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- Regulatory Status
- RUO
- Other Names
- Microbead Human CD81 Exosome Selection Kit, Exosome positive isolation, Exosome enrichment
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- Product Citations
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Following the kit protocol, human CD81 exosomes were positively selected using the MojoSort™ Human CD81 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) (Cat. No. 406401) at a 1:2000 dilution.
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Figure 1. Western Blot Analysis of isolated exosomes using MojoSort™ Human CD81 Microbead Exosome Kit.
Following the kit protocol, human CD81 exosomes were positively selected using the MojoSort™ Human CD81 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) (Cat. No. 406401) at a 1:2000 dilution. -
Figure 2. Western Blot Analysis of isolated exosomes using MojoSort™ Human CD81 Microbead Exosome Kit.
Following the kit protocol, human CD81 exosomes were positively selected using the MojoSort™ Human CD81 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) (Cat. No. 406401) at a 1:2000 dilution. -
Figure 3. Western Blot Analysis of isolated exosomes using MojoSort™ Human CD81 Microbead Exosome Kit.
Following the kit protocol, human CD81 exosomes were positively selected using the MojoSort™ Human CD81 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) (Cat. No. 406401) at a 1:2000 dilution. -
Figure 4. Western Blot Analysis of isolated exosomes using MojoSort™ Human CD81 Microbead Exosome Kit.
Following the kit protocol, human CD81 exosomes were positively selected using the MojoSort™ Human CD81 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.
The MojoSort™ Human CD81 Microbead Exosome Kit is intended for the positive selection of human CD81 exosomes from cell culture supernatant, plasma, serum, or urine. MojoSort™ Microbeads are directly conjugated to anti-CD81 mouse monoclonal antibodies. MojoSort™ Microbeads are superparamagnetic particles with an average diameter of 1 μm. Tetraspanin membrane proteins, such as CD81, are enriched on the surface of exosomes and can be used for separation from biological samples. The positively selected bead-bound CD81 exosomes may be characterized by downstream applications such as western blot, flow cytometry, and qRT-PCR. The bead-bound CD81 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.
Kit Contents
- Kit Contents
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• 1 vial of MojoSort™ Human CD81 Exosome Microbeads (900 μL)
• 1 bottle of MojoSort™ Exosome Buffer (10X) (25 mL)
Product Details
- Verified Reactivity
- Human
- Formulation
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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
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Purification - Quality tested
- Recommended Usage
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20 μL of Microbeads per 0.5 mL human biofluids
- Application Notes
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Kit Capacity:
This kit provides 900 μL of MojoSort™ Human CD81 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, cell culture supernatant, or urine). Per test, we recommend selection using 20 μL of MojoSort™ Human CD81 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 CD81 tetraspanins on the surface of exosomes may vary between source of biofluids (serum, cell culture supernatant, or urine) and may also be donor dependent. Varying CD81 levels on human exosomes may also affect yield and require titration of kit components.
Antibody or cocktail dilution to use in column: N/A
Dilution to use in column: N/A - 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.
Antigen Details
- Molecular Family
- Exosome Markers
- Gene ID
- NA
- UniProt
- View information about CD81 on UniProt.org
Related FAQs
- Is there a way to detach your magnetic particles from the cell surface?
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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?
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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?
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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?
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Please contact our technical service team for further assistance.
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