是否无菌：是
通用细胞转染试剂是一款基于聚合物的同时适用于贴壁细胞、悬浮细胞和原代细胞转染的试剂，能够有效地将DNA、mRNA、siRNA等核酸转入多种细胞株包括Expi293、CHO-S、K562、THP-1等悬浮细胞系，HEK-293T、4T1、A549、PC等贴壁细胞系，眼部和神经原代细胞。该产品具有操作简单、蛋白表达量高、细胞毒性低、细胞通用性好等优势。产品以无菌的液体形式提供。
产品组分及储存条件：
T751608
| Components
| 100μL
| 500μL
| 1.5mL
| Storage
|
T751608A
| 通用细胞转染试剂
| 100μL
| 500μL
| 1.5mL
| -20℃
|
T751608B
| Transfection Buffer
| 2mL
| 10mL
| 30mL
| -20℃ |
实验材料：
• 本产品包含通用细胞转染试剂和 Transfection Buffer
• ⾼纯度的核酸储液（0.5∼5 μg/μL）
• ⽆⾎清培养基（如 O778393 Optimal-MEM减血清培养基 (含酚红) ）
质量控制：
• -20℃保存条件下有效期⼀年
• 本产品经严格的质量检验证明，⽆⽣物污染
注意事项：
• 常温下制备通用细胞转染试剂-核酸复合物，孵育时间15min。转染时使用无血清培养基（如O778393 Optimal-MEM减血清培养基 (含酚红))和灭酶管。
• 利用本产品转染4-6h后，补加一体积的新鲜完全培养基。
• 初次使用应该优化核酸浓度与通用细胞转染试剂的比例以得到最高的转染效率。测试使用推荐范围内的产品用量。
• 转染时请使用适当保存和经常传代的健康细胞，并确保培养基无细菌、真菌或支原体污染。如细胞是近期复苏的液氮冻存细胞，请在转染前至少传代两次。
• 本产品应在-20°C密闭保存，使用前，提前10min置于室温下融化。
• 转染试剂使用后请立即盖好盖子，并置于-20°C密封保存。
转染效果示例：
图1. 通用细胞转染试剂和竞品在Exipi293、CHO-S悬浮细胞中递送pDNA48h后的细胞阳性率和绿色荧光蛋白表达量，在GFP-HeLa中递送GFP-siRNA 48h后的基因沉默效率(24孔板，每孔转染1μg pDNA)。
图2. 通用细胞转染试剂和竞品在GFP-HeLa中递送GFP-siRNA 48h后的基因沉默效率。(24孔板，每孔转染1.5 μg siRNA；GFP荧光越暗，表明GFP表达量越低、沉默效率越高)。
图3. 通用细胞转染试剂和竞品在293T、4T1等贴壁细胞中递送pDNA 48h后的荧光表达(24孔板，每孔转染1μg pDNA)。
图4. 通用细胞转染试剂和竞品在293T、DC等贴壁细胞中递送mRNA 48h后的荧光表达(24孔板，每孔转染1μg mRNA)。
按照下表转染细胞。每种反应混合物体积为单个孔的体积，且考虑了移液差异。按比例计算其他孔的体积。
放附件 [T751608] 通用细胞转染试剂-中英文步骤表格中，中文表格
[T751608] 通用细胞转染试剂-中英文步骤表格.pdf
通用细胞转染试剂----转染步骤详细说明（以24孔板为例）
1. 在使用前，请提前10min 将通用细胞转染试剂置于室温下融化。
2. 取20μL Transfection Buffer （pH~5）于1.5mL 离心管中，并向其中加入1μg 核酸，得到核酸稀释液。
3. 取1μL 通用细胞转染试剂于另一1.5mL 离心管中（通用细胞转染试剂的用量可在0.7–1.5μL范围内进行优化，以寻求最佳转染效果）。
4. 将步骤2中的核酸稀释液转移至步骤3中含通用细胞转染试剂的离心管中，并充分混合均匀，室温下孵育15 min。
5. 将孵育好的通用细胞转染试剂–核酸复合物与480μL 无血清培养基 (如O778393 Optimal-MEM减血清培养基 (含酚红))混合后加至细胞；6.37°C孵育细胞4-6h 后补加一体积的完全培养基，继续37°C孵育细胞2-4天后分析转染细胞。
通用细胞转染试剂----转染悬浮细胞步骤详细说明 （以125mL培养瓶、30mL培养体系为例）
1. 在使用前，请提前10min 将通用细胞转染试剂置于室温下融化。
2. 取1.2mL Transfection Buffer （pH~5）于1.5mL 离心管中，并向其中加入60μg 核酸，得到核酸稀释液。
3. 取60μL 通用细胞转染试剂于另一1.5mL 离心管中（通用细胞转染试剂的用量可在32–90μL范围内进行优化，以寻求最佳转染效果）。
4. 将步骤2中的核酸稀释液转移至步骤3中含通用细胞转染试剂的离心管中，并充分混合均匀，室温下孵育15 min。
5. 将孵育好的通用细胞转染试剂-核酸复合物直接加入到细胞培养瓶中，37°C孵育细胞2-4天后，分析细胞转染情况。
The Universal Cell Transfection Reagent is a polymer-based reagent suitable for transfecting adherent cells, suspension cells, and primary cells. It can effectively deliver nucleic acids such as DNA, mRNA, and siRNA into a variety of cell lines, including suspension cell lines like Expi293, CHO-S, K562, and THP-1, as well as adherent cell lines such as HEK-293T, 4T1, A549, and PC, along with ocular and neural primary cells. This product has advantages including simple operation, high protein expression level, low cytotoxicity, and good cell universality. It is supplied as a sterile liquid.
