The ChromoTek MBP-Trap Agarose are affinity beads for immunoprecipitation of MBP-fusion proteins.
They comprise MBP Nanobodies/ VHHs coupled to agarose beads.
Maltose binding protein (MBP)
On-bead enzyme assays
ChIP/ RIP analysis
|Product MBP-Trap Agarose||Size 250 µL||Code mbta-10||Price $ 275||Buy +|
|Product MBP-Trap Agarose||Size 500 µL (20 reactions)||Code mbta-20||Price $ 475||Buy +|
|Product MBP-Trap Agarose||Size 2.5 mL (100 reactions)||Code mbta-100||Price $ 2050||Buy +|
|Product MBP-Trap Agarose||Size 5 mL (200 reactions)||Code mbta-200||Price Please inquire|
|Product MBP-Trap Agarose||Size 10 mL (400 reactions)||Code mbta-400||Price Please inquire|
|Product MBP-Trap Agarose Kit||Size 500 µL (20 reactions)||Code mbtak-20||Price $ 595||Buy +|
|Product Binding Control Agarose Beads||Size 500 µL (20 reactions)||Code bab-20||Price $ 75||Buy +|
|Product Spin Columns||Size 10 units||Code sct-10||Price $ 35||Buy +|
|Product Spin Columns||Size 20 units||Code sct-20||Price $ 60||Buy +|
|Product Spin Columns||Size 50 units||Code sct-50||Price $ 130||Buy +|
|Product MBP VHH, recombinant binding protein||Size 250 µL||Code mbt-250||Price Please inquire|
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
Maltose binding protein (MBP), E.coli
Coupled Nanobody/ VHH
Recombinant, monoclonal anti-MBP single domain antibody (sdAb) fragment
Bead size: ~ 90 µm (cross-linked 4% agarose beads)
Storage buffer: 20 % EtOH
- 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.
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 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?
Do I need to elute my protein of interest from the beads for enzymatic assays?
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.
- Immunoprecipitation of MBP-fusion proteins at low concentrations
- Strong binding under harsh washing conditions
- Higher affinity than amylose resin
MBP-Trap Agarose Kit
The MBP-Trap Agarose is also available in a kit, including:
- MBP-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.
- Easy and convenient handling
- Rapid washing and clean elution of bound proteins
- Simplify the pulldown of your protein
- Spin Columns for MBP-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 MBP-Trap Agarose
Only for research applications, not for diagnostic or therapeutic use!