ADSC Exosome HY Medium
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Product information "ADSC Exosome HY Medium"
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Product Details:
Benefits:
Application:
For exosome production, we recommend culturing ADSCs in dishes or flasks under regular culture conditions up to 90% confluence. Then remove the original culture medium and wash the cells with PBS and ADSC Exosome HY medium to remove residues of the original medium. Then the ADSC cells are cultured in ADSC Exosome HY medium for 3-4 days before the conditioned medium can be used for exosome isolation.
Picture 1: Flow Cytometry - Verification of CD9, CD63, CD81 surface marker
Picture 2: Yield comparison between standard cell culture media and ADSC Exosome HY Medium
Produced by Genaxxon bioscience GmbH, founded in 2002 by Dr. Norbert Tröndle to provide reliable products for PCR and custom PCR and cell culture media formulations, ADSC Exosome HY Medium offers a dependable and efficient solution for your cell culture and exosome research needs. Optimize your experiments and achieve consistent, high-quality results with a medium designed specifically for your requirements.
Conventional Methods for Exosome Extraction and Isolation
Workflow with Magnetic Beads for Exosome Purification
Low Yield of exosomes can have different reasons of which some are listed here:
Exosome amount or concentration: Use samples contain more exosomes, e.g. concentrate samples or increase sample volume if possible.
Storage: Exosomes are very susceptible to repeated freeze-thaw cycles or to high storage temperatues. Always keep exosomes at -80°C for long term storage and do not allow repeated freeze-thaw cycles.
Kit related issues: Always centrifuge your sample at 16000xg before filtering it to get rid of the cell debris and to avoid clogging the filter membrane in the next step. Filter the centrifuged sample through a filter with a pore size between 0.8µm and 0.2µm.
If you use magnetic beads for further purification, the proportion of reagent volume is crucial for optimal efficiency. For this do not use less beads (less bead volume) than recommended and always use the recommended amount of reagent for incubation and washing. Incubation mode and elution intensity should be carried out in strict accordance with the instructions.
Sometimes the incubation time between samples and magnetic beads is too short. In such a case a prolonged incubation time can increase the exosome amount isolated as can an increased vortex intensity and vortex time.
- Use Gentle Isolation Conditions: Employ low temperatures and mild centrifugal forces during the isolation process. Avoid high-speed centrifugation steps that could cause exosome rupture.
- Optimize Buffers: Use buffers that protect the structural integrity of exosomes, such as phosphate-buffered saline (PBS) or Tris-buffered saline (TBS). Ensure the pH and osmolarity of the buffers are suitable for exosome stability.
- Proper Storage Conditions: Store exosomes at low temperatures, preferably -80°C, to prevent degradation. Avoid repeated freeze-thaw cycles as they can damage the exosomal membrane.
- Quality Control Assessments: Regularly perform quality control assessments such as transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blotting. These assessments help check the morphology, size distribution, and protein markers of the exosomes to ensure their integrity.
Different isolation methods have varying capabilities to remove contaminants. For instance, immunocapture can use specific antibodies to bind exosomes, thereby reducing contamination from other components. Size exclusion chromatography (SEC) can also effectively separate exosomes from smaller particles.
- Strict Protocol Adherence: Maintain a clean work environment and use sterile techniques to avoid cross-contamination during the process. Use disposable centrifuge tubes and filters, and ensure all equipment is cleaned and calibrated regularly.
- Using the Right Filter Membrane: Ensure that the final filtration step is carried out in a clean bench. Use filter membranes with pore sizes no larger than 0.22 µm to effectively remove microorganisms.
- Using Specialized Kits and Reagents: Utilize specialized kits and reagents that are designed to address contamination issues. These products simplify the process and significantly reduce the chances of contamination.
with 4.0 g/L D-Glucose
with 4.0 mM L-Glutamine
without Phenol Red