Validation Data Gallery
Product Information
Halo-Trap Agarose are affinity beads for IP of Halo-tag proteins. It comprises a Halo-tag Nanobody /VHH conjugated to agarose beads.
| Description | The Halo-Trap Agarose Kit contains Halo-Trap Agarose, lysis, wash, and elution buffers for efficient immunoprecipitation of Halo-tagged proteins and their interacting factors.
• Halo-Trap immunoprecipitates Halo-fusion proteins even when already covalently bound to Halo-ligands, i.e. after labelling etc. • Fast, reliable & efficient one-step immunoprecipitation • No heavy & light antibody chains • Bound Halo-fusion protein can be eluted without protease • Suitable for downstream mass spec analysis |
| Applications | IP, CoIP, ChIP, RIP |
| Specificity/Target | Halo-tag (modified variant of the bacterial haloalkane dehalogenase enzyme from Rhodococcus rhodochrous) in the absence or presence of covalently bound chloralkane-based ligands. |
| Binding capacity | 7.5-10 μg of recombinant Halo-tag per 25 μL bead slurry |
| Conjugate | Agarose beads; bead size: ~ 90 µm (cross-linked 4 % agarose beads) |
| Elution buffer | SDS sample buffer 0.2 M glycine pH 2.5 |
| Wash buffer compatibility | 4 M urea, 1 M NaCl, 10 mM DTT, 2 % Nonidet P40 Substitute, 1 % Triton X-100 |
| Type | Nanobody |
| Class | Recombinant |
| Host | Alpaca |
| Affinity (KD) | Dissociation constant KD of 2 nM |
| Compatibility with mass spectrometry | The Halo-Trap is optimized for on-bead digestion. For the application note, please click here: On-bead digest protocol for mass spectrometry |
| RRID | AB_2827595 |
| Storage Buffer | 20% ethanol |
| Storage Condition | Upon receipt store at +4°C. Do not freeze! |
Kit components
| Component | Description |
|---|---|
| Halo-Trap Agarose beads | 20 reactions (500 µL) |
| Lysis buffer | Optimized for cytoplasmatic proteins and mammalian cell lysis |
| RIPA buffer | Optimized for nuclear/chromatin proteins and mammalian cell lysis |
| Wash buffer | Removal of unwanted proteins, peptides, etc. |
| Dilution buffer | Dilution of cell lysate |
| Elution buffer | For acidic elution |
Documentation
| SDS |
|---|
| otak_SDS_Halo-Trap Agarose Kit (EN) |
| Datasheet |
|---|
| Protocol Halo-Trap Agarose Kit (PDF) |
| Trouble shooting |
|---|
| Troubleshooting guide immunoprecipitation (IP) (PDF) |
Publications
| Application | Title |
|---|---|
Dev Cell The Rag GTPase Regulates the Dynamic Behavior of TSC Downstream of Both Amino Acid and Growth Factor Restriction. |