Background of Lysosomal Fractions Purity Determination
Lysosomes are mainly responsible for degrading unnecessary substances in cells and can also serve as signaling organelles. Current prevailing biomedical research does not provide spatial information on lysosomes and therefore how lysosomes regulate their cellular functions. Therefore, measuring lysosome composition and its ability to change in response to stimuli will provide important information for understanding its function. Traditionally, lysosomes have been isolated by differential centrifugation or multi-step density gradient ultracentrifugation. However, these methods have some inherent disadvantages, such as impure separation, often low concentrations making some downstream analyses challenging, and time-consuming resulting in changes in the composition of organelles. The separation of lysosomes by superparamagnetic nanoparticles also has some unexpected effects on lysosome function. Therefore, determination of the purity of lysosomal fractions can ensure the accuracy of the results.
Fig. 1. Western blot analysis of fractions obtained throughout the lysosome isolation. (Ramirez-Montealegre D, et al., 2005)
Our Lysosomal Fractions Purity Determination Services
Rapid isolation of high-purity lysosomes facilitates the analysis of metabolic activity and metabolites within the lysosome lumen. For many years, CD BioSciences has focused on developing relatively simple and rapid methods to analyze the purity of lysosomal fractions. Here, we provide the following strategies to determine the purity of lysosomal fractions:
- Determination of the purity of lysosomal fractions based on Western Blot
We used standard western blotting protocols (10% gels) to analyze the purity of lysosomal fractions. This anti-lysosomal enzyme antibody does not necessarily only label lysosomes, but may also recognize immature precursor cells of the enzyme. In addition, we reflect lysosomal protein content by measuring lysosomal enzyme activity in vitro. - Determination of the purity of lysosomal fractions based on enzyme assays
We used fluorescent or spectrophotometric substrates for lysosomal enzymes to confirm purity, which can be performed immediately after purification. - Determination of the purity of lysosomal fractions based on electron microscopy
We illustrated the purity of lysosomal fractions by means of images. The centrifugation speed is mainly low enough to pellet the lysosomes and elute the lysosomes on the column directly into the electron microscope fixative solution.
Why Choose Us
- High purity and high quality lysosomes can be obtained as starting material.
- Contributes to lysosomal proteome analysis.
- Western blotting material is usually frozen within a few days to confirm purification.
- The enzyme assays method can be performed immediately after purification and requires only very small amounts of starting material.
- These methods allow rapid determination of purity.
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
- Ramirez-Montealegre D, Pearce D A. (2005) Defective lysosomal arginine transport in juvenile Batten disease[J]. Human molecular genetics. 14(23): 3759-3773.