Validation Data Gallery
![V5-Trap® Agarose for immunoprecipitation of V5 fusion proteins. HEK293T cell lysate with V5-tagged protein. Coomassie and Western blot. V5-tag antibody [SV5-P-K], monoclonal mouse IgG1 kappa and Alpaca anti-mouse IgG1, Fc-specific recombinant VHH, Alexa Fluor® 488 I: Input, FT: Flow-Through, B: Bound.](https://www.ptglab.com/products/pictures/Result-v5tak_20220118100647702.png "V5-Trap® Agarose for immunoprecipitation of V5 fusion proteins. HEK293T cell lysate with V5-tagged protein. Coomassie and Western blot. V5-tag antibody [SV5-P-K], monoclonal mouse IgG1 kappa and Alpaca anti-mouse IgG1, Fc-specific recombinant VHH, Alexa Fluor® 488 I: Input, FT: Flow-Through, B: Bound.")
Product Information
V5-Trap® Agarose is an affinity reagent for IP and purification of V5-tagged proteins. It consists of an anti-V5 Nanobody/ VHH coupled to agarose beads.
| Description | The V5-Trap Agarose Kit contains V5-Trap Agarose, lysis, wash, and elution buffers for efficient immunoprecipitation of V5-tagged proteins and their interacting factors.
• Fast, reliable & efficient one-step immunoprecipitation • Effective elution with V5-peptide • No heavy & light antibody chains on the SDS-PAGE • Stable under harsh washing conditions • Suitable for downstream mass spec analysis |
| Applications | IP, CoIP, ChIP, RIP, Protein purification |
| 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) |
| Elution buffer | V5-peptide SDS sample buffer 0.2 M glycine pH 2.5 |
| Wash buffer compatibility | 2 % Nonidet P40 Substitute, 2 % Triton X-100, 1 M NaCl, 5 mM TCEP, 5 mM DTT, 10 mM ß-mercaptoethanol, 4 M Urea |
| 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. For the application note, please click here: On-bead digest protocol for mass spectrometry |
| RRID | AB_2868497 |
| Storage Buffer | 20% ethanol |
| Storage Condition | Shipped at ambient temperature. Upon receipt store at 4°C. Stable for one year. Do not freeze! |
Kit components
| Component | Description |
|---|---|
| V5-Trap™ 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 |
|---|
| v5tak_SDS_V5-Trap® Agarose Kit (EN) |
| Datasheet |
|---|
| Protocol V5-Trap Agarose Kit (PDF) |
| Troubleshooting |
|---|
| Troubleshooting guide immunoprecipitation (PDF) |
Publications
| Application | Title |
|---|---|
PLoS One PSMD11 modulates circadian clock function through PER and CRY nuclear translocation |