This product contains collagenase crude extract and other additives, which can effectively cleave protein hydrogels such as GelMA, SilMA, COMA and collagen, and the enzyme can hydrolyze hydrogel materials without affecting the biological activity of the cells within the hydrogel.

Steps for use:

1. Preparation of 50 mg/mL concentration of laccase standard solution

1) Centrifuge the powder stuck on the inner wall to the bottom of the tube before use.

2) Add 2 mLPBS solution into the tube, shake well to dissolve it, and filter it with 0.22 μm filter tip to remove bacteria.

3) Dispense the prepared lysozyme solution and store it at -20 ℃.

2. Instructions for the use of lytic enzyme solution

1) Prepare 0.3 mg/mL working solution in cell culture medium (i.e., add 6 uL of lysin standard solution to 1 mL of culture medium);

2) Add the appropriate amount of working solution into the plate wells, submerge the hydrogel block, and blow repeatedly with a pipette gun to separate the block from the bottom of the plate and break it sufficiently; the smaller the block is, the faster the speed of cleavage is;

3) Place the well plate in the incubator, 37 ℃ aseptic lysis, every 10-15 min under the microscope to observe the lysis;

4) After the colloid is fully lysed, centrifuge at 1000 rpm for 5 min, discard the supernatant, add 5-10 mL of complete medium and repeat the washing and centrifugation for 1 time.

The obtained cells can be further cultured or analyzed by protein and nucleic acid extraction.

Precautions:

1. the product can be divided after initial dissolution, filtration and sterilization, avoid repeated freezing and thawing.

2. The lysis speed depends on the type of hydrogel, the concentration used and the size of the gel, usually 0.5-2 h can be fully lysed. Usually 0.5-2 h can be sufficiently cleaved. When cleaving larger cell-carrying hydrogel blocks, the blocks should be sufficiently broken, and can be repeatedly blown with a pipette gun or assisted in breaking with sterile forceps, scissors and other instruments.

3. When extracting the cells planted on the surface of the hydrogel, do not break the gel block after adding the working solution, aseptically lysis at 37 ℃ for 5-10 minutes, the cells can be sufficiently separated from the gel block, and the cell suspension can be directly sucked up by centrifugal washing, do not inhale the gel block;

4. Our company is only responsible for the quality control of the material itself, not responsible for the subsequent additional biological experiments and other results unrelated to the quality control of the material itself.

The date of production is shown in the package:

Suzhou Xianjue New Material Technology Co.

Tel: 18068408145

Address: No. 1360, Jinji Lake Avenue, Suzhou, Yue Long Business, 2nd Floor.