Method for detecting galectin-1 in blood and reagent used
A technology for galectin and blood, which is applied in the field of detecting galectin-1 in blood, can solve the problems of low absolute content, lack of detection methods, and little help in early detection of cancer metastasis and intervention treatment, and achieves high sensitivity Effect
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Embodiment 1
[0052] Example 1, Preparation of lactose-modified gold nanorod detection probes
[0053] In this example, the seed-induced gold nanorod synthesis method is used to prepare gold nanorods with good dispersion performance, single particle size, suitable long-short axis ratio, and stable existence; synthesize suitable lactose ligands; Lactose ligands were loaded onto the surface of gold nanorods to achieve lactosylation on the surface of gold nanorods, and a lactose-modified gold nanorod detection probe was prepared and its structure was confirmed.
[0054] 1. Preparation of gold nanorods
[0055] Gold rods were prepared by seed-induced synthesis of gold nanorods in aqueous solution. First reduce the chloroauric acid with sodium borohydride to prepare 1-2nm gold seeds, then add the gold nano-seeds to the chloroauric acid (HAuCl 4 ), silver nitrate (AgNO 3 ), dilute hydrochloric acid (HCl), ascorbic acid, and dodecyldimethylammonium bromide (CTAB) in a mixed aqueous solution to ...
Embodiment 2
[0080] Example 2, the detection of galectin-1 by molecular level gold nanorods (Lac-GNRs)
[0081] This example evaluates the detection of galectin-1 at the molecular level.
[0082] The method used to detect galectin-1: In operation, add 100 microliters of the substance to be tested (aqueous solution of galectin-1) to 1 milliliter of the Lac-GNR aqueous solution prepared in Example 1, shake well and let stand at room temperature for one Hours, use a UV-Vis-NIS spectrophotometer (UV-Vis-NIS spectrophotometer) to optically detect the system, measure the light absorption from 250nm to 1100nm, and judge whether the test substance contains galectin-1 through the change of the absorption curve, It can even be judged by the naked eye through the change in color.
[0083] The detection result of this embodiment finds that Lac-GNRs can detect the existence of galectin-1 extremely sensitively, and the detection limit can reach 1pM, and the response value changes with the change of gal...
Embodiment 3
[0085] Example 3. Detection of galectin-1 by Lac-GNRs at the cellular level
[0086] In this example, the detection of galectin-1 is studied at the cellular level, because galectin-1 is a secreted protein, which will be secreted into the extratumor matrix to play a role. The cancer cells are cultured by the method of cell culture, and the culture fluid thereof is taken for detection, and the existence of galectin-1 should be detected.
[0087] In this example, HUH-7 (human liver cancer cells), MCF-7 (human breast cancer cells), JEG-3 (human choriocarcinoma cells), HeLa (human cervical cancer cells), Hep-G2 (human liver cancer cells) were cultured in total. Cells), C6 (mouse glioma cells) and other six kinds of tumor cells, and their cell culture fluids were detected. The culture conditions of the cells are as follows: the cells are planted in H-DMEM culture medium containing 10% fetal bovine serum (high-glucose Dulbecco's minimum essential medium, the cell concentration is ab...
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