EV-ID
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.
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
| K211_2 | K211_5 | K211_10 | |
| Size | For 2 full characterisations | For 5 full characterisations | For 10 full characterisations |
| Kit content |
Internal controls (EVs and cell extract) Laemmli buffer for EV resuspension Primary antibodies (Alix, Calnexin, CD9, CD81, Syntenin-1) Secondary antibodies (HRP conjugated) |
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| Downstream application | Western Blot | ||
| Shelf life | 1 Month at -20°C | ||
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.