Chromatographic Column For Separating Exosomes (Cell Supernatant)

 

Product Name	Cat. No.	Spec.
Chromatographic Column for Separating Exosomes(Cell Supernatant)	G4113-5	5 pieces

 

 

Exosomes are nanoscale membranous vesicles with a uniform size and a particle size of 30 - 150 nm that are actively secreted into the extracellular space by cells. This vesicles have their own specific markers, and are coated with proteins, mRNA, miRNA, lipids, and other substances, which can be used as a pathway for substance and signal communication between cells and participate in processes such as cell growth, cell proliferation, cell differentiation, cell migration, angiogenesis, and tumor growth.

This product is a size exclusion chromatography column (SEC) suitable for rapid separation and purification of exosomes from cell supernatants. This product uses a cell supernatant concentration agent to separate and enrich cell supernatant exosomes, and then uses an extracellular vesicle separation chromatography column to purify the concentrated exosomes. Which can not only retain high-purity and high activity exosomes, but also efficiently remove proteins; Effectively avoiding the damage to the extracellular membrane structure caused by ultra-high centrifugal force in the ultra-high speed centrifugation method, and reduce the influence of high concentration proteins and other substances on the purity of exosomes in traditional precipitation methods.

 

 

Transportation at room temperature; Store at room temperature with a validity period of 12 months.

 

 

Component Number	Component	G4113-5
G4113-1	Chromatographic Column for Separating Exosomes(Cell Supernatant)	5 pieces
G4113-2	Cell supernatant concentrate	250mL
Product Manual		1 pc

 

 

1. Cell supernatant concentration

a) The biological samples of cell supernatant are filtered by 0.22 μM filter membrane and preliminary purified for use;

b) Mix the cell supernatant biological sample and cell supernatant concentrate in equal volume, shake vigorously for 15 seconds, and let it stand for 2 hours in a refrigerator at 4 ℃;

c) Place the mixture in a centrifuge at 4 ℃ at 10000 × g for 60 minutes; Discard the supernatant;

d) 1 × PBS buffer of 5% volume of cell supernatant biological sample is added to fully resuspension centrifuge precipitation;

e) The heavy suspension is rich in exosomes, which can be used for subsequent experiments or frozen at -80 ℃ for backup.

2. Preparation for purification of exosomes

a) Prepare a room temperature, sterile buffer solution, such as PBS (recommended G4202);

b) prepare room temperature, sterile 0.1M NaOH reagent and 20% ethanol;

c) Before using the separation column, remove the red upper cover of the separation column and place it on a bracket (recommended G6063) for use;

3. Exosomes separation

a) Prepare exosome samples after concentration and resuspension of cell supernatant concentrate for later use;

b) Remove the white bottom cover of the separation column, and after the protective liquid inside the column flows out, add a total of 30 mL of buffer solution in batches to clean the separation column;

c) After all the buffer above the sieve plate enters the separation column; Take 0.5mL of the spare exosome sample from step 3.a) and add it to the center of the sieve plate (if less than 0.5mL, add buffer solution to make up to 0.5mL);

d) After all exosome samples pass through the sieve plate and enter the separation column, add 2.5mL buffer solution;

e) After all the buffer solution in the previous step passes through the sieve plate and enters the separation column, add 1mL of buffer solution, and at the same time collect 1mL of washout solution, which is rich in naturally active exosomes; It can be applied for subsequent experiments or frozen at -80 ℃ for backup;

4. Post separation treatment

a) After completing the separation of exosomes, add a total of 30 mL of buffer solution in batches to clean the separation column;

b) If it is necessary to continue using the separation column to separate the exosomes from the cell supernatant, repeat steps 3.c) to 3.e);

c) If it is not necessary to continue using the separation column to separate exosomes from the cell supernatant, first add 5 mL of 0.1M NaOH to clean the separation column, then add a total of 20 mL of buffer solution in batches to clean and balance the separation column, and then add 5 mL of 20% ethanol to clean the separation column. After cleaning, cover the white bottom cover of the separation column; Finally, add 2mL of 20% ethanol to moisturize, cover and tighten the red top of the separation column, and store the separation column at room temperature.

 

 

1. When separating exosomes from different cell supernatants using this product, the elution curve and purity slightly differ, but it does not affect the overall trend.

2. During the process of separating cell supernatant exosomes from this product, it is necessary to avoid prolonged interruption of liquid flow in the separation column.

3. During the use of this product, do not separate biological samples from different sources on the same column, and do not reuse the same column more than 5 times.

4. This product is suitable for storage and use at room temperature, and low temperature use and high temperature heating should be avoided.

5. During the experiment, please wear lab clothes and disposable gloves to avoid contamination and ensure safety.

 

For Research Use Only!