A kind of mixture for improving memory and its preparation method and application
A mixture, memory technology, applied in the field of molecular biology, to achieve the effect of improving cognition, enhancing memory, and reducing drug dependence
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[0067] In the second aspect, the present invention provides a method for preparing the mixture HC4201612, the mixture is prepared from plasma extraction, and the preparation method sequentially includes the following steps: plasma collection, low temperature filtration, low temperature ultrafiltration, cold ethanol precipitation, SD inactivation , centrifugation, anion exchange, dialysis, concentration steps. details as follows:
[0068] Step 1. Plasma collection:
[0069] The blood is collected with an anticoagulant tube, and the supernatant is collected by centrifugation to obtain plasma; preferably, the centrifugation speed is 500-1500g, the centrifugation time is 10-30min, and the centrifugation temperature is 0-4°C.
[0070] Exemplarily, the above-mentioned centrifugal speed can be any value among 500g, 1000g, 1250g, 1500g, etc. or any value range between any two, and the above-mentioned centrifugation time can be any value among 10min, 15min, 20min, 30min, etc. or any t...
Embodiment 1
[0124] The present embodiment is the preparation method of mixture HC4201612, and this method comprises the following steps:
[0125] (1) Collection of plasma
[0126] The blood of healthy people is obtained from blood donation in the hospital (the blood collection tube is an anticoagulant tube). During the blood collection process, the blood collection tube is shaken to prevent blood coagulation. Put the blood collection tube into the centrifuge, set the rotation speed to 1500g, centrifuge for 15 minutes, and the centrifugation temperature is 4°C. Then use a pipette to carefully aspirate the supernatant, which is the collected human plasma.
[0127] (2) Low temperature filtration:
[0128] The collected plasma was added to a filter with a filter membrane pore size of 0.22 microns, and a peristaltic pump was used to apply a pressure of 10 MPa for filtration. During the process, the temperature of the entire filtration device was controlled at 0-4° C. to obtain a filtrate.
...
Embodiment 2
[0144] In this example, the mixture HC4201612 prepared in Example 1 was identified by SDS-PAGE denaturing gel electrophoresis. The identification method includes the following steps:
[0145] (1) Take 2 microliters from the above-mentioned mixture HC4201612, and measure its absorbance under ultraviolet 280 nm, thereby calculating the protein concentration of the mixture HC4201612.
[0146] (2) Take a certain volume of the mixture HC4201612 and mix it with 1 microliter of 5× protein loading buffer (purchased from Beijing Lamboride Biotechnology Co., Ltd., Cat. No. D621), which is the sample to be electrophoresed. The sample contains 10 micrograms of protein.
[0147] (3) Heat the electrophoresis sample to 100°C, heat for 20 minutes to denature the protein, then immediately put the sample on ice, wait for 5 minutes and start running SDS-PAGE denatured reducing gel (the preparation method of the SDS denatured reducing gel is as follows : 30 (w / v)% acrylamide Acr-Bis (purchased ...
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