Product Introduction
|
Product Name |
Cat. No. |
Spec. |
|
5×Ice-Bath Free Fast Transfer Buffer |
G2164-1L |
1 L |
Product Description/Introduction
This product is a 5x concentratedIce-Bath Free Fast Transfer Buffer, it is a safe and harmless buffer for rapid wet transfer in Western Blot, which can efficiently and quickly transfer proteins to thetransfer membrane. Without methanol, thetransfer process can be completed within 15-30 min.
Safe and non-toxic: This product does not contain toxic and harmful components, and does not need to use highly toxic reagents such as methanol.
The transfer effect of the transfer buffer is the same or better than that of theconventional transfer buffer.
Rapidtransfer without ice bath: With rapid transfer buffer, proteins on gels can be transferred to imprinted membranes such as PVDF membrane or nitrate fiber membrane (NC membrane) in a constant flow membrane of 400 mA for 15-30 min, and the heat production is lower than that of conventional transfer buffer.
Storage andShipping Conditions
Ship and store at room temperature; Valid for 12 months.
Assay Protocol/Procedures
1. Take 200 mL of concentrated ice-bath free fast transfer buffer and add 500 mL of deionized water or distilled water, mix well. Then add 200 mL of anhydrous ethanol, mix well. Finally, add deionized water or distilled water to a final volume of 1 L, mix well to obtain a 1× working solution.
2. Prepare the sandwich structure required for transfer, in the order from the negative electrode (black side) to the positive electrode (red side): sponge, filter paper, gel, transfer membrane, filter paper, sponge. Transfer the sandwich structure to the transfer tank, add the ice-bathfreefasttransfer buffer, set a constant current of 400 mA, and transfer the proteins for 15-30 minutes (this condition is for 1.0 mm thick gels, if the gel thickness is 0.75 mm, reduce the transfer time accordingly; if the gel thickness is 1.5 mm, extend the transfer time accordingly) to complete protein transfer.
|
Ice-Bath Free Fast Transfer Buffer |
constant current |
molecular weight size |
Recommended transfer time |
Corresponding initial voltage magnitude |
|
300 mA |
<20 kDa |
20 min |
~ 95 V |
|
|
20-100 kDa |
30 min |
|||
|
100-150 kDa |
50 min |
|||
|
>150 kDa |
>60 min |
|||
|
400 mA |
<20 kDa |
15 min |
~125 V |
|
|
20-100 kDa |
25 min |
|||
|
100-150 kDa |
45 min |
|||
|
>150 kDa |
>55 min |
Note
1. Please use the solution as soon as possible after preparation. It is recommended to dilute the reagent with laboratory pure water (G4701-500ML).
2. This product is suitable for wet transfer of protein gels in Tris-glycine, Bis-Tris and other systems.
3. The PVDF membrane should be pre-wetted with anhydrous ethanol for about 30 seconds before use.
4. Transfers within 30 minutes do not require an ice bath. For transfers exceeding 30 minutes, an ice bath is recommended.
5. Due to differences in the brand and model of the power supply used for transfer, there may be significant variations in the maximum current and power output. In some cases, when setting a constant current of 400 mA, overload protection may occur due to exceeding the maximum power. In this case, it is necessary to appropriately reduce the current or choose a power supply with higher power.
6. Factors such as the temperature of the transfer buffer, gel parameters (such as thickness, acrylamide concentration, ion concentration), filter paper thickness, volume of the transfer buffer, and number of times the transfer buffer has been used may affect the current and voltage during transfer. If overload occurs, please reduce the current appropriately and extend the transfer time.
7. The normal color of this product is colorless and transparent. If it appears light brown or yellowish-brown, it should be discarded promptly.
8. This product may experience crystal precipitation in low-temperature environments, which is normal. It can be moderately heated and dissolved with magnetic stirring before dilution and subsequent use.
For Research Use Only!
