Lysosome Purification by Gradient Density

Background of Lysosome Purification by Gradient Density

Most biochemical and structural analyses of lysosomes have been performed on organelles of rat liver. Accumulating evidence suggests that lysosomes are also involved in cardiac Ca²⁺ processing and play an important role in cardiac physiology. Furthermore, we currently know very little about the protein composition of lysosomes in cardiac atria. Therefore, it is imperative to develop techniques to facilitate lysosomal proteomic characterization. This can provide valuable information about the function of lysosomal pathways in normal and disease states. Density gradient-based lysosome purification is a classic method that is widely used today. Scientists have successfully isolated lysosomes from tissues and cells by differential and density gradient centrifugation, including purification of lysosomes from light mitochondrial pellets or postnuclear supernatants using Percoll, Nycodenz, and iodixanol.

Fig. 1. Flow chart of the lysosome isolation. (Ayagama T, et al., 2021)

Our Gradient Density Lysosome Purification Services

Rapid and efficient isolation of intact lysosomes facilitates further study of lysosomal functions and metabolites through proteomic analysis. For many years, CD BioSciences has developed a variety of effective strategies to isolate ultra-pure and functional lysosomes for customers worldwide. Here, we have developed flexible, low-cost, and improved density gradient methods for the isolation of lysosomes. In addition, we provide lysosomes for label-free quantitative proteomic mass spectrometry to help you study the role of lysosomes in atrial physiology and pathophysiology.

• Isolation of Lysosomes Based on Density Gradient Centrifugation

We place cell or tissue homogenates in layers on a buffer layer consisting of different concentrations of sucrose, ficoll or similar dense materials. Then centrifuged at high speed, organelles of different densities are separated into different fractions throughout the gradient, resulting in high yields of lysosomes.

In addition, we optimized this method to include purification of lysosomes by loading cells or tissues with Triton WR 1339, iron sorbitol or gold nanoparticles while accumulating in lysosomes and varying their buoyancy density.

• Isolation of Lysosomes Based on Density Gradient Electrophoresis

We can separate lysosomes, peroxisomes and mitochondria based on their different charge properties with our specialized equipment.

Why Choose Us

The density gradient electrophoresis method can achieve a level of separation that density gradient centrifugation cannot provide. Our dedicated team will consider the advantages and disadvantages of both strategies to develop the best solution for your project.

• Features of Isolation of Lysosomes Based on Density Gradient Centrifugation

• Requires a large amount of starting material to be placed.
• Provides high yields of lysosomes, but they are not completely pure and often contain microsomal or mitochondrial contaminants.
• When lysosomes are stored in diseased cells or tissues, the lysosomes redistribute or diffuse in the gradient and reduce their yield and purity.

• Features of Isolation of Lysosomes Based on Density Gradient Electrophoresis

• Needs to incorporate large amounts of starting material.
• Requires specialized equipment.
• Has the ability to purify ultrapure plasma membranes.
• Poor separation to produce enough pure material for functional lysosome assays.

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.

References

1. Ayagama T, Bose S J, Capel R A, et al. (2021) A modified density gradient proteomic-based method to analyze endolysosomal proteins in cardiac tissue[J]. Iscience. 24(9): 102949.
2. Graham J M. (2001) Isolation of Golgi membranes from tissues and cells by differential and density gradient centrifugation[J]. Current protocols in cell biology. 10(1): 3.9. 1-3.9. 24.