GST-Trap Agarose

Description
GST-Trap is an affinity resin for immunoprecipitation of GST fusion proteins. It consists of a GST Nanobody/ VHH coupled to agarose beads.

Specificity
Glutathione S-Transferase (GST) from Schistosoma japonicum

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

 

Product Size Code Price Buy
Product GST-Trap Agarose Size 250 µL Code sta-10 Price $ 275
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Product GST-Trap Agarose Size 500 µL (20 reactions) Code sta-20 Price $ 475
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Product GST-Trap Agarose Size 2.5 mL (100 reactions) Code sta-100 Price $ 2050
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Product GST-Trap Agarose Size 5 mL (200 reactions) Code sta-200 Price Please inquire
Product GST-Trap Agarose Size 10 mL (400 reactions) Code sta-400 Price Please inquire
Product GST-Trap Agarose Kit Size 500 µL (20 reactions) Code stak-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 GST VHH, recombinant binding protein Size 250 µL Code st-250 Price $ 275
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Affinity
Dissociation constant KD of 1 nM

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

Specificity
Glutathione S-Transferase (GST), Schistosoma japonicum

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

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

Elution
- SDS sample buffer for SDS/PAGE & Western blot
- 0.2 M glycine pH 2.5
Instead of elution, we recommend on-bead assays like on-bead digestion for MS analysis.

Compatibility with mass spectrometry
The GST-Trap is optimized for on-bead digestion. Complete tryptic digest results in 7-8 peptides.

RRID
AB_2631414

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).

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 are the dissociation constants of the Nano-Traps?

Generally heavy chain antibodies do have high affinities to their antigens with dissociation constants in the low nanomolar down to the picomolar range. ChromoTek has determined the following KD values:
GFP-Trap:  1 pM, picomolar (10-12 molar)*
RFP-Trap:  5 nM, nanomolar (10-9 molar)
MBP-Trap:  4 nM, nanomolar (10-9 molar)
GST-Trap:  1 nM, nanomolar (10-9 molar)*
Myc-Trap (with 2x Myc peptide): 0.5 nM, nanomolar (10-9)
Spot-Trap:  6 nM, nanomolar (10-9 molar)
*Kinetic parameter has been measured using the switchSENSE technology using electro-switchable nanolevers to analyze molecular interactions. switchSENSE is a proprietary technology from Dynamic Biosensors (www.dynamic-biosensors.com).

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

  • Pull-down of GST-fusion proteins at low concentrations
  • Strong binding even under harsh washing conditions
  • No heavy & light chains in your downstream application
  • Higher affinity than glutathione cellulose

GST-Trap Agarose Kit

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

  • GST-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

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

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