
Western Rapid Transfer Buffer (10X)
$15.00 - $160.00
All products have special prices for bulk purchase, please contact for more details if required.
Cat. No.: WRTB-100 (for 100ml)
Cat. No.: WRTB-500 (for 500ml)
Cat. No.: WRTB-2k (for 4×500ml)
Description
Western Rapid Transfer Buffer (10X) is a 10-fold concentrated, safe, and non-toxic buffer used for rapid wet transfer in Western blotting.
This product is safe and non-toxic. It is a 10-fold concentrated (10X) solution that needs to be diluted to 1X with appropriate water and ethanol before use. It does not contain any toxic or harmful components, and there is no need to use toxic reagents such as methanol.
This product enables rapid transfer. When using this rapid transfer buffer, protein transfer from polyacrylamide gels to membranes such as PVDF or nitrocellulose membranes (NC membranes) can be achieved in 15-20 minutes at a constant current of 500mA (external ice bath is recommended) or approximately 20-25 minutes at 400mA constant current (internal ice box can be used without an external ice bath). It generates less heat and provides good transfer efficiency. Under the same conditions, traditional transfer buffers are more prone to excessive heating, and they require longer transfer times compared to this rapid transfer buffer.
The transfer efficiency of this rapid transfer buffer is comparable to or slightly better than traditional transfer buffers. For a comparison of the pre- and post-transfer effects between this rapid transfer buffer and traditional transfer buffers. In the figure, it can be observed that a 270 kDa protein molecular weight marker was completely transferred with a current of 400mA for 25 minutes. If the target protein has a smaller molecular weight, the transfer time can be shortened to 20 or 15 minutes accordingly, or an external ice bath with 500mA can be considered for 15-20 minutes of transfer.
This rapid transfer buffer generates significantly less heat compared to traditional transfer buffers. When conducting transfer at a constant current of 400mA, there is no need for an external ice bath in the outer chamber; using only the internal ice box of the transfer apparatus is sufficient to prevent overheating. If a faster transfer is desired, 500mA constant current can be used, and in this case, an external ice bath is recommended.
The rapid transfer buffer is convenient to use and does not require pH adjustment.
This product is suitable for the transfer of SDS-PAGE gels prepared with manually prepared Tris-Gly and Hepes buffer systems, as well as precast gels prepared with Tris-Gly, Hepes, and Bis-Tris buffer systems.
The transfer buffer can be reused after use. It can be recycled and reused for 1-2 additional transfers.
Precautions
- If the color of the transfer buffer turns light brown or yellow-brown, it should be discarded.
- If the transfer buffer has been stored for a long time at 4°C and there is precipitation, it can be placed in a water bath and fully dissolved before use.
- It is recommended to use analytical grade ethanol or higher purity ethanol.
- If a higher current or voltage is set during the transfer, there may be significant heat generation, known as Joule heating effect, which can decrease the resistance of the transfer buffer. This can ultimately affect the transfer efficiency, buffering capacity of the transfer buffer, and may even cause gel melting or adhesion to the membrane due to overheating. Since the use of this product generates less heat, when performing a 400mA constant current transfer, cooling using only the built-in icebox in the transfer tank is sufficient. For better results, the entire transfer tank can be placed in an ice bath for cooling. It is also advisable to pre-cool the transfer buffer at 4°C or in an ice bath before use.
- When using this product for transfer, if a constant current of 400mA is set, the voltage typically starts at around 200V, regardless of whether 1 or 2 gels are being transferred in the same transfer tank. During the transfer, as the temperature of the transfer buffer gradually increases and the resistance decreases, the voltage usually decreases to 100-150V at the end. Transfer efficiency and electric field strength (represented by E) are related to the electric charge carried by the proteins, and electric field strength is mainly determined by the voltage and the distance between the electrodes, i.e., E=U/d.
- Due to differences in brand and model of power supplies used for transfer, there can be significant variations in the maximum current and power. Usually, high-power power supplies are relatively more expensive but more suitable for transfer. For example, the Bio-Rad Powerpac Basic Power Supply has a maximum power of 75W, and when a constant current of 400mA is set, exceeding 187V will exceed the maximum power of the power supply, triggering overload protection. Some power supplies automatically lower the voltage or current, while others display an error and stop the transfer. In cases where the transfer stops, it is necessary to reduce the current or voltage or choose a power supply with a higher power.
- During transfer, factors such as the temperature of the transfer buffer, gel parameters (e.g., quantity, thickness, ion concentration, acrylamide concentration), thickness of the filter paper, number of transfer tanks used simultaneously, ion concentration of the transfer buffer, volume of the transfer buffer, and number of times the transfer buffer has been used may all affect the current and voltage. If multiple transfer tanks need to be used simultaneously, it is recommended to use a high-current, high-power power supply or set a lower current or voltage for a longer transfer time.
- If an error occurs and the transfer stops, it is necessary to investigate whether it is due to voltage or current overload or other abnormalities such as excessive or insufficient transfer buffer, short circuit or open circuit in the transfer tank.
- This product is intended for scientific research use by professionals only and should not be used for clinical diagnosis or treatment, or for food or drug purposes. It should not be stored in regular residential areas.
- For your safety and health, please wear appropriate lab attire and disposable gloves when handling.
Storage
Store at 4°C, effective for one year. Store at room temperature, effective for at least one week.
Only for research and not intended for treatment of humans or animals
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