EV-ID
Multi-Species EV characterisation kit for Western blot analysis
Reference K211
EV-ID kit allows to characterise EVs by Western blot for the presence of EV positive markers and the absence of cellular contamination marker.
EV-ID kit allows to characterise EVs by detecting specific EV markers and a cellular contamination marker by Western blot analysis. Indeed, due to the heterogeneous nature of EVs, as well as the complexity of biological samples, a characterisation of EVs after isolation is recommended to confirm that the isolated particles are EVs and not products of cell fragmentation or other contaminants, usually present in biological samples.
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To confirm the presence of EVs, EV-ID kit allows the detection of 2 surface EV markers, CD9 and CD81 and 2 internal EV markers, Alix and Syntenin-1. Moreover, EV-ID kit includes the detection of Calnexin, a cellular contamination marker, that must be absent in the pure EV preparations.
Schematic representation of an EV, harbouring the different markers revealed by Western blot using EV-ID kit.
Created using biorender.com
Specifications
EV-ID kit is designed for the characterisation of semi or highly purified EVs from both human and animal origins.
It is necessary to use a sufficient amount of EVs allowing to detect EV markers by SDS-PAGE and Western blot.
Strengths of EV-ID kit
o   Easy to use
o   Quick
o   Reliable
Additional information
| Â | K211_2 | K211_5 | K211_10 |
|  Size | For 2 full characterisations | For 5 full characterisations | For 10 full characterisations |
| Â Kit content |
Internal controls (EV lysate and cell extract) Laemmli buffer for EV resuspension Primary antibodies (Alix, Calnexin, CD9, CD81, Syntenin-1) Secondary antibodies (HRP conjugated) |
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| Â Downstream application | Western Blot | ||
| Â Shelf life | 6 months | ||
Data and examples
An EV Batch was produced and purified according to in-EV process. This EV batch was characterised using EV-ID kit. The presence of EV surface markers (CD9 and CD81), EV luminal markers (Alix and Syntenin-1) and Calnexin, a cellular contamination marker was assessed. Control EVs (EV CTRL +) and control cellular extract (CELL CTRL +), provided in the kit, were used.
Characterisation of pure EVs by Western blot using EV-ID kit.
The EV markers were detected in the EV batch, attesting the presence of EVs. No Calnexin was detected in the EV batch, showing the absence of cellular contaminant.Â