Validation Data Gallery
Product Information
Spot-Trap® Agarose is an affinity bead for immunoprecipitation (IP) of Spot-tagged proteins It comprises an anti-Spot-Tag® VHH/ Nanobody coupled to agarose beads.
| Description | Immunoprecipitation of Spot-tagged proteins and their interacting factors with anti-Spot Nanobody conjugated to agarose beads.
• No contaminating heavy and light antibody chains • Ready-to-use • Recombinantly expressed without batch-to-batch variation • Well-characterized and validated • High stability allows for stringent washing • Extraordinary low background compared to other affinity resins • 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 | Agarose beads; bead size: ~ 90 µm (cross-linked 4 % 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_2827590 |
| 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. |
| Size | 25ul/reactions (eg：20rxns=500ul slurry) |
Documentation
| SDS |
|---|
| eta_SDS_Spot-Trap® Agarose (EN) |
| Datasheet |
|---|
| Spot-Trap® Agarose Datasheet |
| Poster |
|---|
| Scientific Spot system poster |
| Application notes |
|---|
| Applications note Spot-System for for CRISPR’ed Spot-Tag |
| Troubleshooting |
|---|
| Troubleshooting guide immunoprecipitation (PDF) |
Publications
| Application | Title |
|---|---|
J Biol Chem Serine phosphorylation of the small phosphoprotein ICAP1 inhibits its nuclear accumulation. | |
Biomolecules Nanobodies Right in the Middle: Intrabodies as Toolbox to Visualize and Modulate Antigens in the Living Cell. | |
J Cell Sci 30 years of nanobodies - an ongoing success story of small binders in biological research | |
J Cell Sci Identification of distinct active pools of yeast serine palmitoyltransferase in sub-compartments of the ER | |
EMBO J Jacob-induced transcriptional inactivation of CREB promotes Aβ-induced synapse loss in Alzheimer's disease |