Product Introduction
|
Product Name |
Cat. No. |
Spec. |
|
5×Tris-Glycine Transfer Buffer |
G2163-1L |
1 L |
Product Description/Introduction
This product is a 5x concentrated solution, with the main components of 100 mM Tris and 0.96 M glycine, without methanol or absolute ethanol. Before use, take 200 mL of this product and add 500 mL of deionized water or distilled water, mix well, then add 200 mL of absolute ethanol, mix well, and finally add deionized water or distilled water to a final volume of 1 L. After thorough mixing, a 1x Tris-Glycine transfer buffer with 20 mM Tris, 192 mM glycine, 20% absolute ethanol, and pH 8.2-8.6 at 25°C is obtained. It is suitable for wet or semi-dry transfer in Western blot experiments.
Storage and Shipping Conditions
Ship and store at room temperature; Valid for 12 months.
Assay Protocol/Procedures
1. For regular small to medium molecular proteins (25-70 kDa), it is recommended to use a transfer condition of 300 mA for approximately 30 minutes.
2. For regular large molecular proteins, the transfer time can be extended as needed, and it is recommended to use a transfer condition of 300 mA for 45-60 minutes.
3. For small molecular proteins below 20 kDa, it is recommended to use a smaller pore size PVDF membrane of 0.22 μm. The transfer conditions can be 200 mA for 20-25 minutes or 300 mA for 15-20 minutes. Alternatively, the methanol content (can be replaced with ethanol) can be increased to 25% to enhance the fixation of small molecular proteins from the gel.
4. For proteins with particularly large molecular weights, the transfer time can be further extended. It is recommended to use a transfer condition of 300 mA for 60-90 minutes or longer. Additionally, SDS can be added to the transfer buffer to a final concentration of approximately 0.025-0.1% to enhance the release of proteins from the gel. Due to the longer transfer time, attention should be paid to the cooling effect of the cooling module, and if necessary, a new cooling module can be replaced midway.
Note
1. Please use the working solution as soon as possible after preparation. It is recommended to dilute the reagent with laboratory purified water (G4701-500ML).
2. Crystallization may occur at lower environmental temperatures, which is a normal phenomenon. It can be heated at 37°C, during which time it is mixed upside down several times, and then diluted for use after it has completely dissolved.
3. The normal color of this product is colorless and transparent, if it appears light brown or yellowish brown, it should be discarded promptly.
4. Wear lab coat and disposable gloves when handling.
For Research Use Only!
