Efficient immunoprecipitation & purification of Spot-tagged proteins
ChromoTek Spot-Trap is a high quality Spot-Tag® binding protein coupled to agarose beads or magnetic agarose beads. The Spot-tag binding protein, also termed anti-Spot-Tag nanobody or anti-Spot-Tag VHH, is derived from heavy chain antibodies. The Spot-Trap is an affinity resin for fast and effective immunoprecipitation and affinity purification. It is optimized for biochemical analysis of Spot-tagged proteins and their interacting partners. The Spot-Trap is well-characterized and validated.
Use Spot-Trap for:
- Immunoprecipitation (IP) / Co-IP
- Immunoprecipitation (IP) / Co-IP for Mass spec analysis
- Affinity purification
- Enzyme activity measurements
Spot-Tag is an inert 12 amino acid peptide tag with the sequence PDRVRAVSHWSS. Please find more information about the Spot-Tag here.
Spot-Trap is a very low background affinity reagent
See benchmark of Spot-Trap Agarose and Magnetic Agarose with other suppliers’ capture tools. Immunoprecipitations were conducted from HEK293T cell lysates without Spot-Tag present and normalized for equal binding capacity. The IPs were done according to respective manufacturers’ protocols. It is apparent that Spot-Trap does not only lack heavy and light chain antibody contaminations but also unspecific binding. This makes Spot-Trap a superior affinity capture tool for single band purification of Spot-tagged proteins.
Try before you buy: Get a test sample of the Spot-Trap® - in return we just need you to send back your test results within 4 weeks.
- NO heavy & light antibody chains
- High binding affinity
- Harsh washing conditions
- Short incubation time of 30-60 minutes
Pulldown of Spot-tagged protein. Input (I), non-bound (FT) and bound (B) fractions were separated by SDS-PAGE followed by Coomassie staining and Western blotting. Western blot indicates high effectiveness of pull-down.
- High affinity
- Purification of low abundant proteins
- Low background, pure results
- Native and non-native elution
Elution of affinty purified, Spot-tagged protein. Input (I), non-bound (FT) and eluted (E1, E2 and E3) fractions were separated by SDS-PAGE followed by Coomassie staining and Western blotting. Elutions with Spot-peptide. Western blot indicates effective elution.
Regeneration of Spot-Trap beads is possible. It is recommended, however, to re-use beads for the repeated purification of the same protein only to avoid cross-contamination.
Available matrices for Spot-Trap
|Spot-binding protein coupled to||agarose beads||magnetic agarose beads|
|binding capacity for a 30kDa protein (resin)||1.4µg/µl|
|centrifuged up to||3000g||3000g|
In addition, we offer non-conjugated Spot-Tag binding protein (=Spot-Nanobody):
- Spot-Label for Western blot (WB), uncoupled