Description
Product Description
This product consists of white or pale yellow instant-dissolving granules; each sachet yields 1 L of 1× rapid transfer buffer that eliminates the need for an ice bath, enabling Western Blot wet transfer to be completed in 15–40 minutes. Anhydrous ethanol can be substituted for methanol during preparation, and minimal heat is generated throughout the transfer process, obviating the need for an ice bath.
Operating Steps
1. Pour approximately 600 mL of distilled water into a beaker, slowly add one packet of rapid transfer buffer granules (designed for use without an ice bath), and stir the solution until the granules are completely dissolved；
2. Add distilled water to the solution obtained in Step 1 and adjust the volume to 900 mL;
3. Add 100 mL of anhydrous ethanol or methanol to the solution obtained in Step 2 and mix well to prepare the 1× rapid transfer buffer for wet transfer (ice-bath-free).
Reference conditions for membrane transfer
1. Gradient gel (4%–15%, 1.5 mm thick), three-color prestained molecular weight marker WJ103 (10–250 kDa); transfer performed at a constant current of 400 mA for 30 minutes, resulting in the transfer of all bands.
For standard PAGE gels, transfer efficiency depends on gel concentration. Using a constant current of 400 mA, proteins smaller than 150 kDa can be completely transferred from 1.0 mm gels of various concentrations within the durations listed below; for proteins larger than 150 kDa, the transfer time should be extended by 5–10 minutes:
|
Gel concentration |
6% |
7.5% |
10% |
12.5% |
|
Recommended duration |
15 min |
20 min |
30 min |
35 min |
3. The greater the PAGE gel thickness, the lower the transfer efficiency; for 1.5 mm thick gels, the transfer time specified in Item 2 can be extended by 5–10 minutes.
Precautions
1. For your safety and health, please wear a lab coat and disposable gloves while performing operations.
2. This product is for research use only.