V5-Trap Agarose

Description
V5-TrapTM Agarose is an affinity reagent for immunoprecipitation and purification of V5-tagged proteins.
It consists of an anti-V5 Nanobody/ VHH coupled to agarose beads.

Specificity
V5-tag sequence GKPIPNPLLGLDST at the N-terminus, C-terminus, or internal site of the fusion protein.

Applications
Immunoprecipitation/ Co-IP
Protein purification
Mass spectrometry
ChIP/ RIP analysis
On-bead enzyme assays

Product Size Code Price Buy
Product V5-Trap Agarose Size 250 µL (10 reactions) Code v5ta-10 Price $ 275
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Product V5-Trap Agarose Size 500 µL (20 reactions) Code v5ta-20 Price $ 475
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Product V5-Trap Agarose Size 2.5 mL (100 reactions) Code v5ta-100 Price $ 2050
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Product V5-Trap Agarose Size 5 mL (200 reactions) Code v5ta-200 Price Please inquire
Product V5-Trap Agarose Size 10 mL (400 reactions) Code v5ta-400 Price Please inquire
Product V5-Trap Agarose Kit Size 500 µL (20 reactions) Code v5tak-20 Price $ 595
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Product V5-peptide Size 1 mg Code v5p-1 Price $ 85
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Product Binding Control Agarose Beads Size 500 µL (20 reactions) Code bab-20 Price $ 75
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Product Spin Columns Size 10 units Code sct-10 Price $ 35
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Product Spin Columns Size 20 units Code sct-20 Price $ 60
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Product Spin Columns Size 50 units Code sct-50 Price $ 130
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Product V5 VHH, recombinant binding protein Size 250 µL Code v5t-250 Price Please inquire

Affinity
Dissociation constant KD of 40 nM

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

Specificity
V5-tag sequence motif GKPIPNPLLGLDST

Binding capacity
10 µL slurry bind about 6 µg of recombinant V5-tagged protein (~30 kDa)

Coupled Nanobody/ VHH
Recombinant, monoclonal anti-V5-tag single domain antibody (sdAb) fragment

Bead properties
Bead size: ~ 90 µm (cross-linked 4 % agarose beads)
Storage buffer: 20 % EtOH

Elution
- V5-peptide
- SDS sample buffer
- 0.1 M glycine pH 2.0
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. 

How to cite this product
RRID: AB_2868497

Storage instructions
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?

For preclearing of your sample we recommend to use our binding controls (bab-20 or bmab-20).

Please find more information in our Troubleshooting guide

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?

No, you can directly conduct an on-bead digestion after immunoprecipitation. This procedure allows faster and more efficient sample preparation and a potential higher yield.
Please find more information here:

Preomics kit

Protocol on-bead digestion

Do I need to elute my protein of interest from the beads for enzymatic assays?

No, you can directly perform your enzymatic assay on the beads if the active center is not blocked.

Please find more information in our application note "enzymatic activity assay"

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.

Benefits

  • Efficient and fast pulldown of V5-tagged proteins
  • No heavy & light antibody chains in downstream applications
  • Very low background
  • One step immunoprecipitation
  • Easy elution of native proteins

V5-Trap Agarose Kit

The V5-Trap Agarose is also available in a kit, including:

  • V5-Trap Agarose
  • Lysis*, dilution and wash buffers

*The lysis buffer provided is suitable for mammalian cells. For other cell types use another suitable lysis buffer.

Spin columns

  • Easy and convenient handling
  • Rapid washing and clean elution of bound proteins
  • Simplify the pulldown of your protein
  • Spin columns for V5-Trap Agarose

Binding Control for Agarose Beads

  • Control for unspecific binding of proteins, DNA, etc. to beads
  • Pre-clearing of cell lysate
  • Binding control for V5-Trap Agarose

V5-peptide

The V5-peptide may be used for the elution of native and functional V5 fusion proteins bound to V5-Trap or anti-V5 antibodies.

The V5-peptide  has the amino acid sequence GKPIPNPLLGLDST.

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Only for research applications, not for diagnostic or therapeutic use!