CAS 1135-40-6 CAPS Cyclohexylamine Propanesulfonic Acid In Experiment

Cyclohexylamine propanesulfonic acid CAPS is a commonly used biochemical raw material. It can be used as a curing agent for fluxes, desiccants, and water-based isocyanates in coatings. In the biochemical industry, it is usually used as a biological buffer. According to its purity and impurity content, the application industry is also different.

From the point of view of molecular structure, CAPS is a cyclohexane N-substituted sulfamic acid with a molecular weight of 221.32 and amphoteric. It is a very good biological buffer with a pH buffer range of 9.7-11.1. It is usually used for WB experiments. Buffer for protein PVDF transfer membrane or nitrocellulose NC transfer membrane.

WB stands for western blot protein immunoblotting experiment. The principle is to color cells or biological samples treated by gel electrophoresis with specific antibodies, and obtain the expression of specific proteins in the analyzed cells or tissues by analyzing the position and depth of the coloring. Information. The experimental notes are as follows:

CAPS reagent

1. SDS-PAGE electrophoresis: similar to the nucleic acid hybridization method, except that the detected is the protein, the probe is the antibody, and the colored secondary antibody is labeled. Generally speaking, Tri-Glycerine or Tris-Tricine buffer system is used for low molecular weight proteins, and CAPS buffer system is better for high molecular weight proteins. Glycine in the buffer will affect sequencing, so the transfer buffer used for sequencing is 10mM/1CAPS10% methanol. The concentration of methanol in the CAPS electroimprinting buffer is 0-20%. Generally, a buffer with a high methanol concentration has a good transfer effect on low-molecular-weight proteins, while a buffer with a low methanol concentration or even no methanol is good for high-molecular-weight proteins. Transfer.

2. When PVDF membrane is processed, soak it in methanol for a few seconds, and then put it in CAPS electroimprinting buffer. Follow-up operations must prevent the membrane from drying out. If the PVDF membrane becomes dry, repeat the previous steps. The electrophoresis gel also needs to be soaked in CAPS buffer for 5-10 minutes. Some strongly basic proteins do not need this step.

3. The transfer conditions are usually 50V constant voltage, 100-170mA current intensity, and electroimprint transfer at room temperature. The bubbles between the gel and PVDF membrane must be removed. For proteins with higher molecular weight, pay attention to prolong the transfer time.

4. Before dyeing, PVDF membrane needs to be taken out and rinsed with deionized water, soaked in methanol, then dyed with 0.1% Coomassie brilliant blue for 30-50 seconds, decolorized with 50% methanol, washed with deionized water and dried.

The above introduces some matters needing attention in the protein imprinting experiment. In addition to the experiment itself, Desheng Technology reminds you that for safety and health, please wear laboratory clothes and wear disposable gloves for operation. The CAPS electro-imprinting buffer is recommended to be produced by Desheng The configuration of high-quality cyclohexylamine propanesulfonic acid.