Cell Function FAQs

Agreed. Because single cells in flow assay are more sensitive and fragile, the permeabilization operation should be mild. If customers do not have saponin, they can add 1%BSA or 3-5% FBS to PBS, and then add triton X-100 with a final concentration of 0.2%, and mix it as a permeabilization reagent.

The addition of BSA can increase the osmotic pressure and ion concentration of PBS, making it more similar to the environment of intracellular fluid, thus reducing the effect of shear force on cells. In addition, BSA also has a certain buffer performance, which helps to maintain the stability of PBS, prevent the pH value of the solution from changing, and further protect the cells.

The permeability of saponin is reversible, and the saponin permeability experiment is generally used, and the subsequent washing steps will continue to add saponin to maintain cell permeability.

Theoretically, it can be done, but we have not used it. We suggest that you make sure whether this instrument can scan its absorption spectrum (230 nm~800nm) and record the data of A280 and A655 (1 cm optical path).

150,000 is the molecular weight of the antibody, and the molecular weight of different subtypes of antibodies is different. 1 Dalton =1g/mol, the unit of molar extinction coefficient is (L/mol•cm). If the customer needs to use this formula to calculate the protein concentration, they also need to know some fixed parameters of their own protein.

Fluorescence co-localization is the use of fluorescent probes and co-localization markers to link proteins to target sites, and our labeling kit allows fluorescein to bind to proteins to visualize the location of proteins. The binding site is the primary amino group of the protein for labeling, but whether fluorescence colocalization can be performed is related to the protein itself, or it needs to be determined by relevant pre-experiments.

The labeling kit can effectively label proteins containing primary amino (-NH2) molecules, but the experimental application direction of the labeled proteins cannot be determined. It is recommended that customers do pre-experiments to verify the feasibility of the experiments.

Both systems have the same resolution ratio, while one-step operation takes easier process, and two-step operation requires less reagent.

It is not suitable. The emmision fluorescence of EGFP will interfere both FITC and PE channel which are the two channels for JC-1 detection.

The method hasn't been verified.

No. The detection must be done in one hour after staining. The permeabilization and mitochondrial membrane potential will be changed by fixation.