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MojoSort™ Human CD63 Microbead Exosome Kit

Pricing & Availability
Regulatory Status
RUO
Other Names
Human CD63 positive selection, Exosome positive selection, Exosome positive isolation
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Human-CD63-Microbead-Exosome-Kit_1_111824
Figure 1. Western Blot Analysis of isolated exosomes using MojoSort™ Human CD63 Microbead Exosome Kit. Following the kit protocol, human CD63 exosomes were positively selected using the MojoSort™ Human CD63 Microbead Exosome Kit from 500 μL of conditioned HEK293T cell culture medium. For comparison, exosomes were also isolated using two competitor kits following the kit-specific protocols. The three exosome samples collected using the three different kits were lysed for western blot. Equal volumes of reduced lysate were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane and probed with a rabbit monoclonal anti-Alix antibody overnight. Proteins were visualized by chemiluminescence detection using HRP Donkey anti-rabbit IgG (minimal x-reactivity) Antibody (Cat. No. 406401) at a 1:2000 dilution.
  • Human-CD63-Microbead-Exosome-Kit_1_111824
    Figure 1. Western Blot Analysis of isolated exosomes using MojoSort™ Human CD63 Microbead Exosome Kit. Following the kit protocol, human CD63 exosomes were positively selected using the MojoSort™ Human CD63 Microbead Exosome Kit from 500 μL of conditioned HEK293T cell culture medium. For comparison, exosomes were also isolated using two competitor kits following the kit-specific protocols. The three exosome samples collected using the three different kits were lysed for western blot. Equal volumes of reduced lysate were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane and probed with a rabbit monoclonal anti-Alix antibody overnight. Proteins were visualized by chemiluminescence detection using HRP Donkey anti-rabbit IgG (minimal x-reactivity) Antibody (Cat. No. 406401) at a 1:2000 dilution.
  • Human-CD63-Microbead-Exosome-Kit_2_111824
    Figure 2. Western Blot Analysis of isolated exosomes using MojoSort™ Human CD63 Microbead Exosome Kit. Following the kit protocol, human CD9 exosomes were positively selected using the MojoSort™ Human CD63 Microbead Exosome Kit from 500 μL of conditioned HEK293T cell culture medium. For comparison, exosomes were also isolated using two competitor kits following the kit-specific protocols. The three exosome samples collected using the three different kits were lysed for western blot. Equal volumes of reduced lysate were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane and probed with a rabbit monoclonal anti-CD9 antibody overnight. Proteins were visualized by chemiluminescence detection using HRP Donkey anti-rabbit IgG (minimal x-reactivity) Antibody (Cat. No. 406401) at a 1:2000 dilution.
  • Human-CD63-Microbead-Exosome-Kit_3_111824
    Figure 3. Western Blot Analysis of isolated exosomes using MojoSort™ Human CD63 Microbead Exosome Kit. Following the kit protocol, human CD63 exosomes were positively selected using the MojoSort™ Human CD63 Microbead Exosome Kit from 500 μL of conditioned HEK293T cell culture medium. For comparison, exosomes were also isolated using two competitor kits following the kit-specific protocols. The three exosome samples collected using the three different kits were lysed for western blot. Equal volumes of reduced lysate were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane and probed with a rabbit monoclonal anti-CD63 antibody overnight. Proteins were visualized by chemiluminescence detection using HRP Donkey anti-rabbit IgG (minimal x-reactivity) Antibody (Cat. No. 406401) at a 1:2000 dilution.
  • Human-CD63-Microbead-Exosome-Kit_4_111824
    Figure 4. Western Blot Analysis of isolated exosomes using MojoSort™ Human CD63 Microbead Exosome Kit. Following the kit protocol, human CD63 exosomes were positively selected using the MojoSort™ Human CD63 Microbead Exosome Kit from 500 μL of conditioned HEK293T cell culture medium. For comparison, exosomes were also isolated using two competitor kits following the kit-specific protocols. The three exosome samples collected using the three different kits were lysed for western blot. Equal volumes of reduced lysate were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane and probed with a rabbit monoclonal anti-CD81 antibody overnight. Proteins were visualized by chemiluminescence detection using HRP Donkey anti-rabbit IgG (minimal x-reactivity) (Cat. No. 406401) at a 1:2000 dilution.
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Description

The MojoSort™ Human CD63 Microbead Exosome Kit is intended for the magnetic separation of human CD63 exosomes from cell culture supernatant, heparin-treated plasma, serum or urine. MojoSort™ Microbeads are directly conjugated to anti-CD63 mouse monoclonal antibodies. MojoSort™ Microbeads are superparamagnetic particles with an average diameter of 1 μm. Tetraspanin membrane proteins, such as CD63, are enriched on the surface of exosomes and can be used for separation from biological samples. The positively selected bead-bound CD63 exosomes may be characterized by downstream applications such as western blot, flow cytometry and qRT-PCR. The bead-bound CD63 exosomes may also be released from the beads under acidic conditions (see “Elution of Exosomes” section in the protocol).

