Exosome Protein Extraction Kit
Vorteile im Überblick
- High Protein Yield
- Effective Lysis
- Easy to Use
- Accurate Quantification
- Western Blot Ready
Produktnummer:
S5328.0005
Lieferzeit: 3 - 8 Werktage
Für genaue Informationen zum Liefertermin wenden Sie sich bitte an Genaxxon.
Shipment: not cooled. Store at +15°C to +30°C. For laboratory usage only!
Lieferzeit: 3 - 8 Werktage
Für genaue Informationen zum Liefertermin wenden Sie sich bitte an Genaxxon.
Shipment: not cooled. Store at +15°C to +30°C. For laboratory usage only!
Produktinformationen "Exosome Protein Extraction Kit"
Genaxxon's Exosome Protein Extraction Kit is especially engineered to extract proteins from various exosomes with high efficiency. The kit is the perfect solution for researchers aiming for high-quality Western Blot results.
Key Features:
- High Protein Yield: Achieve a high concentration of exosome proteins, ensuring optimal results for your Western Blot experiments.
- Effective Lysis: The kit contains a special protein denaturant that effectively lyses exosomes, maximizing protein extraction.
- Easy to Use: Simply mix the kit with your exosome solution to lyse the exosomes. The lysate can then be directly loaded into SDS-PAGE for straightforward analysis.
- Accurate Quantification: Measure the concentration of exosome proteins in the lysates using either the Bradford or BCA protein concentration assay methods.
- Western Blot Ready: The denatured proteins are perfectly suited for Western Blot applications, ensuring reliable and high-quality results.
Applications:
- Western Blot Analysis: Ideal for researchers focused on obtaining high-quality Western Blot data.
- Protein Quantification: Compatible with Bradford and BCA assay methods for accurate protein concentration measurement.
FAQs
Keine Suchergebnisse vorhanden.
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.
Exosomes are quite fragile and can be easily damaged or altered by harsh mechanical or chemical processes found in many commercial exosome isolation kits. This can negatively impact downstream applications such as Next-Generation Sequencing (NGS).
Here are steps you can take to minimize damage and maintain the integrity of purified exosomes:
- 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.
To improve the overall purity of exosomes a Multi-Step Purification (combining different purification methods) method can be applied. E.g. using ultracentrifugation followed by density gradient
centrifugation and size exclusion chromatography can effectively remove various types of contaminants.
Contamination from microorganisms can compromise the quality of your samples. To prevent and address this issue, follow these guidelines:
By adhering to these practices, you can minimize the risk of microbial contamination and maintain the quality of your samples.
- 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.
Exosomes are known as cell-secreted natural nanovesicles, and the low-yield of exosome production from MSC-conditioned medium under conventional culture conditions limits their applications. The optimal cell culture medium can enhance exosome secretion from MSCs.
For exosome secretion from adipose-derived stem cells, 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 our ADSC Exosome HY medium (C4161) to remove residues of the original medium. Then the ADSC cells are cultured in our exosome enrichment medium ADSC Exosome HY medium for 3-4 days before the conditioned medium can be used for exosome isolation.
For exosome purification, we recommend starting by preparing a conditioned medium after cell culture (e.g. using our ADSC Exosome HY Medium. The supernatant is then collected from cultured cells, and large fragments of cell debris, waste products produced by cell metabolism, as well as impurities in the culture medium are removed through any exosome isolation method (e.g. using our exosome isolation kits).
In general, commonly used supplements like serum or human platelet lysate contain exosomes. Therefore, we do not recommend using supplements in the preparation of conditioned medium for exosome isolation.
When producing exosomes using our ADSC eXOrich medium, there is no need to add common nutritional supplements like serum or human platelet lysate. However, users can decide to add appropriate nutritional supplements based on their specific needs.
special protein denaturant that effectively lyses exosomes
Applikation / Application:
Western Blot, Protein Quantification
Einheiten / Units:
Quelle / Source:
Sicherheits Hinweise / Safety
Klassifizierungen / Classification
eclass-Nr: 32-16-04-05
Dokumente - Protokolle - Downloads :
Hier finden Sie Informationen und weiterführende Literatur. Für weitere Dokumente (Zertifikate mit weiteren Lotnummern, Sicherheitsdatenblätter in anderer Sprache, weitere Produktinformationen) wenden Sie sich bitte an Genaxxon biosience unter: info@genaxxon.com oder Tel.: +49 731 3608 123.
Dokumente:
Produktbeschreibung
Hier finden Sie Artikel und Literaturzitate, in denen die Autoren auf die hohe Qualität dieses Genaxxonprodukts vertrauen.
Quelle: NCBI PubMed
Quelle: NCBI PubMed
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