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Exosome Isolation Tools, EXO-Prep Reagents

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Exosome Isolation Tools, EXO-Prep Reagents

Exosomes one step isolation from a wide range of biofluids(plasam, serum & urine) or cell culture

supernatants. 

 

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Isolate exosome from your sample by EXO-Prep

EXO-Prep is a fast and convenient method of exosome isolation from biofluids and cell culture supernatants.

EXO-Prep is based on chemical precipitation. Samples are incubated with EXOPrep solution on ice, so that

exosomes will precipitate following centrifugation. The obtained pellet can be resuspended in PBS 1X or

deionized water. The protocol is user friendly, time-saving (around 1 hour), and does not requirecapital

laboratory equipment. Isolated exosomes are suitable for a wide range of analyses, such as NTA, protein

profiling by using different techniques (western blotting, ELISA, FACS), nucleic acid extraction and profiling of

mRNA or small RNA markers. 

 

Exo-Prep, working procedure and potential applications 

 

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Fig. 1. Working procedure

 

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Fig. 2. ELISA assay. Exosomes isolated by EXO-Prep are suitable for protein profiling using ELISA assay.

ELISA performed on HBM immunoplate for overall exosome capture from human plasma (HBM-POF)

 

 

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Fig. 3. Western blotting. Detection of Exosome marker Alix in

protein lysates (30 ug) from exosomes isolated with EXO-Prep

from 100 ul and 500 ul of plasma/serum and 10 and 20 ml of

whole urine. Protein lysates (30 ug) from exosomes purified by

ultracentrifugation (UC) were used as control.

 

 

 

 

 

 

 

 

 

 

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Fig. 4. Nucleic acids extraction and analysis. Exosomes previously isolated by EXO-Prep or ultracentrifugation

(UC) from 100 ul or 500 ul of plasma were used for total RNA extraction and profiling of four exosomal miRNAs.

Exosome isolated with EXO-Prep are enriched in miRNAs compared to those purified by ultracentrifugation

from the same amount of plasma. 

 

                                   Exoxome isolated from 100 μl of plasma

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Fig. 5. Nano Tracking Analysis (NTA). NTA performed on exosomes isolated by EXO-Prep and resuspended

in 100 ul of PBS 1X. Size of particles is in according to the size of exosome vesicles (50-120 nm) 

 

NTA analysis performed on the same plasma sample before and after EXO-Prep treatment shows that EXO-

Prep is able to isolate almost the entire exosome population in the sample. Indeed the number of particles in

Exosome isolated by EXO-Prep ( 8.67x10^11) corresponds to that present in whole plasma (8.92x10^11).

Viceversa plasma after EXO-Prep treatment remains depleted of microvesicles (0.91x10^11).

 

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Fig. 6. NTA and exosome detection in whole plasma (A), EXO-Prep isolated exosomes resuspended in 100 ul

of PBS 1x (B) and plasma post EXO-Prep treatment (C).

  

Applications

Isolated exosomes are suitable for,

• Proteomic analyses (ELISA, WB)

• Nucleic acid extraction and analyses

• Nano Tracking analysis (Nanosight)

• Flow cytometry

• Electron microscopy

 

Advantages

• Time and money saving

• Easy and fast procedure

• No ultracentrifugation required

• Thrombin pretreatment of plasma not required

• Allows exosome isolation from biofluids and cell culture supernatants

• Small sample volume required (as low as 100 ul of plasma)

• Suitable for exosomal nucleic acid(including miRNA) extraction and protein profiling

• Easy to store and ship (4℃)

 

 

EXO-Prep guarantees exosome isolation form complex biofluids, high performances

and cheaper price than competitor products

 

EXO-Prep is a fast and efficient method for exosome isolation from biofluids or cell culture media. In particular,

when compared to a competitor product on the market, EXO-Prep allows exosome isolation from a smaller

volume (100 ul) of complex biofluids as plasma, without thrombin pretreatment or difficult solubilization of

exosome pellet. EXO-Prep-isolated exosomes are suitable for different analyses: proteinprofiling via western

blotting or ELISA (Fig 7), nucleic acids extraction and miRNA/mRNA profiling (Fig 8), FACS. 

 

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Fig. 7. CD9 expression of plasma exosomes from two healthy individuals (100 μl and 250 μl) using EXO-Prep

(EP) isolation reagent compared to a competitor reagent (Comp) and exosomes purified by ultracentrifugation

(UC).

  

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Fig. 8. RNA extraction and profiling of 4 exosome miRNA from exosomes isolated by EXO-Prep, a Competitor

reagent and ultracentrifugation (UC). 100 μl of plasma sample have been used.

 

 

Comparison table

   

Descriptions

EXO-Prep

Competitor

  Exosome isolation from cell supernatants

Yes

Yes

  Exosome isolation from complex biofluids

  (plasma, serum, urine)

Yes

Yes, but pellet is hard

to solubilize

  Plasma exosome isolation

  Thrombin pretreatment

Not required

Necessary

  Pellet solubility

Easy

Very hard

  Minimum sample amount

100 μl

250 μl

  Isolated exosome are intact and suitable for ELISA assay

Yes

No suitable for ELISA assay

  Suitable for nucleic acids extraction and analysis

Yes

Yes

  Price (for 5 ml of reagent)

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Ordering informations

    

- Exosome one step isolation from plasma & serum

Catalog No.

Product Name

Size

HBM-EXP-B5

EXO-Prep, 180 reactions Plasma, 80 reactions Serum

5 ml

HBM-EXP-B10

EXO-Prep, 350 reactions Plasma, 160 reactions Serum

10 ml

HBM-EXP-B20

EXO-Prep, 700 reactions Plasma, 320 reactions Serum

20 ml

 

- Exosome isolation from Cell Supernatants

Catalog No.

Product Name

Size

HBM-EXP-C25

EXO-Prep, 25 reactions

25 ml

HBM-EXP-C50

EXO-Prep, 50 reactions

50 ml

 

- Exosome one step isolation from urine

Catalog No.

Product Name

Size

HBM-EXP-U25

EXO-Prep, 25 reactions

30 ml

HBM-EXP-U50

EXO-Prep, 50 reactions

60 ml

 

 

▣ 관련 페이지 ; HansaBioMed 

 

   

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