The ChromoTek V5-Trap® Magnetic Particles M-270 consists of an anti-V5-tag Nanobody (VHH), which is covalently bound to Magnetic Particles M-270. V5-Trap Magnetic Particles M-270 is used to immunoprecipitate V5-tagged fusion proteins from cell extracts of various organisms like mammals, plants, bacteria, yeast, insects etc
V5-tag amino acid sequence GKPIPNPLLGLDST at the N-terminus, C-terminus, or internal site of the fusion protein.
ChIP/ RIP analysis
On-bead enzyme assays
|Product V5-Trap Magnetic Particles M-270||Size 250 µL (10 reactions)||Code v5td-10||Price $ 335||Buy +|
|Product V5-Trap Magnetic Particles M-270||Size 500 µL (20 reactions)||Code v5td-20||Price $ 550||Buy +|
|Product V5-Trap Magnetic Particles M-270||Size 2.5 mL (100 reactions)||Code v5td-100||Price $ 2475||Buy +|
|Product V5-Trap Magnetic Particles M-270||Size 5 mL (200 reactions)||Code v5td-200||Price Please inquire|
|Product V5-Trap Magnetic Particles M-270||Size 10 mL (400 reactions)||Code v5td-400||Price Please inquire|
|Product V5-Trap Magnetic Particles M-270 Kit||Size 500 µL (20 reactions)||Code v5tdk-20||Price $ 715||Buy +|
|Product V5-peptide||Size 1 mg||Code v5p-1||Price $ 85||Buy +|
|Product V5 VHH, recombinant binding protein||Size 250 µL||Code v5t-250||Price Please inquire|
Dissociation constant KD of 40 nM
Wash buffer compatibility
2 % Nonidet P40 Substitute, 2 % Triton X-100, 150 mM NaCl, 10 mM TCEP, 10 mM DTT, 50 mM Urea
V5-tag sequence motif GKPIPNPLLGLDST
10 µL slurry bind ~ 0,25-0,5 µg of recombinant V5-tagged protein ~30 kDa
Coupled Nanobody/ VHH
recombinant, monoclonal anti-V5-tag single domain antibody (sdAb) fragment
Bead size: ~ 2,8 µm
Storage buffer: 1x PBS, Preservative: 0.09 % sodium azide
- V5-peptide. For details see below.
- SDS sample buffer
- 0.2 M glycine pH 2.5
Please consider whether elution is really needed. We recommend on-bead assays like on-bead digestion for MS analysis.
Compatibility with mass spectrometry
The V5-Trap is optimized for on-bead digestion (see protocol).
How to cite this product
Shipped at ambient temperature. Upon receipt store at 4°C. Stable for one year.
Should I use an N-terminal or C-terminal fusion tag?
Both the N-terminal or C-terminal fusion tag work well with the traps.
How can I avoid unspecific protein interactions binding to the trap?
How many mammalian cells are required for an immunoprecipitation reaction?
For one immunoprecipitation reaction, we recommend using ~10^6 - 10^7 mammalian cells. The yield is also dependent on the expression level of your protein of interest and the interaction partners.
How much cell extract should I use for an immunoprecipitation reaction?
For other type of cells than mammalian cells, we recommend using 0.5 - 1.0 mg of cell extract.
Do I need to elute bound proteins from the beads for mass spectrometry analysis?
Do I need to elute my protein of interest from the beads for enzymatic assays?
What is the amount of trap slurry I need for one immunoprecipitation reaction?
25 µL slurry are sufficient for one pull-down reaction as the affinity of the traps is very high.
- No heavy & light antibody chains
- Pull-down of V5-tagged proteins
- Harsh washing conditions
- Elution of native proteins
- One step immunoprecipitation
- Reproducible results
V5-Trap Magnetic Particles M-270
The V5-Trap Magnetic Particles M-270 is also available in a kit, including:
- V5-Trap Magnetic Particles M-270
- Lysis*, dilution and wash buffers
*The lysis buffer provided is suitable for mammalian cells. For other cell types use another suitable lysis buffer.
Which V5-Trap should I use?
- V5-Trap Agarose, when lowest background and high binding capacity IP is needed.
- V5-Trap Magnetic Agarose, when magnetic separation and high binding capacity IP is needed.
- V5-Trap Magnetic Particles M-270, when very large proteins/complexes are investigated, and magnetic separation is needed for IP.
Only for research applications, not for diagnostic or therapeutic use!