Unlike other exosome isolation kits in the market, this kit does not require users to pre-enrich their samples with ultracentrifugation or total exosome enrichment kits. MojoSort™ reagents are also compatible with magnets from other vendors. Optimizing incubations times when using magnets from other vendors is recommended. Please contact Technical Services for more information at https://www.biolegend.com/en-us/contact 

Product Details
Technical Data Sheet (pdf)

Kit Contents

Kit Contents
  • 1 vial of MojoSort™ Human CD63 Exosome Microbeads (900µL)
  • 1 bottle of MojoSort™ Exosome Buffer (10X) (25mL)

Product Details

Verified Reactivity
Human
Formulation
Particle: Aqueous solution containing sodium phosphate
Buffer: Aqueous solution
Preparation
The antibodies were purified by affinity chromatography and conjugated onto the magnetic carboxyl microbeads.
Storage & Handling
All components should be stored upright and undiluted between 2°C and 8°C.
Application

Purification – Quality tested
Recommended Usage

20 μL of Microbeads per 0.5 mL human biofluids (serum, heparin-treated plasma, cell culture supernatant or urine)

Application Notes

Kit Capacity:

This kit provides 900 μL of MojoSort™ Human CD63 Exosome Microbeads and 25 mL of MojoSort™ Exosome Buffer (10X). For use of this kit, a single test is defined as processing 0.5 mL of human biofluids (serum, heparin-treated plasma, cell culture supernatant or urine). Per test, we recommend separation using 20 μL of MojoSort™ Human CD63 Exosome Microbeads. This kit provides enough reagents to process approximately 20 mL of human biofluids.

To obtain more MojoSort™ Exosome Buffer (10X), please contact Technical Services for more information at https://www.biolegend.com/en-us/contact.

Starting sample notes:

  • If you intend to select exosomes from < 0.5 mL of biofluids per test, please bring up the volume to 0.5 mL using 1X Exosome Buffer and adhere to indicated volumes for 0.5 mL tests according to the protocol. Please note that using less than the recommended volume of 0.5 mL of starting sample may result in reduced exosome yield.
     
  • Levels of CD63 tetraspanins on the surface of exosomes may vary between source of biofluids (serum, heparin-treated plasma, cell culture supernatant or urine) and may also be donor dependent. Varying CD63 levels on human exosomes may also affect yield and require titration of kit components.
     
  • We may see a faint non-specific band around 27 kD and 55 kD; this is due to the light chain being separated from the heavy chain (vice versa) and falling off from the antibody conjugated beads while lysing and preparing the bead bound exosomes for running in SDS gel in reduced form (because of denaturation with SDS in the sample buffer and reduction with beta-mercaptoethanol).
     
  • This kit is validated for use with Heparin-treated plasma only. It is not suitable for other types of plasma.

Antibody or cocktail dilution to use in column: N/A 
Dilution to use in column: N/A 

Related FAQs

Is there a way to detach your magnetic particles from the cell surface?

MojoSort™ Nanobeads: These magnetic particles cannot be removed from cells. This includes standalone Nanobeads and MojoSort kits using Nanobeads. We have found that cells are functional without the need to detach the magnetic Nanobeads.

 

MojoSort Microbeads: These magnetic particles may be released following the detailed product-specific protocol.

What is the size of your magnetic particles?

MojoSort Nanobeads: the average diameter is approximately 130 nm.

 

MojoSort Microbeads: the average diameter is approximately 1 μm.

Are MojoSort™ Nanobeads compatible with other commercially available magnetic separation systems?

MojoSort™ magnetic particles can be used with other commercially available magnetic separators, both free standing magnets and column-based systems. Because MojoSort™ protocols are optimized for the MojoSort™ separator, the protocols may need to be adjusted for other systems.  Please contact BioLegend Technical Service for more product-specific information and guidance. We do not recommend using MojoSort™ particles for BD’s IMag™ or Life Technologies’ DynaMag™.

What antibodies are present in the depletion cocktails provided for isolation kits?

Please contact our technical service team for further assistance.

Go To Top Version: 1    Revision Date: 11.18.2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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