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Halo-Trap

3
Pull-down of Halo-tag protein
Halo-protein immunoprecipitated with either Halo-Trap (left) or an alternative supplier's product (right): Recombinantly expressed Halo-protein bound to Halo-TMR. Input (I), non-bound (FT) and bound (B) fractions were separated by SDS-PAGE followed by Coomassie blue staining, Western blotting and fluorescence scan. Please note, additional bands are visible due to sensitivity of the Halo-protein to SDS. Because the Halo-Trap binds to Halo-tag at an epitope outside its catalytic center, Halo-proteins covalently bound to ligands like Halo-TMR (left) are effectively captured. In contrast, the competitor’s product precipitates no or just minimal amounts of the Halo protein. This allows to pulldown Halo-fusion proteins from cells that have been previously treated with Halo-ligands for imaging and makes the Halo system as versatile as GFP expression, imaging and purification.
Immunoprecipitation and elution of Halo-protein from spiked HEK293T cell lysates: Input (I), non-bound (FT), eluted (E), residual (R) bound protein after boiling of beads in SDS-sample buffer and protein marker (M) were separated by SDS-PAGE followed by Coomassie blue staining and Western blotting. Elution of Halo-protein with elution buffer (low pH) from Halo-Trap is efficient (left).

Description
Halo-Trap are affinity beads for immunoprecipitation of Halo-tag fusion proteins. ChromoTek’s Halo-Trap comprises a Halo Nanobody/ VHH coupled to agarose beads.

Specificity
Halo-tag

Applications
Immunoprecipitation (IP) / Co-IP
Mass spectrometry (MS)
Enzyme activity measurements
ChIP / RIP analysis

Formats/matrices
Halo-Trap Agarose
Halo VHH, recombinant binding protein

Synonyms
anti-Halo VHH, Halo Nanobody, Halo-binding protein or anti-Halo single domain antibody (sdAb)

Benefit from Halo-Trap for Immunoprecipitation (IP)

  • Halo-Trap immunoprecipitates Halo-fusion proteins even when already covalently bound to Halo-ligands, i.e. after labelling etc.
  • Halo-Trap can be used for affinity purification
  • Bound Halo-fusion protein can be eluted without protease
  • Structure and function are characterized

Format/matrices & properties

We offer the Halo-Trap coupled to agarose beads or uncoupled as purified recombinant binding protein Halo VHH.

Extraordinary stable & reliable binding

  • Dissociation constant KD of 2 nM
  • Compatible to the following wash conditions: 4 M urea, 1 M NaCl, 10 mM DTT, 2 % Nonidet P40 Substitute, 1 % Triton X-100
  • Applications validated and characterized

Specificity

Halo-Trap binds Halo fusion proteins no matter if covalently linked to Halo ligands such as dyes, biotin, etc.

About Halo

Halo is a modified variant of a bacterial haloalkane dehalogenase enzyme from Rhodococcus rhodochrous and is designed to form a covalent bond with reactive chloroalkane-based ligands.

Find the Halo-Trap:

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Only for research applications, not for diagnostic or therapeutic use!