Validation Data Gallery
Product Information
Kit for IP/ Co-IP of Spot-tagged proteins & sample preparation for mass spectrometry (MS)
| Description | The iST Spot-Trap® Kit enables researchers to process Spot-tagged proteins and their interacting partners for Mass Spectrometric analysis. The kit includes the ChromoTek Spot-Trap for immunoprecipitation/Co-IP of Spot-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 dentification of Spot-tagged proteins & interacting partners by mass spectrometry. • Identification of even low abundant/ low expressed Spot-tagged proteins • Superior processing of IP samples into clean peptide mixtures • Intuitive protocol and reproducible results |
| Applications | IP, MS |
| Specificity/Target | Spot-Tag® sequence PDRVRAVSHWSS |
| Binding capacity | 17.5 μg of recombinant Spot-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 6 nM |
| Compatibility with mass spectrometry | The Spot-Trap® is optimized for on-bead digestion. |
| Storage Buffer | 20% ethanol |
| Storage Condition | Kit is shipped at ambient temperature. Upon receipt store Spot-Trap at 4°C and Digest at -20°C |
| Spot-Tag® Origin | Engineered 12 amino acid sequence modified from the unstructured N-terminus of beta-catenin |
| Cross Reactivity | The Spot-Nanobody shows minimal cross reactivity with beta-catenin. |
Kit components
| Component | Description |
|---|---|
| Spot-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 |
|---|
| SDS iST Kits (grouped) EtOH (EN)-CTK |
| Datasheet |
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| Protocol etak-iST-iST Spot-Trap Kit (PDF) |