Product Components and Storage Conditions:
T751608
| Components
| 100μL
| 500μL
| 1.5mL
| Storage
|
T751608A
| Universal Cell Transfection Reagent
| 100μL
| 500μL
| 1.5mL
| -20℃
|
T751608B
| Transfection Buffer
| 2mL
| 10mL
| 30mL
| -20℃
|
Experimental Materials:
• This product contains the Universal Cell Transfection Reagent and the Transfection Buffer .
• High-purity nucleic acid stock solution (0.5–5 μg/μL).
• Serum-free medium (e.g., O778393 Optimal-MEM Reduced Serum Medium (with Phenol Red) ).
Quality Control:
• The shelf life is 1 year when stored at -20°C.
• Strict quality inspection has confirmed that this product is free from biological contamination.
Precautions:
• Prepare the Universal Cell Transfection Reagent-nucleic acid complex at room temperature, with an incubation time of 15 minutes. For transfection, use serum-free medium (e.g., O778393 Optimal-MEM Reduced Serum Medium (with Phenol Red)) and enzyme-free tubes.
• After transfecting with this product for 4-6 hours, add an equal volume of fresh complete medium.
• For first-time use, the ratio of nucleic acid concentration to the Universal Cell Transfection Reagent should be optimized to achieve the highest transfection efficiency. Test using the product dosage within the recommended range.
• For transfection, use healthy cells that are properly preserved and regularly passaged, and ensure the medium is free from bacterial, fungal, or mycoplasma contamination. If the cells are recently revived from liquid nitrogen-frozen stocks, they should be passaged at least twice before transfection.
• This product should be stored hermetically at -20°C. Before use, place it at room temperature to thaw for 10 minutes in advance.
• After using the transfection reagent, immediately replace the cap tightly and store it hermetically at -20°C.
Example of transfection effect:
Figure 1. Cell positive rate and green fluorescent protein expression level 48 hours after delivering pDNA in Exipi293 and CHO-S suspension cells using the Universal Cell Transfection Reagent and competing products, as well as gene silencing efficiency 48 hours after delivering GFP-siRNA in GFP-HeLa cells (24-well plate, 1 μg of pDNA transfected per well).
Figure 2. Gene silencing efficiency 48 hours after delivering GFP-siRNA in GFP-HeLa cells using the Universal Cell Transfection Reagent and competing products. (24-well plate, 1.5 μg of siRNA transfected per well; the dimmer the GFP fluorescence, the lower the GFP expression level and the higher the silencing efficiency.)
Figure 3. Fluorescence expression 48 hours after delivering pDNA in adherent cells such as 293T and 4T1 using the Universal Cell Transfection Reagent and competing products (24-well plate, 1 μg of pDNA transfected per well).
Figure 4. Fluorescence expression 48 hours after delivering mRNA in adherent cells such as 293T and DC using the Universal Cell Transfection Reagent and competing products (24-well plate, 1μg of mRNA transfected per well).
Transfect cells according to the table below. The volume of each reaction mixture refers to the volume for a single well, with pipetting differences taken into account. Calculate the volumes for other wells in proportion.
[T751608] Universal Cell Transfection Reagent.pdf
Universal Cell Transfection Reagent----Detailed Instructions for Transfection Steps (Taking 24-well Plate as an Example)
1. Before use, place the Universal Cell Transfection Reagent at room temperature to thaw 10 minutes in advance.
2. Take 20 μL of Transfection Buffer (pH ~5) into a 1.5 mL centrifuge tube, and add 1 μg of nucleic acid to it to obtain the nucleic acid dilution.
3. Take 1 μL of Universal Cell Transfection Reagent into another 1.5 mL centrifuge tube (The dosage of Universal Cell Transfection Reagent can be optimized within the range of 0.7–1.5 μL to achieve the best transfection effect).
4. Transfer the nucleic acid dilution from Step 2 into the centrifuge tube containing Universal Cell Transfection Reagent from Step 3, mix thoroughly until uniform, and incubate at room temperature for 15 minutes.
5. Mix the incubated Universal Cell Transfection Reagent-nucleic acid complex with 480 μL of serum-free medium (e.g., O778393 Optimal-MEM Reduced Serum Medium (with Phenol Red)), then add the mixture to the cells.
6. After incubating the cells at 37°C for 4–6 hours, add an equal volume of complete medium. Continue incubating the cells at 37°C for 2–4 days, then analyze the transfected cells.
Universal Cell Transfection Reagent----Detailed Instructions for Transfecting Suspension Cells (Taking a 125mL Culture Flask and 30mL Culture System as Examples)
1. Before use, place the Universal Cell Transfection Reagent at room temperature to thaw 10 minutes in advance.
2. Take 1.2 mL of Transfection Buffer (pH ~5) into a 1.5 mL centrifuge tube, and add 60 μg of nucleic acid to it to obtain the nucleic acid dilution.
3. Take 60 μL of Universal Cell Transfection Reagent into another 1.5 mL centrifuge tube (The dosage of Universal Cell Transfection Reagent can be optimized within the range of 32–90 μL to achieve the best transfection effect).
4. Transfer the nucleic acid dilution from Step 2 into the centrifuge tube containing the Universal Cell Transfection Reagent from Step 3, mix thoroughly until uniform, and incubate at room temperature for 15 minutes.
5. Directly add the incubated Universal Cell Transfection Reagent-nucleic acid complex into the cell culture flask, incubate the cells at 37°C for 2–4 days, and then analyze the cell transfection status.
ZJ26F0131401
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