SDS-PAGE loading buffer,4×(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 bind with protein to produce a large amount of negative charge on the protein-SDS complex, then the charge of the protein itself is completely covered by SDS, eliminating the difference in the charge of various proteins. SDS can also break 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 10 microliters of sample buffer per 30 microliters of protein sample (4 times dilution) to use. If the protein sample 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. After cooling to room temperature, centrifuge at 10,000-14000rpm for 2-5 minutes, take the supernatant directly onto the sample for electrophoresis.

Note:

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

2, this reagent contains DTT, there is a certain toxicity, for your safety and health, please wear a laboratory coat and wear disposable gloves operation.

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.