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Halo-Trap Magnetic Particles M-270

Description
Halo-Trap Magnetic Particles M-270 is an affinity resin for immunoprecipitation of Halo-tag fusion proteins. It comprises an anti-Halo-tag Nanobody /VHH conjugated to Magnetic Particles M-270.

Specificity
Halo-tag

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

Product Size Code Price Buy
Product Halo-Trap Magnetic Particles M-270 Size 250 µL (10 reactions) Code otd-10 Price $ 335
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Product Halo-Trap Magnetic Particles M-270 Size 500 µL (20 reactions) Code otd-20 Price $ 550
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Product Halo-Trap Magnetic Particles M-270 Size 2.5 mL (100 reactions) Code otd-100 Price $ 2475
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Product Halo-Trap Magnetic Particles M-270 Size 5 mL (200 reactions) Code otd-200 Price Please inquire
Product Halo-Trap Magnetic Particles M-270 Size 10 mL (400 reactions) Code otd-400 Price Please inquire
Product Halo-Trap Magnetic Particles M-270 Kit Size 500 µL (20 reactions) Code otdk-20 Price $ 715
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Product Halo VHH, recombinant binding protein Size 250 µL Code ot-250 Price $ 365
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Affinity
Dissociation constant KD of 2 nM

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

Specificity
Halo-tag

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

Bead properties
Bead size: ~ 2,8 µm
Storage buffer: 1x PBS, Preservative: 0.09 % sodium azide

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

Compatibility with mass spectrometry
The Halo-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.

How to cite this product
AB_2894835

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 Nano-Traps is very high.

Specifications of Halo-Trap Agarose, Magnetic Agarose, and Magnetic Particles M-270

 

 

Halo-Trap

 

Agarose

Magnetic Agarose

Magnetic Particles M-270

Matrix

Agarose (4% cross-linked)

Magnetic agarose (6% cross linked)

Magnetic Particles M-270

Bead form

Porous

Porous; sold iron core

Solid

Ligand

Halo VHH

Halo VHH

Halo VHH

Halo-tagged protein size*

Small to large size

Small to large size

Small to very large size; no size limitation

Color

White

Black

Brown

Medium particle size

90 µm

40 µm

2.8 µm

Binding capacity

9 µg/ 10 µL

8 µg/ 10 µL

0.5 μg/ 10 μL

Background

Very low

Low

Low

Magnetic separation & automation

No

Yes

Yes

May be centrifuged up to

2,500 x g

800 x g

8,000 x g

* Does depend on protein size and shape, protein multimers, complexes and interaction partners

Benefits of Halo-Trap Magnetic Particles M-270

  • Halo Trap recognizes Halo-fusion proteins that are already bound to chloralkane-based ligands of the Halo-tag, e.g. dyes, biotin, etc.
  • IP and Co-IP of Halo-fusion proteins, even of very large complexes
  • Halo-Trap can be used for affinity purification
  • Acidic elution of bound Halo-fusion proteins
  • Structure and function are characterized

Halo-Trap Magnetic Particles M-270 Kit

The Halo-Trap Magnetic Particles M-270 is also available in a kit, including:

  • Halo-Trap Magnetic Particles M-270
  • lysis*, wash and elution buffers

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

Which Halo-Trap should I use?

•    Halo-Trap Agarose, when lowest background and high binding capacity IP is needed. 
•    Halo-Trap Magnetic Agarose, when magnetic separation and high binding capacity IP is needed.
•    Halo-Trap Magnetic Particles M-270, when very large proteins/complexes are investigated, and magnetic separation is needed for IP.

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