Key operating points of CAPS buffer in protein membrane transfer

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CAPS buffer, as a very common reagent in biochemical experiments, is sometimes applied to various diagnostic kits to assist operators in testing. Recently, Desheng has found that related information on its use for protein transfer has been constantly appearing online, and many experimental personnel are not clear about its main operating points. Therefore, the following will provide a detailed introduction around this aspect.

1Preparation of CAPS in Protein Transfer Membrane Experiment

1. Generally, in experiments, 22.13g of CAPS powder is prepared based on a buffer solution of 10xCAPS (100mmol/L). Then, deionized water is added to 900ml for dissolution, followed by 2mol/L of sodium hydroxide. The pH value is adjusted to 11.0, and then the volume is fixed to 1L. It is stored at 4 degrees Celsius.

2. If CAPS imprinting buffer is required, according to 10xCAPS 200ml, the preparation method is to add 200ml of methanol and 1600ml of deionized water to obtain it.

2Key points for using CAPS in experiments

1. According to conventional conditions, CAPS is applied to proteins ≥ 20KD.

2. Ensure the concentration of methanol, which should be within the range of 0-20%. The level of methanol concentration varies with the level of molecular weight protein transfer, such as low concentration suitable for high molecular weight protein transfer.

3. After the concentration adjustment is completed, take out the PVDF film, soak it in methanol, and then place it in CAPS electrophoresis buffer. Pay attention to this operation to prevent the film from drying out, otherwise it is not conducive to the experiment.

4. Take out the electrophoresis gel and soak it in CAPS buffer for 5-10 minutes. If it is to transfer some strong basic proteins, this operation requirement can be ignored.

5. Put the filter paper and sponge into the electrophoresis buffer solution to soak in red, and then put the sponge, filter paper, PVDF film, gel, etc. into the electroimprinting interlayer for transfer printing. Please note that the transfer requirement is to transfer at room temperature under a constant voltage of 50V, with a time of about 2 hours.

6. If the extracted protein film needs to be colored, it can be lightly rinsed with deionized water, then soaked in methanol for three seconds, and then processed and dried.

In addition to taking on a significant role in protein film experiments, CAPS also holds a position in the welding materials and coating industries. As a supplier of biological buffering agents, Desheng Biochemical produces products including CAPS, TRIS, BICINE, MOPS, etc., with a purity of over 99%. The process is stable and can meet different experimental requirements. If you are interested, please click on the website to inquire about details and purchase!