TurboGFP-Trap Agarose

Description
TurboGFP-Trap Agarose is an affinity bead for immunoprecipitation (IP) of TurboGFP fusion proteins
It comprises an anti-TurboGFP VHH/ Nanobody coupled to agarose beads.

Specificity
TurboGFP derived from CopGFP from the copepod Pontellina plumata.
The TurboGFP-Trap does not bind to jellyfish GFP and derivatives, see:  Fluorescent protein specificity table

Applications
Immunoprecipitation/ Co-IP
Mass spectrometry
ELISA
On-bead enzyme assays

 

Product Size Code Price Buy
Product TurboGFP-Trap Agarose Size 250 µL (10 reactions) Code tbta-10 Price $ 275
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Product TurboGFP-Trap Agarose Size 500 µL (20 reactions) Code tbta-20 Price $ 475
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Product TurboGFP-Trap Agarose Size 2.5 mL (100 reactions) Code tbta-100 Price $ 2050
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Product TurboGFP-Trap Agarose Size 5 mL (200 reactions) Code tbta-200 Price Please inquire
Product TurboGFP-Trap Agarose Size 10 mL (400 reactions) Code tbta-400 Price Please inquire
Product TurboGFP-Trap Agarose Kit Size 500 µL (20 reactions) Code tbtak-20 Price $ 595
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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 TurboGFP VHH, recombinant binding protein Size 250 µL Code tbt-250 Price $ 275
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Affinity
Dissociation constant KD of 0.5 nM

Wash buffer compatibility
6 M urea, 2 M NaCl, 10 mM DTT, 2 % Nonidet P40 Substitute, 1 % Triton X-100, 0.2 % SDS

Specificity
TurboGFP, CopGFP, Pontellina plumata

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

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

Elution
- SDS sample buffer
- 0.1 mM citrate acid pH 3.0
Instead of elution, we recommend on-bead assays like on-bead digestion for MS analysis.

Compatibility with mass spectrometry
The TurboGFP-Trap is optimized for on-bead digestion. Complete tryptic digest results in 5-6 peptides.

Storage instructions
Shipped at ambient temperature. Upon receipt store at 4°C. Stable for one year.

Does the GFP-Trap bind TurboGFP?

No, the GFP-Trap doesn't bind TurboGFP. TurboGFP is a green fluorescent protein derived from CopGFP of the copepod Pontellina plumata whereas GFP has been originally isolated from jellyfish Aequorea Victoria. Turbo-GFP shares only ~20 % sequence identity with the commonly used GFP variants.

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 can I elute bound proteins from a trap in their native state?

You can elute your fusion protein of interest with 0.2 M glycine pH 2.5 at room temperature. Pipette the beads up and down for 60-120 seconds and repeat this step. Ensure to neutralize your supernatant immediately afterwards by adding 1 M Tris base pH 10.4.

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

  • Low background
  • NO heavy and light antibody chain contamination
  • High binding affinity
  • Harsh washing conditions

TurboGFP-Trap Agarose Kit

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

  • TurboGFP-Trap Agarose
  • Lysis*, wash and elution buffers

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

Spin columns

Binding Control for Agarose Beads

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