本产品为白色或淡黄色速溶颗粒,每袋可配制1 L 1×免冰浴快速转膜缓冲液,15~40 min即可完成Western Blot湿转法转膜。配制过程中,可用无水乙醇替代甲醇,转膜全程产热量小,无需冰浴。
操作步骤:
1. 将大约600 mL蒸馏水倒入烧杯,并向其中缓慢加入1袋免冰浴快速转膜缓冲液速溶颗粒,搅拌溶液直至颗粒完全溶解。
2. 向步骤1所得溶液中加入蒸馏水,定容至900 mL。
3. 向步骤2所得溶液中加入100 mL无水乙醇或甲醇,混匀即为1×湿转法免冰浴快速转膜缓冲液。
转膜条件参考:
1. 梯度胶,浓度4%~15%,厚度1.5 mm,彩色预染蛋白分子量标准 (10-250kDa) rp192041,恒流400 mA,30 min,所有条带即可完成转膜。
2. 普通PAGE胶,转膜效率与胶浓度有关,对于各浓度1.0 mm凝胶,采用恒流400 mA,按如下时长即可将150 kDa以下蛋白转印完全,若蛋白分子量大于150 kDa,需延长转膜时间5~10 min:
胶浓度
| 6%
| 7.5%
| 10%
| 12.5%
|
推荐时长
| 15 min
| 20 min
| 30 min
| 35 min
|
3. PAGE胶厚度越大,转膜效率越低,对于厚度1.5 mm的凝胶,可将条目2中转膜时间相应延长5~10 min。
注意事项:
1. 为了您的安全和健康,请穿实验服并戴一次性手套操作。
2. 本产品仅限科研使用。
This product is a white or pale yellow instant granular powder. Each bag can be prepared into 1 L of 1× ice-bath-free rapid transfer buffer, enabling the completion of Western Blot wet transfer in 15–40 minutes. During preparation, absolute ethanol can be used as a substitute for methanol. The entire transfer process generates little heat, eliminating the need for ice bathing.
Operating Steps:
1. Pour approximately 600 mL of distilled water into a beaker, slowly add 1 bag of ice-bath-free rapid transfer buffer instant granular powder to it, and stir the solution until the granules are completely dissolved.
2. Add distilled water to the solution obtained in Step 1 and dilute to a final volume of 900 mL.
3. Add 100 mL of absolute ethanol or methanol to the solution obtained in Step 2, and mix well to obtain the 1× ice-bath-free rapid transfer buffer for wet transfer.
Reference for Transfer Conditions:
1. For gradient gels (concentration: 4%~15%, thickness: 1.5 mm) with pre-stained protein molecular weight marker (10-250 kDa, Cat. No.: rp192041), transfer all bands completely under a constant current of 400 mA for 30 minutes.
2. For regular PAGE gels, transfer efficiency is related to gel concentration. For 1.0 mm-thick gels of various concentrations, proteins with molecular weight below 150 kDa can be completely transferred at a constant current of 400 mA for the recommended durations listed below. If the protein molecular weight exceeds 150 kDa, extend the transfer time by 5~10 minutes:
Gel Concentration
| 6%
| 7.5%
| 10%
| 12.5%
|
Recommended Duration
| 15 min
| 20 min
| 30 min
| 35 min
|
3. The thicker the PAGE gel, the lower the transfer efficiency. For 1.5 mm-thick gels, extend the transfer time specified in Item 2 by 5~10 minutes accordingly.
Precautions:
1. For your safety and health, please wear a lab coat and disposable gloves during operation.
2. This product is for research use only.