Method for detecting glutathione
A glutathione and probe technology, applied in the field of GSH detection, to achieve excellent selectivity and specificity, high selectivity detection, and sensitive fluorescence signal effects
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Embodiment 1
[0020] A method for preparing the GSH fluorescent sensor based on the controllable release technology proposed by the present invention, comprising the steps of:
[0021] (1) Mix 20 μL of washed mercapto magnetic beads with 400 μL of gold nanocage solution evenly, shake at a constant temperature of 37°C for 12 hours, magnetically separate, wash, and add 200 μL of polydiallyldimethyl to the prepared gold nanocage complex Ammonium chloride solution (11.664mg / mL), shake at 37°C for 12h, magnetically separate and wash;
[0022] (2) Add 100 μL rhodamine B solution (1.0×10 -5 mol / L), 10 μL of GSH recognition probe solution (1.0×10 -5 M), shake at 37°C for 12h, magnetically separate, and wash to prepare the GSH fluorescent sensor;
[0023]Wherein, the GSH recognition probe is a nucleotide molecule containing a disulfide bond, and its base sequence is 5'-ACGAGTCA S-S TTT TGT TTG TTC CCC CCT TCT TTC TTA-3'; the sulfhydryl magnetic beads are Commodity purchased (Tianjin Beisi Le Chro...
Embodiment 2
[0025] A kind of method that adopts the GSH fluorescent sensor detection GSH based on controllable release technology that the present invention proposes is as follows:
[0026] (1) Add 10 μL of the GSH sample solution to be tested to the GSH fluorescent sensor based on the controllable release technology proposed by the present invention, dilute it to 100 μL with PBS buffer solution (PH=7.4), shake at a constant temperature of 37°C for 2 hours, and identify the probe and A specific molecular recognition reaction occurs between the target molecules GSH, which leads to the cutting of the disulfide bond, and the recognition probe containing the disulfide bond is detached from the surface of the gold nanocage, thereby opening the molecular gate that blocks the cage hole and dissipating the gold nanocage. The fluorescent molecule rhodamine B in the nanocage is released;
[0027] (2) Magnetic separation, collecting the supernatant and washing liquid for fluorescence detection, dete...
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