SDS-PAGE loading buffer,2×(with DTT)

Product Introduction:

This product is suitable for SDS-PAGE(SDS denatured polyacrylamide gel electrophoresis) as a protein sample. Its main components are SDS, DTT, bromophenol blue, buffer salt solution and so on. SDS can combine with proteins to make the protein-SDS complex with a large amount of negative charge. At this time, the charge of the protein itself is completely covered by SDS, eliminating the difference in the charge of various proteins. SDS can also break the intramolecular and intermolecular hydrogen bonds and destroy the secondary and tertiary structures of protein molecules. DTT can break disulfide bonds between cysteine residues, destroying protein structures and eliminating differences between protein structures. Ultimately no charge and structural differences in the protein (subunit), electrophoresis speed is only related to the size of its molecular weight. Bromophenol blue is used as an indicator during electrophoresis, indicating roughly when the electrophoresis will end.

Instruction:

1. Please add 20 μl buffer for every 20 μl protein sample. If the protein concentration is too high, it can be diluted with double steaming water.

2. After mixing, heat in 100℃ water bath for 5-10 minutes to denature the protein.

3. Cool to room temperature, centrifuge for a few seconds, mix well and then centrifuge for 30 seconds, take the supernatant directly onto the sample.

Note:

1. When the concentration of polyacrylamide gel is 8%, the position of the bromophenol blue indicator band is about 30kd; when the gel concentration is 12%, it is about 20kd; when the gel concentration is 15%, it is about 10kd. Please judge the electrophoresis time according to your target strip.

2. This reagent contains DTT and has certain toxicity. For your safety and health, please wear a laboratory coat and disposable gloves.

3. The protein loading buffer contains bromophenol blue indicator, and the PH value is affected by the storage temperature. In the state of cryogenic storage, the solution may appear dark brown, which does not affect the use of the product.

Note：Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.

Remark：These protocols are for reference only. Solarbio does not independently validate these methods.

Note：

1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.

2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.

3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.