是否无菌：是
小分子化合物转染试剂是一款优秀、低成本、值得信赖的瞬时性转染试剂。在HEK293和CHO表达系统中，可在大的生产容器内（从96孔板到100L生物反应器）提供连续性的高基因表达。本品是无菌溶液，浓度为1mg/ml，可直接使用。按照小分子化合物：转染试剂=1：3的比例来操作，1ml 本品足以用来转染330μg小分子化合物。
使用说明：（贴壁细胞）
1. 铺板：
转染前18-24h进行铺板，调整合适的细胞密度（参考表1），使其在转染时细胞密度达60-80%。
2. 转染：（以6孔板为例）
a. 转染前1-2h，每孔替换为3ml含2%血清的新鲜生长培养基，转染前去除该培养基。
b. 制备小分子化合物转染试剂-小分子化合物转染复合物（请严格按照以下步骤进行）：
①往300μl无血清培养基加入2μg质粒小分子化合物，混匀。
②在混合物内加入6μl 小分子化合物转染试剂（1mg/ml）（小分子化合物：小分子化合物转染试剂=1：3），混匀。
③无菌环境，室温静置30min以形成小分子化合物转染试剂-小分子化合物转染复合物。
④用移液枪上下吹打3次，轻轻混匀。
c. 将小分子化合物转染试剂-小分子化合物转染复合物转到孔内。轻轻晃动培养皿或轻微涡旋，使复合物分散均匀。
3. 孵育：
a. 37°C，5% CO₂培养箱内培养细胞，转染12-18h，去除含小分子化合物转染试剂-小分子化合物复合物的培养液，更换新鲜的培养基。
b. 通常，转染36-48h后能检测到重组蛋白表达。一般在转染后72-96h能观察到最大水平的表达。
注意事项：
1. 收到本品或第一次使用前，请根据单次用量分装后置于-20°C冻存，尽量减少反复冻融次数。
2. 本品为无菌溶液，请严格按照无菌操作进行。
3. 请使用高质量的无内毒素质粒。通过260nm光吸收测定小分子化合物浓度，260nm/280nm比值确定小分子化合物纯度（比值应该在1.8～2.0的范围之内）。如有条件，请通过琼脂糖凝胶电泳检测质粒的完整性。
4. 请使用保存适当和经常传代的健康细胞。确保培养基没有被细菌、真菌或支原体污染。细胞如果是近期复苏的液氮冻存细胞，请在转染前至少传代两次。
5. 对于某些类型的细胞如HEK-293、HEK293T、NIH/3T3和COS细胞，在转染前两天铺板可显著提高重组蛋白的表达水平。如果选择转染前两天铺板，可适当降低铺板密度，以确保转染时细胞的汇合度仍为60-80%。
6. 对于接触抑制敏感的细胞，可适当降低铺板密度。
7. 推荐用商业化的无血清培养基比如Optimal-MEM Reduced Serum Medium (with Phenol Red) (O778393) 来稀释小分子化合物和小分子化合物转染试剂，制备转染复合物。
8. 转染过程中请不要使用抗生素。
9.本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
10.为了您的安全和健康，请穿实验服并戴一次性手套操作。
The Small Molecule Compound Transfection Reagent is an excellent, low-cost, and reliable transient transfection reagent. In HEK293 and CHO expression systems, it enables consistent high gene expression in large-scale production vessels (ranging from 96-well plates to 100L bioreactors). This product is a sterile solution with a concentration of 1 mg/mL and can be used directly. The recommended operating ratio is small molecule compound : transfection reagent = 1 : 3; 1 mL of this product is sufficient for transfecting 330 μg of small molecule compounds.
Instructions for Use: (Adherent Cells)
1. Plating:
Perform cell plating 18–24 hours before transfection. Adjust to an appropriate cell density (refer to Table 1) to ensure the cell density reaches 60–80% at the time of transfection.
2. Transfection: (Taking 6-well plates as an example)
a. 1–2 hours before transfection, replace the medium in each well with 3 mL of fresh growth medium containing 2% serum. Remove this medium before transfection.
b. Prepare the small molecule compound transfection reagent-small molecule compound transfection complex (follow the steps below strictly):
① Add 2 μg of plasmid small molecule compound to 300 μL of serum-free medium and mix well.
② Add 6 μL of small molecule compound transfection reagent (1 mg/mL) to the mixture (ratio of small molecule compound : small molecule compound transfection reagent = 1 : 3) and mix well.
③ Leave the mixture to stand at room temperature for 30 minutes in a sterile environment to form the small molecule compound transfection reagent-small molecule compound transfection complex.
④ Pipette up and down 3 times to mix gently.
c. Transfer the small molecule compound transfection reagent-small molecule compound transfection complex into the wells. Gently rock the culture dish or vortex slightly to distribute the complex evenly.
3. Incubation:
a. Incubate the cells in a 37°C, 5% CO₂ incubator. After 12–18 hours of transfection, remove the culture medium containing the small molecule compound transfection reagent-small molecule compound complex and replace it with fresh medium.
b. Typically, recombinant protein expression can be detected 36–48 hours after transfection. The maximum level of expression is generally observed 72–96 hours after transfection.
Precautions:
1. Upon receipt of this product or before first use, aliquot it according to single-use volumes and store at -20°C. Minimize the number of freeze-thaw cycles as much as possible.
2. This product is a sterile solution; strictly follow aseptic techniques during operation.
3. Use high-quality endotoxin-free plasmids. Determine the concentration of small molecule compounds by measuring light absorption at 260 nm, and assess the purity of small molecule compounds using the 260 nm/280 nm absorbance ratio (the ratio should be in the range of 1.8–2.0). If conditions permit, verify the integrity of plasmids via agarose gel electrophoresis.
4. Use healthy cells that are properly preserved and regularly passaged. Ensure the medium is free from contamination by bacteria, fungi, or mycoplasma. For cells recently recovered from cryopreservation in liquid nitrogen, passage them at least twice before transfection.
5. For certain cell types such as HEK-293, HEK293T, NIH/3T3, and COS cells, plating the cells two days before transfection can significantly improve the expression level of recombinant proteins. If you choose to plate the cells two days before transfection, appropriately reduce the plating density to ensure the cell confluency remains 60–80% at the time of transfection.
6. For cells sensitive to contact inhibition, appropriately reduce the plating density.
7. It is recommended to use commercial serum-free media (e.g., Optimal-MEM Reduced Serum Medium (with Phenol Red) (O778393)) to dilute small molecule compounds and the small molecule compound transfection reagent for preparing transfection complexes.
8. Do not use antibiotics during the transfection process.
9. This product is intended solely for scientific research by professional personnel. It shall not be used for clinical diagnosis or treatment, nor for food or pharmaceutical purposes, and shall not be stored in ordinary residential premises.
10. For your safety and health, wear a lab coat and disposable gloves during operation.
ZJ25F1230682
危险品化学品经营许可证（带存储）