Validation Data Gallery
Product Information
Kit for IP/Co-IP of HA-fusion proteins and sample preparations for mass spectrometry (MS)
| Description | The iST HA-Trap Kit enables researchers to process HA-fusion proteins and their interacting partners for mass spectrometry analysis by including the ChromoTek HA-Trap for IP /Co-IP of HA-fusion 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 accomodates up to eight samples and includes pull-down material for 2 controls. |
| Applications | IP, Co-IP, MS |
| Specificity/Target | Binds specifically to the HA-tag (sequence YPYDVPDYA) fused to a protein of interest at N-, C- or internal position. Please note that the affinity is highest for a C-terminal fusion. There is no cross-reactivity to other common peptide tags such as the His6-tag, FLAG-tag, Spot-Tag, V5-tag, Strep-tag, or C-tag (other tags not tested). Background binding to host cell proteins from a range of organisms such as human, mouse and dog cell lines or yeast and plants is low. |
| Binding Capacity | 20 ug of recombinant HA-tagged protein (~30 kDa) per 25 uL bead slurry |
| Conjugate | Agarose beads; ~90 um (cross-linked 4% agarose beads) |
| Elution buffer | 2x SDS-sample buffer (Lammli) |
| Type | Nanobody |
| Class | Recombinant |
| Host | Alpaca |
| Affinity (KD) | 6 nM for C-terminal HA-tags and ca. 180 nM for N-terminal fusions. |
| Compatibility with mass spectrometry | The HA-Trap is optimized for on-bead digestion. |
| Storage Buffer | 20% ethanol |
| Storage Condition | Shipped at ambient temperature. Upon receipt store HA-Trap at +4°C and Digest at -20°C. |
Kit components
| Component | Description |
|---|---|
| HA-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 hydrophillic contaminants |
| Elution buffer | Elute peptides from the cartridge |
| LC-Load | Load peptides on reversed-phase LC-MS column |
| Cartridges | Cartridge for 1 to 100 ug 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 |
|---|
| iST HA-Trap IP-MS Sample Preparation Kit Datasheet |