product description
Extracellular vesicle concentrate is an efficient concentration reagent suitable for large volume extracellular vesicle samples, which can quickly process large volume samples. The concentrated sample can be used for subsequent experiments such as exosome isolation and purification, RNA extraction, protein extraction, etc.
This product is mainly used for the concentration of extracellular vesicles in urine and cell culture supernatants, but the concentrated extracellular vesicles are not suitable for direct NTA particle size analysis, Western Blot, qPCR, NGS sequencing, extracellular vesicle labeling, or protein mass spectrometry analysis.
ordering information
product name | Item number | specifications |
Extracellular vesicle concentrate | AC25L141 | 50mL |
Extracellular vesicle concentrate | AC25L142 | 100mL |
Transportation and storage
Transport at room temperature. Stored at 4 ℃, with a shelf life of 12 months.
Usage
Self provided instruments, reagents, and consumables: high-speed freeze centrifuge, 50mL centrifuge tube, sterile PBS.
1. Sample pretreatment
(1) Cell removal: For samples containing cells, centrifuge at 4 ℃, 300g for 5 minutes, and transfer the supernatant to a new centrifuge tube; Cell free samples can skip this step.
(2) Remove cells and cell debris: Centrifuge at 4 ℃, 2000g, for 10 minutes, and transfer the supernatant to a new centrifuge tube.
(3) Remove large volume particles: Centrifuge the supernatant obtained in step (2) at 4 ℃, 14000g, for 30 minutes, and transfer the supernatant to a new centrifuge tube. Alternatively, choose an alternative method: filter the supernatant from step (2) through a 0.2 μ m microporous membrane and collect the filtrate.
2. Concentration of large volume extracellular vesicle samples
(1) Add 24mL of preprocessed sample into a 50mL centrifuge tube, add 6mL of extracellular vesicle concentrate, invert and mix thoroughly. Note: The ratio of sample volume to exosome concentrate is 4:1, and the sample volume can be adjusted according to this ratio. To ensure effectiveness, it is recommended that the concentrated sample volume be no less than 24mL.
(2) Incubate at room temperature for 60 minutes (rapid process); If time permits, it is recommended to incubate overnight at 4 ℃ (high recovery process).
(3) Centrifuge at 4 ℃ and 12000g for 30 minutes, discard the supernatant. After carefully discarding or pouring out the supernatant, use a 200 μ L pipette to aspirate as much residual supernatant as possible.
3. Resurrected extracellular vesicle samples
(1) Add an appropriate amount of sterile PBS to the precipitate, gently blow it up and down with a pipette, and mix thoroughly. Note: The volume of sterile PBS can be flexibly adjusted, but the concentration factor is not recommended to exceed 50 times. It is recommended to concentrate 24-30 times. If this kit is used for extracellular vesicle separation, add 10mL of cell culture supernatant/urine sample to 100 μ L of sterile PBS for resuspension.
(2) After the extracellular vesicle sample is completely dissolved, it can be used for subsequent experiments such as extracellular vesicle isolation and purification, RNA extraction, protein extraction, etc. Note: If subsequent experiments are not carried out immediately, the dissolved samples can be stored at -80 ℃ (if experiments are conducted within 24 hours, they can be stored at 4 ℃). For extracellular vesicle separation and purification, precipitation method extracellular vesicle separation product (item number: AC15L412), SEC method extracellular vesicle separation product (item number: AC25L434), or magnetic bead method extracellular vesicle separation product (item number: AC25L422) or (item number: AC25L423) can be selected.
precautions
1. This product is only for scientific experimental research and should not be used in clinical diagnosis, treatment, or other fields.
2. For your safety and health, please wear lab clothes and disposable gloves when operating.
3. During use, try to open the bottle cap in a sterile environment to reduce contamination of the reagents.
Related product recommendations
Quick Cell Exosome Extraction Kit (for Cell Supernatant) (Product Code: AC15L412)
Extracellular vesicle isolation kit (SEC method) (item number: AC25L434)
Extracellular vesicle isolation kit (magnetic bead method) (item number: AC25L422)
Extracellular vesicle tracer kit (item number: AC25L166)
S-Buffer Replacement Kit (Product Code: AC25L491)
M-Buffer Replacement Kit (Product Code: AC25L492)
This product is only for scientific experimental research and should not be used in clinical diagnosis, treatment, or other fields.
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