PARP1-Trap Agarose

Immunoprecipitation of PARP1 with PARP1-Trap.
I: Input, FT: Flow-through, B: Bound

Description
PARP1-Trap Agarose is an affinity reagent for immunoprecipitation of human PARP1.
It consists of a PARP1 Nanobody/ V_HH, which is coupled to agarose beads.

Specificity
Human PARP1, does NOT bind to PARP2, 3, and 9

Applications
Immunoprecipitation (IP) / Co-IP
Mass spectrometry
On-bead enzyme assays

Product	Size	Code	Price	Buy
Product PARP1-Trap Agarose	Size 250 µL (10 reactions)	Code xta-10	Price $ 275	Buy +
Product PARP1-Trap Agarose	Size 500 µL (20 reactions)	Code xta-20	Price $ 475	Buy +
Product PARP1-Trap Agarose	Size 2.5 mL (100 reactions)	Code xta-100	Price $ 2050	Buy +
Product PARP1-Trap Agarose	Size 5 mL (200 reactions)	Code xta-200	Price Please inquire
Product PARP1-Trap Agarose	Size 10 mL (400 reactions)	Code xta-400	Price Please inquire
Product PARP1-Trap Agarose Kit	Size 500 µL (20 reactions)	Code xtak-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 PARP1-Binding-Protein	Size 250 µL	Code xt-250	Price Please inquire

Specificity
Human PARP1
Epitope: Within DNA binding domain of PARP1
Does NOT bind to PARP 2, 3, and 9

Coupled Nanobody/ VHH
Recombinant, monoclonal anti-PARP1 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

Protocol

SDS

PARP1-Trap Agarose SDS (PDF)
PARP1-Trap Agarose Kit SDS (PDF)

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