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PS Capture(TM) Exosome Flow Cytometry Kit

for Genetic Research
Manufacturer :
FUJIFILM Wako Pure Chemical Corporation
Storage Condition :
Keep at -20 degrees C.
GHS :
  • Structural Formula
  • Label
  • Packing
SDS
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Distributor
297-79701
Barcode No
4548995064127
300Tests
List Price
300.00 USD

In stock in Japan

Document

SDS
Product Specification Sheet
Package Insert
Spectral Data
Certificate of Analysis
Calibration Certificate

Kit component

Kit Components (300 assays)

Exosome Capture Beads 1 x 3 mL
Washing Buffer (10×) 2 x 45 mL
Exosome Binding Enhancer (100×) 1 x 15 mL

Overview

Features

  • High-Sensitive Qualitative Analysis
  • Easy Operation by Magnetic Beads
  • Direct Detection without Purification
  • Total 3 hours ~ from Isolation to Staining ~

Principle

03417241_img01.png

Procedure

Outline of Procedure ~Basic Protocol for 2 reactions~

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Flowchart of Sample Preparation

  • Cell culture supernatant
    03417241_img05.png
  • Serum・Heparin plasma
    03417241_img06.png
  • EDTA plasma and Citrated plasma
    03417241_img07R3.png

Pretreatment method of EDTA plasma and Citrated plasma

EDTA and Citric acid, which is the anticoagulant inhibit the binding of extracellular vesicles to Exosome Capture Beads. Therefore, perform pretreatment with sodium heparin solution.

  1. Prepare 1,000 U/mL sodium heparin solution with purified water. [Example of product used: Code No. 085-00134]
  2. Add 1 : 200 volume of sodium heparin solution to 100 μL of 10,000 × g supernatant (final concentration : 5 U/mL) and mix.
  3. Add 1 : 20 volume of Exosome Binding Enhancer (100×) further and mix.
  4. Proceed to “Isolation step”.

Recommended reaction scale

Basic protocol is set as 2 reactions using a 1.5 mL microcentrifuge tube to isolate EVs from samples with Exosome Capture Beads.
For scale-up, increase the amount of Exosome Capture Beads and samples. Recommended reaction scale is presented in the right table.

Note: 10 reactions are maximum for one 1.5 mL microcentrifuge tube.

Qty of Reaction Exosome Capture Beads (µL) Sample Volume(µL)
2 reactions
(basic)
30 100
3 reactions 40 133
4 reactions 50 167
5 reactions 60 200
6 reactions 70 233
7 reactions 80 267
8 reactions 90 300
9 reactions 100 333
10 reactions 110 367

Applications

Qualitative analysis of exosomes in cell culture supernatant of K562 cell line

Exosomes in cell culture supernatant of K562 cells were isolated using PS Capture™ Exosome Flow Cytometry Kit or each of anti-CD81-, CD9- and CD63-antibody-immobilized magnetic beads (supplier A), followed by flow cytometric analysis of exosome surface antigens after immunostaining with fluorescence-labeled antibodies.

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Sample Cell culture supernatant of K562 cells: 33 μL/Assay
Detection antibody PE-anti-CD63 (BD Biosciences, 556020)
PE-anti-CD9 (Novus Biologicals, NB100-77915PE)
PE-anti-CD81 (Novus Biologicals, NBP1-44861PE)

03417241_img03.png

① PS Capture™ Exosome Flow Cytometry Kit
② Anti-CD81 antibody immobilized magnetic beads
③ Anti-CD9 antibody immobilized magnetic beads
④ Anti-CD63 antibody immobilized magnetic beads


Each signal value was normalized using the value of Isotype.

It was confirmed that the PS Capture™ Exosome FCM Kit can detect the exosome surface antigen with high sensitivity compared with competitors' products, regardless of which detection antibody is used.

Qualitative analysis of exosomes in serum and plasma

Exosomes in human serum and human plasma (EDTA plasma, heparin plasma) were isolated using PS Capture™ Exosome Flow Cytometry Kit, followed by flow cytometric analysis of exosome surface antigens after immunostaining with PE-labeled mouse IgG isotype control and PE-labeled anti-human CD9 antibody.

Sample:
33 μL/Assay each
Human serum
Human heparin plasma
Human EDTA plasma (buffer exchanged)
Detection antibody PE-labeled anti-CD9 antibody
(Novus Biologicals, NB100-77915PE)
03417241_img04.png

In any of the samples, the peak shift of fluorescence intensity was confirmed when stained with PE-labeled anti-human CD9 antibody.

Overview / Applications

Outline This kit is a reagent that can qualitatively analyze extracellular vesicles contained in cell culture supernatants and body fluid samples by flow cytometry. After extracellular vesicles are reacted and immobilized on magnetic beads on which extracellular vesicle surface phosphatidylserine (PS) specifically binding protein is immobilized, fluorescent labeling for any extracellular vesicle marker protein by using antibodies, we can detect marker proteins on extracellular vesicle surface with high sensitivity.
Before using it, prepare the primary antibody for each surface marker protein, the fluorescent dye-labeled secondary antibody, or the primary antibody labeled with fluorescent dye.

Property

Manufacturer Information

Alias

For research use or further manufacturing use only. Not for use in diagnostic procedures.

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