Validation Data Gallery
Product Information
Spot-Trap® Magnetic Agarose is an affinity reagent for immunoprecipitation (IP) of Spot-tagged proteins. It consists of an anti-Spot-Tag® VHH/ Nanobody that is coupled to magnetic agarose beads.
| Description | The Spot-Trap Magnetic Agarose Kit contains Spot-Trap Magnetic Agarose, lysis, wash, and elution buffers for efficient immunoprecipitation of Spot-tagged proteins and their interacting factors.
• No contaminating heavy and light antibody chains • Ready-to-use • Recombinantly expressed without batch-to-batch variation • Extraordinary low background compared to other affinity resins • Compatible with harsh buffer conditions • Suitable for downstream mass spec analysis • Short incubation time of 30-60 minutes |
| Applications | IP, CoIP, ChIP, RIP |
| Specificity/Target | Spot-Tag® sequence PDRVRAVSHWSS |
| Binding capacity | 17.5 μg of recombinant Spot-tagged protein (~30 kDa) per 25 μL bead slurry |
| Conjugate | Magnetic agarose beads; bead size: ~40 µm (cross-linked 6 % magnetic agarose beads) |
| Elution buffer | SDS sample buffer 10 mM NaOH pH 12 |
| Wash buffer compatibility | 2 M urea, 2 M NaCl, 10 mM DTT, 2 % Nonidet P40 Substitute, 1 % Triton X-100, 0.1 % SDS |
| 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. For the application note, please click here: On-bead digest protocol for mass spectrometry |
| RRID | AB_2827591 |
| Storage Buffer | 20% ethanol |
| Storage Condition | Shipped at ambient temperature. Upon receipt store at 4°C. Stable for one year. Do not freeze! |
| 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® Magnetic Agarose | 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 |
Documentation
| SDS |
|---|
| etmak_SDS_Spot-Trap® Magnetic Agarose Kit (EN) |
| Datasheet |
|---|
| Protocol Spot-Trap Magnetic Agarose Kit (PDF) |
| Application notes |
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| Applications note Spot-System for for CRISPR’ed Spot-Tag |
| Troubleshooting |
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| Troubleshooting guide immunoprecipitation (PDF) |