1. Advantages of the Cell-Based ELISA: Cell-Based ELISA (cell-based ELISA) is a new ELISA technology with two of the most prominent advantages:
1.1 It is not necessary to extract the protein and coat the microplate: the cells are directly cultured in the microplate, and when the sample is to be detected, the cells are fixed on the microplate and permeabilized. This avoids the fact that the experimental results deviate from the actual situation to some extent due to the objective and subjective loss of the sample when extracting the protein. At the same time, the microplate is not coated, which simplifies the experimental process and helps to improve efficiency. Researchers can detect the expression of target cell proteins after stimulation or inhibition on a 96-well enzyme-linked plate. The loss of sample can be minimized by eliminating the need to extract proteins and lyse cells. This new ELISA technology can detect large amounts of intracellular proteins more quickly and conveniently than other common ELISA assays. .
1.2 Two different proteins can be detected simultaneously: after blocking, two primary antibodies against different proteins and derived from different hosts are added, then different secondary antibodies are added, and two fluorescent substrates are added to detect two wavelengths. The benefits of simultaneous detection of both proteins are obvious, reducing the amount of work and, in addition, meeting specific experimental needs. For example, to determine the phosphorylation ratio of a protein, you need to determine the amount of phosphorylated protein and total protein. Quantity, these two measurements were carried out in the same experiment, which helps to eliminate experimental errors and obtain more accurate experimental results.
2. Two technologies of Cell-Based ELISA:
Some companies have developed dual-channel Cell-Based ELISA technology;
Dual channel vs. single channel Cell-based ELISA:
The dual-channel cell-based ELISA, as the name implies, can simultaneously detect two proteins of interest at the same time. The Cell-based ELISA provided by R&D Systems is a two-channel cell-based ELISA. The principle is slightly: (requires fluorescence detection and corresponding instruments);
Single-channel cell-based ELISA, which can only detect one protein of interest at a time. The main difference between it and the normal ELISA is the sample processing process:
3. Application of cell-based ELISA:
The product, most developed to detect the relative content of phosphorylated and non-phosphorylated proteins;
4. Companies with cell-based ELISA products:
Currently, a number of elisa product providers offer cell based elisa kits such as Rnd systems, raybiotech, ebioscience, millipore, etc.;
5. How do I perform a cell based elisa experiment myself ?
In fact, according to the single-channel cell based elisa principle, you can build your own cell based elisa experimental system;
The cells are added to the culture plate;
Add stimulants or inhibitors for cultivation;
Fix or seal the cells;
Adding a first antibody;
Add HRP-conjugated secondary antibody (the choice of secondary antibody, same as western blot);
Add substrate to develop color.
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