stopped elongating), the first segment of the first segment that has stopped elongating, and a month of unearthed shoots Section 1 of this experiment was used as test samples of different developmental stages in this experiment.

[0065] 2. Experimental Drugs and Formulas

[0066] 1. Pre-fixative solution (2% EDAC, solvent is ultrapure water)

[0067] 2. Post-fixation solution (containing 4% paraformaldehyde and 0.5% glutaraldehyde, concentration is 0.025mol / L, PBS of PH=7.2; Post-fixation solution does not contain sodium cacodylate, avoiding cacodylic acid Sodium hazards to operators while reducing costs)

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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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74G01N1/30
CPCG01N1/30G01N33/74
Inventor 王曙光王雨珺丁雨龙林树燕普晓兰
Owner SOUTHWEST FORESTRY UNIVERSITY
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