Efficient protein-to-membrane transferring buffer solution and preparation method thereof

Abstract

The invention discloses a high-efficiency protein transfer membrane buffer, which comprises the following components: the concentration of Tris is 10mM-100mM; the concentration of taurine is 10mM-200mM. Compared with the traditional membrane transfer buffer commonly used at present, the high-efficiency membrane transfer buffer of the present invention uses a taurine component with a stronger buffer capacity, which improves the buffer capacity of the buffer during electrophoresis, and is suitable for proteins with a wide range of molecular weights. Molecules have high membrane transfer efficiency, which can shorten the time for protein transfer to the membrane. At the same time, the heat generated by the buffer during electrophoresis is low, and there is no need to use ice packs to cool down, which simplifies the operation during membrane transfer. The concentration of methanol is lower, and a better transfer effect can be achieved at a concentration of 10%.

Description

technical field [0001] The invention relates to a high-efficiency protein transmembrane buffer and a preparation method thereof. Background technique [0002] Western blotting, Western Blot, is a common experimental method in molecular biology, immunogenetics and biochemistry. It is to transfer the electrophoresis-separated cells or tissue proteins from the gel to a solid support, such as nitrocellulose. A protein detection technology that uses plain film (NC film), polyvinylidene fluoride film (PVDF film), and specific antibodies to detect specific antigens. Now it has been widely used in antibody activity detection, early disease diagnosis and protein level expression research and other aspects. Membrane transfer is an important step in Western blotting experiments. Proteins separated by polyacrylamide gel (PAGE) electrophoresis need to go through the membrane transfer step, that is, transfer from the PAGE gel to the solid phase support, before they can be processed by va...

AI Technical Summary

Problems solved by technology

When using this transfer buffer system to transfer proteins, the temperature of the electrophoresis solution will increase significantly with time, high temperature will reduce the transfer effect, and will also cause displacement of PAGE gel and solid support, deformation of protein bands, etc. , so it is often necessary to add an ice pack or transfer the membrane on ice to reduce the high temperature generated during the transfer

In addition, the Tris-Glycine transfer buffer has a low effect on the transfer of high molecular weight proteins, and can only transfer part of the large molecular weight proteins from the PAGE gel to the solid support.

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