Cell-Based Colorimetric ELISA Kits
The Cell-Based Colorimetric ELISA Kit allows for the determination of different levels of target proteins under certain experimental conditions or the effect of various stimulation methods on target protein expression in different cell types. Traditional Western blot analysis is time consuming and only semi-quantitative, while the Cell-Based Colorimetric Protein ELISA is a fast and convenient method to determine relative protein levels among various cell types. Cell-based ELISAs are performed in 96-well microplates, are scalable, and conserve cell culture and treatment reagents. The relative amount of target protein is determined using target-specific primary antibodies and HRP-conjugated secondary antibodies as the detection agent. Following the colorimetric measurement of HRP activity via substrate addition, the crystal violet whole-cell staining method is used to determine cell density. After staining, the results can be analyzed by normalizing the absorbance values to cell amounts, by which the plating difference can be adjusted. In addition to quantifying normal cellular protein concentrations, the Cell-Based Colorimetric ELISA can also be applied to detect proteins that undergo modifications including but not limited to phosphorylation, acetylation, and methylation.
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Cell-Based Fluorogenic ELISA Kits
The Cell-Based Fluorogenic ELISA Kit allows for the determination of different levels of target proteins under certain experimental conditions or the effect of various stimulation methods on target protein expression in different cell types. Traditional Western blot analysis is time consuming and only semi-quantitative, while the Cell-Based Fluorogenic Protein ELISA is a fast and convenient method to determine relative protein levels among various cell types. Cells are first seeded onto 96-well plates and then subject to various types of experimental conditions dependent upon the purpose of the experiment. The relative amount of target protein is determined by using target-specific primary antibodies and Dylight-647-conjugated secondary antibodies as detection agents. Anti-GAPDH antibodies are used for the detection of GAPDH expression level as an internal control, with FITC-conjugated anti-mouse secondary antibodies used for detection. The fluorescent intensity of FITC and DyLight-647 can be measured by fluorescent microplate readers. Ultimately, the fluorescence intensity is proportional to the concentration and amount of the proteins inside the cells. The ratio of DyLight-647 intensity versus FITC intensity from before and after treatment is able to reflect the expression changes of target proteins. Therefore, the Cell-Based Fluorogenic ELISA Kit provides a fast, easy and convenient way for high throughput screening. While the Cell-Based Fluorogenicc ELISA is effective in quantifying normal cellular protein concentrations, its most advanced application is emphasized in the HT-screening of proteins that undergo various types of modifications such as phosphorylation and acetylation.
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