本产品与组织消化酶 (货号O1521148) 配套使用,包含多种营养因子,能够有效避免组织在消化过程中过 度损伤,提高组织消化得到的原代细胞活力。消化得到的细胞悬液可用于原代细胞培养以及利用流式细胞仪进行多种 细胞生物学实验,如凋亡、增殖、细胞周期等。
使用说明
1. 提前将消化酶从-20℃冰箱取出置于2-8℃融化;
2. 将本产品按照1: 1比例稀释组织消化酶 (货号O1521148),配制成工作浓度;
3. 将组织转移至培养皿中,加入3 mL组织清洗液 (货号O1521152)冲洗组织,反复2-3次;
4. 在无菌条件下将组织机械切割成1~2 mm³大小之后转移至15 mL无菌离心管中;
5. 加入5 mL组织清洗液,1500 rpm常温离心3 min;
6. 弃上清,加入12 mL工作浓度的消化酶,标记样本名称及编号并封口,放入恒温振荡培养箱中消化,设置参数37 ℃、200~300 rpm (根据样本质地设置转速),待组织消化呈团絮状,没有明显肉眼可见硬块终止消化;
7. 取出消化好的样本,1500 rpm室温离心3 min。
8. 弃上清后,加入6 mL含血清培养基进行终止消化,1500 rpm室温离心3 min。
9. 弃上清后加入原代细胞培养基重悬后待用。
注意事项
1. 使用产品时应注意无菌操作,避免污染。
2. 本产品配套组织消化酶使用,使用时按照1: 1稀释。
3. 请于保质期内使用本产品,超出保质期后,将无法达到理想的使用效果。
4. 本产品仅用于科研用途,不能用于体外诊断。
This product is applied in conjunction with Tissue Digestion Enzyme (Catalog No. O1521148). It contains a variety of nutritional factors, which can effectively prevent excessive tissue damage during digestion and improve the viability of primary cells isolated from digested tissues. The resulting cell suspension is suitable for primary cell culture and multiple flow cytometry-based cell biology assays, including apoptosis, cell proliferation, cell cycle analysis and other related experiments.
Instructions for Use
1. Take the digestion enzyme out of the -20℃ refrigerator in advance and thaw it at 2–8℃.
2. Dilute Tissue Digestion Enzyme (Catalog No. O1521148) with this product at a 1:1 ratio to prepare working solution.
3. Transfer tissues into a petri dish, add 3 mL Tissue Washing Solution (Catalog No. O1521152) to rinse tissues, and repeat rinsing 2–3 times.
4. Under aseptic conditions, mechanically cut tissues into 1–2 mm³ pieces, then transfer them into setrile 15 mL centrifuge tubes.
5. Add 5 mL Tissue Washing Solution and centrifuge at 1500 rpm at room temperature for 3 minutes.
6. Discard the supernatant, add 12 mL working-concentration digestion enzyme. Label sample names and numbers, seal the tubes, and place them in a constant temperature shaking incubator for digestion. Set parameters at 37℃, 200–300 rpm (adjust rotational speed according to tissue texture). Terminate digestion when tissues become flocculent with no visible hard masses.
7. Take out digested samples and centrifuge at 1500 rpm at room temperature for 3 minutes.
8. Discard the supernatant, add 6 mL serum-containing medium to stop digestion, and centrifuge at 1500 rpm at room temperature for 3 minutes.
9. Discard the supernatant, resuspend cells with primary cell medium for later use.
Precautions
1. Strict aseptic operation must be followed to avoid contamination.
2. This product is exclusively matched with tissue digestion enzyme and diluted at 1:1 ratio before use.
3. Use the product within the valid shelf life. Desired experimental performance cannot be ensured after expiration.
4. For research use only, not for in vitro diagnostic applications.