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.
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