Phosphodiesterase, Phosphatase, Kinase, and ELISA Assay Development
Assays and Applications
Laboratory assays are necessary for determining the mechanisms of action of drugs and compounds used on cells. Whereas IC-50 and cytotoxicity assays are focused on the viability of cells, other kinds of assays give information regarding protein expression and other internal properties that are not directly tied with cytotoxicity. These assays are important for research studies regarding specific cellular pathways, and are a widespread in clinical fields.
Phosphodiesterase, phosphatase, kinase, and ELISA assays are some of the more common assays used for determining specific cellular processes. Phosphodiesterase assays are used to determine the presence of phosphodiesterases that can break apart the phosphate backbone of DNA. They can be used in oncology studies evaluating the degradation of genomic DNA and potential defects that can occur in cells. Phosphatase and kinase assays are used to determine the phosphorylation activities inside of cells; whereas phosphatase removes phosphate groups, kinase catalyzes their addition. Such assays are primarily used in research pertaining to post-translational modifications in cells, and can provide insight into epigenetic factors that affect cellular activities. ELISA assays are a broad category of assays that are used to determine the presence of specific proteins; ELISA stands for Enzyme-Linked Immuno-Sorbent Assay, and it refers to the usage of linked antibodies for the signaling of protein presence. Specially selected antibodies are used to first adhere to proteins, and then they are tagged with additional antibodies that contain fluorescent markers that can be used for detection procedures.
The assays mentioned above are crucial to biological studies involving protein expression and the effects of drugs on cells. Although a drug may show beneficial effects in vivo, it is important to understand the pathway used by the drug and what other potential effects may result. This is where assays come in, as they can provide quantifiable information regarding the specific activities of given drugs and the effects on protein expression, epigenetic factors, and overall cellular function.
Altogen Labs Assay Services
Altogen Labs offers kinase, phosphatase, and phosphodiesterase assay development services for screening and validation studies. For each project, an individual pathway is defined that matches the biology and chemistry requirements of the target to the most appropriate assay technology platform available. Company scientists have experience developing assays with all major fluorometric readouts and biologically relevant formats. Specific assays can include fluorescence, time-resolved fluorescence resonance energy transfer, luminescence, absorbance, calcium mobilization (FLIPR), ELISA, cellular imaging, fluorescent microscopy, enzyme assays, reporter assays, viability assays, and protein translocation. Compound profiling is supported by qRT-PCR, Western blot, FACS and other expression and functional assays.
Laboratories seeking to gain quantifiable and reproducible results should consider outsourcing detection procedures to companies with clearly-defined and pre-tested protocols. Formalized assays from GLP-compliant laboratories are trustworthy for research papers, and can save time for researchers. Many times, cells can be modified in laboratories, and then the cells can be sent off for testing – resulting in guaranteed results that are easy to use in data analysis.
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