Western快速转膜液 (20×, 湿转)是由阿拉丁自主研发的高效转膜液，可应用于湿转法转膜或者半干法转膜，能高效快速地将蛋白转移至PVDF膜或硝酸纤维素膜上，转移过程所产生的热量远远小于普通转膜缓冲液，从而减小对蛋白的损失。本产品安全无毒害，使用前仅需用水和乙醇稀释至1×（能在15-35分钟内快速完成转膜全过程）。
产品特性：
1.本产品转膜效率高，能在15-35分钟内快速完成转膜过程。
2.稳定性高，产热量小，减少对蛋白的损失。
3.兼容性好，对分子量跨度大的蛋白同样适用，有效解决大小蛋白不能在同一张膜上同时转移的问题，兼容Tris-Gly体系、Hepes体系、Bis-Tris体系等多种凝胶。
操作步骤：
1.1×快速转膜液使用前的配制方法：
如配制1L的1×快速转膜液请参考下表：
Western快速转膜液 (20X, 湿转)
| 50ml
|
无水乙醇
| 200ml
|
超纯水
| 750ml
|
准备5-6张裁剪好的滤纸和转印膜，如若使用PVDF膜需提前用无水甲醇浸泡至少10秒以上，再放在1×快速转膜液中。
2.操作步骤及转膜条件：
将滤纸和海绵浸泡于1×快速转膜液中，务必完全浸湿。
将凝胶在超纯水中漂洗2分钟，以去除胶表面的SDS；然后将凝胶浸泡于1×快速转膜液中。
请注意：漂洗时间切勿超过2分钟，否则分子量较大的蛋白无法完全转移。
3.请按照以下顺序完成转印三明治结构：
—>负极(阴极)
—>海绵
—>滤纸
—>凝胶
—>转印膜
—>滤纸
—>海绵(根据三明治的厚度选择是否使用)
—>正极(阳极)
将三明治结构转移至电泳槽中，设定恒流400mA，电泳时间15-35分钟完成蛋白转膜。
注意事项：
1.当转膜液长时间在2-8°C存放，若出现沉淀，可在水浴锅中温育并充分溶解后继续使用。
2.注意在使用乙醇时，不要求必须使用无水乙醇，如使用含水乙醇，请按照试剂中乙醇实际浓度计算加入量。
3.湿转转膜时电转仪外无需额外冰浴，即可确保无过热现象，如若希望更为快速地完成转膜，可使用400mA或更高的电流进行恒流转膜。
4.本品仅供科研使用，勿做其他用途。
5.为了您的安全和健康，请全程穿实验服并戴一次性手套操作。
Western Blot Rapid Transfer Buffer (20×, Wet Transfer) is a high-efficiency transfer buffer independently developed by Aladdin. It is suitable for both wet transfer and semi-dry transfer, and can transfer proteins to PVDF membranes or nitrocellulose membranes efficiently and rapidly. The heat generated during the transfer process is far lower than that of ordinary transfer buffers, thus reducing protein loss. This product is safe and non-toxic, and only needs to be diluted to 1× with water and ethanol before use (the entire transfer process can be completed quickly within 15-35 minutes).
Product Features
1. This product has high transfer efficiency and can complete the entire transfer process quickly within 15-35 minutes.
2. It features high stability and low heat generation, which reduces protein loss during the transfer.
3. It has excellent compatibility: applicable to proteins with a wide molecular weight range, effectively solving the problem that large and small proteins cannot be transferred onto the same membrane simultaneously. It is compatible with various gels such as Tris-Gly system, Hepes system and Bis-Tris system.
Operation Steps:
1. Preparation method of 1× Rapid Transfer Buffer before use:
For the preparation of 1L of 1× Rapid Transfer Buffer, refer to the table below:
Western Blot Rapid Transfer Buffer (20X, Wet Transfer)
| 50ml
|
Anhydrous Ethanol
| 200ml
|
Ultrapure Water
| 750ml
|
Prepare 5-6 pieces of cut filter paper and transfer membranes. If PVDF membranes are used, soak them in anhydrous methanol for at least 10 seconds in advance, then place them in 1× Rapid Transfer Buffer.
2. Operation Steps and Transfer Conditions:
Soak the filter paper and sponges in 1× Rapid Transfer Buffer, ensuring complete wetting.
Rinse the gel in ultrapure water for 2 minutes to remove SDS from the gel surface; then soak the gel in 1× Rapid Transfer Buffer.
Note: Do not rinse for more than 2 minutes, otherwise high-molecular-weight proteins cannot be transferred completely.
3. Assemble the Transfer Sandwich in the Following Order
→ Cathode
→ Sponge
→ Filter paper
→ Gel
→ Transfer membrane
→ Filter paper
→ Sponge (optional, based on the thickness of the sandwich)
→ Anode
Transfer the assembled sandwich to the electrophoresis tank, set a constant current of 400 mA, and run electrophoresis for 15–35 minutes to complete protein transfer.
Precautions:
1. If precipitation forms when the transfer buffer is stored at 2–8°C for an extended period, incubate it in a water bath and mix thoroughly to dissolve the precipitate before further use.
2. Anhydrous ethanol is not mandatory for use. If hydrous ethanol is used, calculate the added volume according to its actual concentration.
3. No additional ice bath is required outside the electroblotter for wet transfer, which ensures no overheating. For faster transfer, perform constant-current transfer at 400 mA or a higher current.
4. This product is for research use only (RUO) and not for any other purposes.
5. For your safety and health, wear a lab coat and disposable gloves for the entire operation process.
ZJ26F0938899
危险品化学品经营许可证（带存储）