Introduction:
Western transfer buffer is used to transfer proteins from polyacrylamide gels to membranes for western blotting.
The main components of this product are Tris and glycine, no methanol. It is mainly used for protein transfer to PVDF membrane or nitrocellulose membrane, not suitable for protein transfer to nylon membrane. This product is a concentrated solution, which needs to be diluted to 1× working concentration before use.
Storage: Store at 4℃, valid for 3 months.
Protocol (for reference only):
- 1× western transfer buffer can be used for membrane transfer directly. 5× and 10× western transfer buffer should be diluted to 1× working concentration before use.
- Dilute the 5× western transfer buffer with the ratio of 5×transfer buffer: methanol: deionized water =1: 1: 3 and mix well to obtain 1× transfer buffer.
- Dilute the 10× western transfer buffer with the ratio of 10×transfer buffer: methanol: deionized water =1: 2: 7 and mix well to obtain 1× transfer buffer.
- Perform protein transfer at 4°C to prevent protein degradation.
Notes:
- Keep it tightly closed and use it up as soon as possible after opening.
- Choose high-grade methanol, analytically pure methanol can also be used.
- This product does not contain SDS. When the molecular weight of the target protein is large or the hydrophobicity is strong, a small amount of SDS (the reference working concentration is 0.025-0.1%) can be appropriately added to increase the transfer effect.
- For your safety and health, please wear personal protective equipment and clothing when operating.
- For research use only. Not for use in diagnostic procedures.