Myc-Trap Magnetic Agarose

Description
The ChromoTek Myc-Trap® Magnetic Agarose is an affinity resin for immunoprecipitation of Myc-fusion proteins.
It consists of a Myc Nanobody/VHH coupled to magnetic agarose beads.

Specificity
Myc-tag sequence motif EQKLISEEDL at the N-terminus, C-terminus, or internal site of the fusion protein.
Endogenous c-myc is NOT bound.

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

Product Size Code Price Buy
Product Myc-Trap Magnetic Agarose Size 250 µL (10 reactions) Code ytma-10 Price $ 275
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Product Myc-Trap Magnetic Agarose Size 500 µL (20 reactions) Code ytma-20 Price $ 475
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Product Myc-Trap Magnetic Agarose Size 2.5 mL (100 reactions) Code ytma-100 Price $ 2050
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Product Myc-Trap Magnetic Agarose Size 5 mL (200 reactions) Code ytma-200 Price Please inquire
Product Myc-Trap Magnetic Agarose Size 10 mL (400 reactions) Code ytma-400 Price Please inquire
Product Myc-Trap Magnetic Agarose Kit Size 500 µL (20 reactions) Code ytmak-20 Price $ 595
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Product Myc Peptide Size 1 mg Code yp-1 Price $ 85
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Product Myc Peptide Size 10 mg Code yp-10 Price $ 640
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Product 2x Myc Peptide Size 1 mg Code 2yp-1 Price $ 105
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Product Binding Control Magnetic Agarose Beads Size 500 µL (20 reactions) Code bmab-20 Price $ 75
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Product Myc VHH, recombinant binding protein Size 250 µL Code yt-250 Price $ 275
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Affinity
1x Myc-tag: Dissociation constant KD of 500 nM
2x Myc-tag: Dissociation constant KD of 0.5 nM

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

Specificity
Myc-tag sequence motif EQKLISEEDL

Binding capacity
10 µL slurry bind ~ 7 µg of recombinant Myc-tagged MBP

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

Bead properties
Bead size: ~ 40 µm
Storage buffer: 20% EtOH

Elution
- 1x Myc peptide, 2x Myc peptide. For details see below.
- SDS sample buffer
- 0.2 M glycine pH 2.5
Please consider whether elution is really needed. We recommend on-bead assays like on-bead digestion for MS analysis.

Compatibility with mass spectrometry
The Myc-Trap is optimized for on-bead digestion. Complete tryptic digest results in 6-7 peptides.

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

Does the Myc-Trap bind endogenous c-Myc protein?

We could not detect binding of endogenous c-Myc protein to the Myc-Trap. Some epitope residues that have shown to be crucial for binding to the Myc-Trap are buried in the three-dimensional structure of the c-Myc protein. Hence, under native conditions, c-Myc protein is not a suitable binding partner for the Myc-Trap.

How can I gently elute native Myc-tagged protein from the Myc-Trap in its native state?

You can elute your Myc-fusion competitively with 1x or 2x Myc-peptide. Alternatively, you can use 8 M Urea or 0.2 M glycine pH 2.5 at room temperature.

Should I use 1x Myc- or 2x Myc-peptide for elution of the Myc-Trap?

It is likely that the Myc-Trap binds several motifs of a double Myc-tag. Hence, elution of a Myc-tagged fusion protein is more efficient with the 2x Myc-peptide. In addition, the term "Myc-tag" can refer to a single or double Myc-tag. To be on the safe side in this case as well, we would recommend to elute with the 2x Myc-peptide.

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

  • No heavy & light antibody chains
  • Pull-down of Myc-tagged proteins
  • Elution of native proteins
  • One step immunoprecipitation

1x Myc-Tag or 2x Myc-Tag: Binding and Elution

Both, 1x Myc- and 2x Myc-tagged Protein of Interest (POI) can be used for all kind of IPs, Co-IPs, and affinity purification. To further optimize your experiment:

  • Use 1x Myc-tagged POI if you like to effectively or gently elute your protein of interest from Myc-Trap in native conditions.
  • Use 2x Myc-tagged POI if you like to effectively pull-down/bind low expressed/abundant proteins. For downstream assays we recommend on-bead applications like on-bead digestion and on-bead assays.

 

Elution efficiency

  1x Myc-Tag 2x and 3x Myc-Tag
1x Myc-peptide (40 µM) ++ o
2x Myc-peptide (40 µM) +++ +
Urea (8 M) +++ +
Glycine (0.2 M) pH 2.5 + +

Elution of 1x, 2x and 3x Myc-tagged protein from Myc-Trap.

 

Myc-Trap Consideration Aspects

1x Myc-fusion protein

2x and 3x Myc-fusion protein

 
  • Effective one step elution: both 2x Myc-peptide and urea result in complete elution of bound Myc-tagged protein
  • Small elution volume to concentrate Myc-fusion protein of interest
  • Both 1x Myc and 2x Myc-peptides enable gentle elution for recovery of native proteins
  • Fast binding for short incubation time
  • Fast dissociation requires quick wash processes
 
 
  • Effective binding, optimal for low or endogenous expressed 2x Myc-fusion protein
  • Stringent wash conditions can be applied
  • Complete pull-down/ binding
  • Fast binding enables short incubation of 2x Myc-fusion protein
  • Slow dissociation of 2x Myc-fusion protein enables long wash steps
  • Incomplete elution favors on-bead downstream assays & digestion of 2x Myc-fusion protein
 


For details, see white paper

Myc-Trap Magnetic Agarose Kit

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

  • Myc-Trap Magnetic Agarose
  • Lysis*, wash and elution buffers

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

Binding Control for Magnetic Agarose Beads

Myc Peptide

The Myc peptides may be used to elute native and functional Myc-tagged fusion proteins bound to Myc-Trap® or anti-Myc antibodies.

Myc peptide (1x), amino acid sequence: EQKLISEEDL, and Myc peptide (2x), amino acid sequence: EQKLISEEDLEQKLISEEDL.

  • Easy elution of Myc-tagged fusion proteins

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