Validation Data Gallery
Product Information
Kit for IP/ Co-IP of V5-tagged proteins & sample preparation for mass spectrometry (MS)
| Description | The iST V5-Trap® Kit enables researchers to process V5-tagged proteins and their interacting partners for Mass Spectrometry analysis by including the ChromoTek V5-Trap for immunoprecipitation/Co-IP of V5-tagged proteins and the PreOmics iST buffers and cartridges required for bottom-up proteomic sample preparation. This robust method yields purified peptides while dramatically reducing contamination and sample loss. Each kit accommodates up to eight samples and includes pull-down reagent for controls:
• Reliable identification of V5-tagged proteins & interacting partners by mass spec • Superior processing of IP samples into clean peptide mixtures • Intuitive protocol and reproducible results |
| Applications | IP, MS |
| Specificity/Target | V5-tag sequence GKPIPNPLLGLDST at the N-terminus, C-terminus, or internal site of the fusion protein. |
| Binding capacity | 15 μg of recombinant V5-tagged protein (~30 kDa) per 25 μL bead slurry |
| Conjugate | Agarose beads; bead size: ~ 90 µm (cross-linked 4 % agarose beads) |
| Type | Nanobody |
| Class | Recombinant |
| Host | Alpaca |
| Affinity (KD) | Dissociation constant KD of 40 nM |
| Compatibility with mass spectrometry | The V5-Trap® is optimized for on-bead digestion. |
| Storage Buffer | 20% ethanol |
| Storage Condition | Kit is shipped at ambient temperature. Upon receipt store V5-Trap at 4°C and Digest at -20°C |
Kit components
| Component | Description |
|---|---|
| V5-Trap™ Agarose | 8 reactions plus 2 controls |
| Digest | Enzyme Trypsin-mix to digest proteins. |
| Resuspend buffer | Protease reconstitution buffer for enzymes. |
| Lyse buffer | Denature, reduce and alkylate proteins. |
| Stop solution | Stop the enzymatic activity. |
| Wash 1 buffer | Clean up peptides from hydrophobic contaminants. |
| Wash 2 buffer | Clean up peptides from hydrophilic contaminants. |
| Elution buffer | Elute the peptides from the cartridge. |
| LC-Load load | Load peptides on reversed-phase LC-MS column. |
| Cartridges | Cartridge for 1 to 100 µg protein starting material. |
| Waste tubes | Tube for collecting waste after washing steps. |
| Collection tubes | Tube for collecting peptides after elution. |
| Adapters | Enables placing a cartridge into a tube. |
| Caps | Cap to optionally close the cartridge’s bottom. |
Documentation
| SDS |
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| SDS iST Kits (grouped) EtOH (EN)-CTK |
| Datasheet |
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| Protocol v5tak-iST-iST V5-Trap Kit (PDF) |