Cat.No.：D13021A
Price: ￥60
Specifications：
This product is a 5× concentrated loading buffer for protein sample preparation in SDS-PAGE (Sodium Dodecyl Sulfate - Polyacrylamide Gel Electrophoresis). It contains a tracer dye, bromophenol blue, which can be used for real-time monitoring of the electrophoresis process. When used, mix it with the protein sample to a 1× working concentration, and then incubate it in a water bath above 95℃ for 10 minutes to complete the denaturation treatment of the sample. It can achieve good component separation in gel electrophoresis and can be combined with staining methods such as Coomassie Brilliant Blue staining or silver staining, allowing the protein bands to be clearly developed.
The product formula has been optimized. Even after dilution to 1×, the density remains relatively high. During sample loading, it tends to sink. The indicator has good visibility. It can fully denature the protein samples within a short period of time, and the electrophoresis bands are clear and accurate.
| Cat. No. | Component | Specification | Quantity |
| B16021A | SDS-PAGE Loading Buffer(5×) | 1 mL | 5 |
-20℃.
This product is suitable for the pre-treatment and loading of protein samples in SDS-PAGE, and it should be used in combination with a hot water bath.
What is the cause of the precipitation in the sample buffer? How should it be dealt with?
SDS is prone to precipitate when cooled. Before use, heat the buffer solution at room temperature or in a water bath not exceeding 37℃ until the precipitation completely disappears, and then store it at room temperature immediately. Avoid prolonged exposure to high temperatures.
What are the usage ratios and heating conditions?
Mix the protein sample with the loading buffer at a ratio of 4:1 (that is, add 1 μL of 5× loading buffer for every 4 μL of protein sample), and mix well. Heat it in a boiling water bath at 100°C for 3–5 minutes to fully denature the protein. It is recommended to heat at 95°C for 5 minutes.
What should we do if the sample remains viscous after heating?
If the amount of cells or tissues is large and the genomic DNA content is high, after boiling, it may still be relatively viscous or have semi-transparent gel-like substances. The boiling time can be extended by 5-10 minutes, or an appropriate amount of sample buffer diluted to 1× can be added and then boiled for 3-5 minutes to break the genomic DNA and fully release the proteins. Alternatively, ultrasound or repeated aspiration with a 1 mL syringe can be used to break the genomic DNA.
Is the loading buffer suitable for non-denaturing PAGE?
Not applicable. This product contains SDS denaturing agent and is only suitable for SDS-PAGE denaturing gel electrophoresis, not for non-denaturing gel electrophoresis (Native-PAGE).
What are the differences between 5× loading buffer and 2× loading buffer?
The main difference lies in the concentration ratio and dilution ratio. For 5×, it needs to be diluted to 1× at a ratio of 1:4; for 2×, it should be diluted at a ratio of 1:1. Both have the same components and action principles, and can be selected based on experimental habits and sample volume.
