Product Details

Description

      Applicable to the initial milk such as cow milk, goat milk, horse milk and camel milk with a volume of 400 - 500 mL.

      Exosomes are small vesicles(30-150nm) containing RNA and protein secreted by cells, which are abundant in body fluids such    

     as blood, saliva, urine and milk. Exosome is considered to have the function of the inter-cellular messenger, transmitting their       

     effectors   or signal molecules between specific cells. However, its structure, composition of effectors and the biological   

     pathway involved currently remain unclear.

     In the biological functional study of exosome, it is necessary to separate its complete particles, but the conventional   

     ultracentrifugation method has complicated steps, high requirements to the hardware and character of difficult operation. The   

     Rapid Extraction Kit of Exosome independently developed by Umibiois suitable for the extraction of exosome in the emulsion  

     after  the optimization of the components, which can obtain high-purity exosomal particles rapidly and efficiently. It can be applied 

     to electron microscopy analysis, NTA particle size analysis, nucleic acid analysis, protein analysis, cytological experiments, 

     animal experiments, etc.

Self-Provided Materials 

  High-speed centrifuge (up to 10000 g centrifugal force), vortex oscillator; 50 mL centrifugal rotor, 50mL centrifugal tube, 

  2 mL centrifugal rotor, 1.5 mL centrifugal tube, PBS buffer solution. 

Product Components

    Component Name                                          Spec

    Solution A*                                                     50 mL

    Solution B*                                                     60 mL

    Solution C*                                                     60 mL

    Solution D*                                                    120 mL

    50 mL Centrifugal Filter Column                    20pcs

     * Nuclease-free, Sterile

Advantages of the Product Series

• Covers cell supernatant, tissue, urine, serum, plasma, milk, etc.

• High sample processing capacity, up to 5 times that of similar products.

• Exosomes can be used for cell co-culture and omics research.

• Dedicated to the continuous optimization of exosome kits for research and development.

• Trusted by over 300 customers, ensuring worry-free purchasing and use.

• Cost-effective to support large-scale research while saving costs.

Operation Procedures 

       I. Sample Pretreatment

      1. Sampling: in case of the frozen sample, take it out from the fridge and thaw it in a 25 °C water bath, and place the 

       fully-thawed sample on the ice; in case of the fresh sample, collect the sample and place it on the ice.

      2. Initial dosage of the sample: (sample dosage for a single extraction)

                    Sample Name                              Optimal Treatment Volume for a Single Time

                       Emulsion                                                                  50 mL

      3. Centrifugation to remove the lipids: transfer the sample to the centrifugal tube and centrifuge at 4℃ at 10000 g for 

      20 min to remove the lipids and some proteins from the sample;(Note: the sample is divided into three layers after     

      centrifugation, with the upper layer being a lipid layer, the lower layer being a protein precipitate, and the middle layer   

      being whey). After centrifugation, the state of the upper layer is “compact, stable and not easy to fall off”. If the upper 

      layer is “fluffy and easy to fall off” and there is a lot of precipitates in the lower layer, this step can be repeated 

      continuously and the middle layer liquid shall be taken after each centrifugation)

      4. Transferring the whey: transfer the whey (middle layer liquid) whose lipids have been removed to a new 50mL centrifugal   

      tube;  (Note: the upper layer of lipids can be punctured by a spearhead and slowly dumped, or transferred with a 

      pipette. It is a normal phenomenon that there is a small amount of lipids and precipitates in the transferred whey, 

      which will not affect subsequent experiment.)

      II Impure Protein Removal

      1. Settling the whey: add Solution A into the whey. Invert the centrifugal tube to mix it thoroughly until it is “translucent”. Then     

      add Solution B and invert the tube to mix thoroughly. Let it stand at 2℃to8℃ for 10 min; (Note: after static placement is    

      completed, shake gently the centrifugal tube. “Tofu pudding-like” solid will be presented and the fluid will be  

      “transparent”. If the state of “tofu pudding” is not presented or the sample is still “milk white”, appropriate amount of   

       Solution B can be added until the fluid turns “transparent”.)

                           Whey Volume               Solution A Dosage               Solution B Dosage

                                40 mL                               4 mL                                        3 mL

      Note: please convert specific dosages to be added according to the proportion of the table above.

      2. Centrifugation to remove the protein: centrifuge the settled whey at 4℃ at 10000 g for 10 min and collect the supernatant;

      3. Filtering the supernatant: transfer the collected supernatant to 50 mL centrifugation and filtration column and centrifuge at 4℃  

      for 2 min at 3000 g; (Note: if the supernatant is not fully filtered, this step can be repeated. 50mL centrifugation and    

      filtration column is a disposable material and repeated use is not recommended.)

      4. Transfer the filtered supernatant to the new centrifugal tube, add Solution C and invert it to thoroughly mix it; (Note: the    

      dosage of Solution C shall be consistent with the dosage of Solution B)

                          Sample Volume after Centrifugation and Filtration                                     Solution C Dosage

                                                             40 mL                                                                                        3 mL

     III Exosome Extraction

     1. Pretreating the supernatant: add Solution D into the supernatant added Solution C, and the specific dosages are as follows:

                         Sample Dosage                                                             Solution D Dosages to be Added

                                40 mL                                                                                              10 mL

     Note: please convert other dosages according to equal proportion of the reagent dosage in the table

     2. Mixing the solution: after adding Solution D, cover tightly the centrifugal tube, and mix it evenly using the vortex oscillator   

     for 1 min. then let it stand at 2℃ to 8℃for at least 2 hours; (Note: adding the standing time can increase the exosome   

     attainment rate, but the standing time shall not exceed 24 hours)

     3. Precipitating exosome: take out the centrifugal tube with the mixed liquor and let it to be centrifuged at 4℃ at 10000 g for 60    

      min, discard the supernatant, and the precipitation is rich in exosomal particles;(Note: the supernatant should be sucked as    

      much as possible)

     4. Resuspending the exosome: take 1×PBS, spin the centrifugation precipitate (see the following table for specific dosage). 

     After it  is dissolved, transfer the resuspended solution into a new 1.5 mL centrifugal tube;

                                     Sample Volume                                   PBS Dosages to be Added

                                            40 mL                                                            1 mL

     Note: please convert other dosages according to equal proportion of the reagent dosage in the table

     5. Obtaining exosomal particles: centrifuge the 1.5 mL centrifugal tube with resuspended solution at 4℃ at 12000 g for 

     5 min and keep the supernatant. This supernatant is rich in exosomal particles. (Note: in case of a large amount of   

     precipitates, this step can be repeated for multiple times until there is no obvious precipitation. Collect the supernatant  

     after each centrifugation. The exosome solution may have a light milky white color, which is normal )

     6. Storing the exosome: purified exosomes shall be sub-packed at 50-100 μL and stored in a low-temperature fridge at -80℃ for  

      subsequent experimental use.

    IV Storage Conditions

     This product can be stably stored at room temperature for 24 hours. Please mix it thoroughly before use.

    V Precautions

     This product is for life science research only, and medical diagnosis or other purposes are prohibited!

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