1. Centrifuge: Centrifugal (10, 000 rpm) for 20-30 seconds or low speed centrifugation (2,000 rpm) for 5 minutes.
2. Reconstitution: Dissolve in deionized water or other buffer (selected according to the instructions) to the concentration required by the instructions (typically 0.1-1.0 mg/ml). Do not oscillate when dissolved, to avoid protein inactivation due to chemical bond cleavage, gently shake with a suitable solvent and let stand for a period of time, after the cytokine is completely dissolved. Solvent selection:
1) For products that require deionized water to be dissolved, the salt solution must not be used to avoid irreversible precipitation of proteins due to excessive salt concentration;
2) For products that require dissolution with a salt solution, please follow the concentration on the instruction sheet to avoid excessive salt concentration.
3. Aliquot and Storage: After protein solubilization, it can be further diluted into working fluid according to your own experiment. Dilute with RPMI 1640, DMEM or PBS. The working solution can be stored at 4 ° C for 1 week. For long-term storage, it is best to add 10% FCS (calf or fetal bovine serum) or 0.1% BSA (bovine serum albumin) or 5% HAS to the dilution. Human serum albumin) is used to stabilize the protein, which is then stored frozen at -20 ° C (Note: not frozen below -50 ° C). The volume of the working fluid in each tube at the time of dispensing is preferably the amount of one experiment, so that one working fluid is used in each experiment to avoid the decrease in protein activity caused by repeated freezing and thawing.
The amount of cytokines is extremely small, so rigorous operation is necessary, and any improper dissolution will cause the experiment to fail, resulting in a lot of waste. Please be cautious!
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