Using a FLIPR Assay
To perform FLIPR assays, the cell density must be optimized and stable. One way to assess the cell density is to seed the whole plate at one density. Then, perform two additions of cells at a maximum against concentration and buffer. Once this has been done, a single-point screen should be performed.
Then, a second step is to examine the cell kinetics. To assess the stability of the signal, the optimal dye concentration, agonist, and cell density must be determined. The flipr assay is used to perform a variety of roles and in this article we will look at these roles more closely.
Understanding More About The Process Involved
CHO cells are commonly used as test cells. Other cell lines, such as AV12 and HEK293 cells, can also be used. Some of these cells can benefit from poly-D lysine-coated plates. Other settings can be optimized to improve the FLIPR(tm) variable signal. For best results, the sample volume should be at least 1.6 mL. For the best result, use a 96-well plate with the same size and spacing.
A 96-well FLIPR assay uses a three-well plate. Choosing the best cell culture medium is crucial for the success of FLIPR assays. CHO cells require 37degC to aspirate the sample. HEK293 cells should be dyed at 25degC. Alternatively, AV12 cells should be dyed at 25deg C. Ensure that the poly-D lysine-coated plates have no effect on the results.
During the FLIPR assay, cells must be cultured in a confluent monolayer. The optimal seeding density depends on the cell type and time in culture. A cell-seeding density that is optimized for this procedure is important for monitoring the functional activity of the cells. This means optimizing the seeding density to avoid the inhibition of cell growth. It is critical to optimize the number of samples that are used during an experiment.
Requirements For Drugs And Medicine
A FLIPR assay requires cells to be cultured in a monolayer. The number of cells needed depends on the concentration of compound A and the drug-selection condition. A confluent cell monolayer should have a uniform color in order to measure the response to drug. Depending on the density of the cell monolayer, a thin layer may be formed. Then, the dye or liquid should be removed manually.
After the FLIPR assay is complete, cells are cultured in growth medium with 20 ul of Ca3 dye. Then, beta-alanine at 10 M final concentration is added to the medium. The cells are cultured for a total of 30 minutes. These assays have several advantages. In addition to being more effective than other types of FLIPR assays, they are easier to use and perform.
The FLIPR assay is useful for screening various compounds in the cell. It is very flexible, and it allows researchers to detect all three types of compounds simultaneously. It is easy to perform and is compatible with all types of matrices. A FLIPR assay has an advantage over many other methods. The sensitivity of this assay is greater than with other assays. The results obtained with FLIPR assays are more accurate.
