GTOnco™ Apoptosis Assay
For cancer therapy, apoptotic tumor cells are able to prime the anti-tumor immune response and increase the proliferative ability of tumor-specific T cells. In addition, the immune system to some degree relies on apoptosis to prevent autoimmunity and immunopathology. At the end of the immune response, apoptosis drastically reduces the numbers of activated T cells. Apoptosis is a highly regulated process that affects the anti-tumor immune response of gene therapy-based I-O products. At Creative Biolabs, we have developed a large number of apoptosis assays devoted to the identification of apoptotic cells and the analysis of the biochemical, morphological, and molecular changes during this biological process.
Figure 1. Schematic representation of apoptotic events. (Elmore, 2007)
Apoptosis is considered as a vital component of various processes including proper development and functioning of the immune system, and normal cell turnover. Creative Biolabs provides a wide array of apoptosis assays for our clients across the world to measure multiple components changes in their gene therapy-based I-O products development. At GTOnco™, our featured apoptosis assays include but not limited to:
- Preliminary Assays for Cell Death Detection
The initial indication for the occurrence of cell death is detected by MTS, Vital Dye Exclusion, or Hoechst staining. Meanwhile, these initial observations will be followed-up by more specific assays.
- Plasma Membrane Changes Detected by Annexin V Binding Assays and Staining
Generally, by the reaction of phosphatidylserine with Annexin V-fluorochrome conjugates on the plasma membrane, the changes in plasma membrane composition and function are detected. This method can distinguish between the early and late apoptotic events with the combination of propidium iodide (PI) staining.
- Morphology and DNA Fragmentation Detected by Flow Cytometry-Based Assays
We provide quick and reliable detection methods to recognize the apoptotic cells, which are based on their reduced DNA-associated fluorescence in cells with diminished DNA content (sub-G1) or morphological changes. In addition to the flow cytometry-based assays, trypan blue, or Hoechst staining are also available at GTOnco™.
- Caspase Activity Determination
The best-recognized biochemical hallmark of early stage apoptosis is the activation of caspase enzymes. The activated caspases cleave many cellular proteins and the resulting fragments may serve as apoptosis markers recognized by a variety of assays, such as colorimetric/fluorometric substrate-based assays, flow cytometry and western blot analysis.
- Other Assays Available
Such as mitochondrial membrane potential assays, nuclear condensation, and cleavage of anti-apoptotic Bcl-2 family proteins are available at GTOnco™.
At Creative Biolabs, we can provide superior performance-highly sensitive assays for our clients to detect the different phase of apoptosis. Contact us today for a quotation or any question. Our customer service representatives are available 24hr a day!
Reference
- Elmore, S. (2007). Apoptosis: A Review of Programmed Cell Death. Toxicologic Pathology. 35(4), pp.495-516.
