Specifications

Product description

Exosomes are small vesicles secreted by living cells, with a diameter of approximately 30-150 nm, featuring a typical lipid bilayer structure. They exist in cell culture supernatants, serum, plasma, saliva, urine, amniotic fluid, and other biological fluids. Exosomes carry various important cargoes, including proteins, lipids, DNA, and RNA. They play a crucial role in the transfer of substances and information between cells and hold great promise as early diagnostic biomarkers for multiple diseases.

This kit employs a unique separation technology to rapidly obtain a large quantity of intact exosome particles from serum or plasma. The extracted exosomes are suitable for downstream applications such as NTA particle size analysis, Western Blot, nucleic acid extraction, and subsequent analyses (e.g., qPCR, sequencing). Note: It is not recommended for exosome labeling or protein mass spectrometry analysis.

Components

Storage

This kit should be transported at room temperature and stored for 12 months at room temperature or 2-8°C (store different components according to their specific requirements). Please mix thoroughly before use.

Instructions

Each reaction in this kit is based on 250 μL of serum or plasma as the exosome extraction sample. The scale can be proportionally increased according to the required amount of exosomes. The recommended minimum starting sample volume is 100 μL. Please refer to the table below for details:

1. Sample Pre-processing

1)For frozen samples: Thaw at room temperature or in a 25°C water bath. Place the completely thawed sample on ice.

2)For fresh samples: Collect the sample and place it on ice. Centrifuge at 3,000 × g, 4°C for 15 min to remove cells or cell debris. Transfer the supernatant to a new tube after centrifugation.

2. Exosome Precipitation

1)Pipette 250 μL (minimum 100 μL) of the processed serum or plasma into a 1.5 mL microcentrifuge tube (not provided). Add an equal volume (250 μL) of Sample Diluent and mix well. Then add 125 μL of Exosome Precipitation Reagent. Invert thoroughly to mix and incubate at 4°C for at least 30 min.

2)Centrifuge the incubated sample at 1,500 × g, 4°C for 30 min. A white pellet will be visible at the bottom of the tube. Carefully aspirate the supernatant. Centrifuge again at 1,500 × g, 4°C for 5 min to completely remove residual liquid.Note: Be careful not to disturb the exosome pellet.

3. Exosome Resuspension

Add 100 μL of Sample Diluent or the appropriate buffer required for downstream applications. Resuspend the pellet thoroughly by pipetting up and down or vortexing until the precipitate is completely dissolved. This resuspension contains the extracted intact exosomes.

4. Exosome Purification

Transfer the resuspended exosomes to a purification column placed in a 1.5 mL collection tube (provided in the kit). Centrifuge at 2,000 × g, 4°C for 5 min. Discard the purification column; the exosomes are now in the collection tube. The extracted exosomes can be used directly for downstream experiments (e.g., particle size analysis, nucleic acid extraction) or stored at 2-8°C for one week, or at -80°C for approximately three months.

Notes

1. Mix 41206-A thoroughly before use.

2. This product is specifically designed for serum/plasma samples and is not suitable for exosome extraction from other body fluids or cell culture supernatants. For exosome isolation from cell culture supernatant, please use the Cell Culture Supernatant Exosome Rapid Extraction Kit.

3. For further purification of exosomes, affinity purification using magnetic beads coated with specific antibodies is recommended.

4. For your safety and health, please wear a lab coat and disposable gloves during operation.

5. For research use only!

Documents:

Safety Data Sheet

41206_MSDS_HB260525

Manuals:

41206_Manual_HB20260127

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