Background of Enzymatic Readouts of Phagosomal Function
Phagocytes phagocytose and destroy harmful microorganisms in the newly formed phagosome and play an important role in the innate and acquired immune response. The role of phagosomes is to transport particulate material into the hydrolytic environment, which leads to its degradation. Massive protein hydrolysis is a useful indicator of the maturation process of phagosomes, as it depends on effective acidification and delivery of lysosomal enzymes to mature phagosomes for optimal activity. Thus, lysosomal hydrolase delivery and activation are important properties associated with the phagosomal maturation process. Current assays of phagosome maturation remain insensitive and few in number. The researchers hoped to assess phagosome maturation by measuring the extent of bulk proteolytic activity in the phagosome in real time.
Fig. 1. The cellular compartment of phagocytosis. (Nüsse O, 2011)
Enzymatic Readouts Services for Phagosome Function
Methods to analyze phagosome maturation as a dynamic process are rather limited. For many years, CD BioSciences has been working on the determination of important parameters of lysosomal biological function to assess the degree of phagosomal maturation, including phagosome and lysosome content, histone L activity, and lipolytic mixing. These parameters reflect the kinetic characteristics of phagosomes at different time periods and can further quantify the important biological properties of phagosomes at maturation.
Our bioscientists develop customized procedures for a range of enzyme activity assays to probe the biology of phagosomes after innate or acquired immune stimulation activates cells, including bulk protease and cysteine protease activity, lipolysis and β-galactosidase activity. The enzymatic assays available here for phagosomal function include, but are not limited to:
- Measurement of the Proteolytic Capacity of Isolated Lysosomes
- Real-Time Measurement of Proteolytic Activity in Phagolysosomes
- Real-Time Measurement of Lipase Activity in Phagosomes
- Real-Time Measurement β-galactosidase Activity in Phagosomes
- Accumulation of Pro-Cathepsin D in Phagosomes
Why Choose Us
- Can be performed within 1-4 hours.
- The sum of multiple steps in the phagosome maturation process can be reported.
- Support of multiple advanced instruments, spectrofluorometer, fluorescence plate reader and flow cytometer.
- Suitable for quantitative analysis in real-time live cell confocal microscopy.
- With a high degree of temporal detail. Ability to quantitatively characterize the onset and extent of proteolytic activity and its changes in individual phagosomes.
CD BioSciences can meet any reasonable needs of our clients, taking time and budget into consideration for you. Our aim is to be customer-centric and to provide the highest quality services to customers. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.
Reference
- Nüsse O. (2011) Biochemistry of the phagosome: the challenge to study a transient organelle. ScientificWorldJournal. 11:2364-81.