p53-N-term-Trap Agarose

Description
p53-N-term-Trap Agarose is an affinity reagent for immunoprecipitation of p53.
It consists of a p53 Nanobody/ VHH coupled to agarose beads, and which binds to the N-terminus of p53.

Specificity
p53
Species-Reactivity: Tested on human
Epitope: 1-81 aa

Applications
Immunoprecipitation (IP) / Co-IP
Mass spectrometry

Product Size Code Price Buy
Product p53-N-term-Trap Agarose Size 250 µL (10 reactions) Code pta-10 Price $ 275
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Product p53-N-term-Trap Agarose Size 500 µL (20 reactions) Code pta-20 Price $ 475
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Product p53-N-term-Trap Agarose Size 2.5 mL (100 reactions) Code pta-100 Price $ 2050
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Product p53-N-term-Trap Agarose Size 5 mL (200 reactions) Code pta-200 Price Please inquire
Product p53-N-term-Trap Agarose Size 10 mL (400 reactions) Code pta-400 Price Please inquire
Product p53-N-term-Trap Agarose Kit Size 500 µL (20 reactions) Code ptak-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 p53-N-term-Binding-Protein Size 250 µL Code pt-250 Price $ 275
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Specificity
p53
Species-Reactivity: tested on human
Epitope: aa 1-81

Coupled Nanobody/ VHH
Recombinant, monoclonal anti-p53 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.

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

 

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.

p53-N-term-Trap Agarose Kit

The p53-N-term-Trap Agarose is also available in a kit, including:

  • p53-N-term-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 p53-N-term-Trap Agarose

Binding Control for Agarose Beads

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