Description The ChromoTek MBP-Trap Agarose are affinity beads for immunoprecipitation of MBP-fusion proteins. They comprise MBP Nanobodies/ VHHs coupled to agarose beads.

Specificity Maltose binding protein (MBP)

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

Affinity Dissociation constant KD of 4 nM

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

Specificity Maltose binding protein (MBP), E.coli

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

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

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

RRID AB_2631382

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

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

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.

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.

For other type of cells than mammalian cells, we recommend using 0.5 - 1.0 mg of cell extract.

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

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

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).

25 µL slurry are sufficient for one pull-down reaction as the affinity of the traps is very high.

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

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