Application

Research Sub CategoryGeneral Post-translation Modification

Dot Blot Specificity Analysis: This reagent detected mono(ADPR) on recombinant PARP3 protein, as well as mono(ADPR) and poly(ADPR) on recombinant PARP1 recombinant protein (Lee Kraus, University of Texas Southwestern Medical Center).

Immunoprecipitation Analysis: A representative lot of Anti-pan-ADP-ribose binding reagent immunoprecipitated ADP-ribosylated proteins from nuclear extract (Lee Kraus, University of Texas Southwestern).

Western Blotting Analysis: A representative lot detected auto-ADP-ribosylation activity of PARP1/2/3 and mutants in the presence of NAD+ or various NAD+ analogs (Gibson, B.A., et al. (2016). Science. 353(6294):45-50).

Western Blotting Analysis: A representative lot detected PARP1-catalyzed NELF-E ADP-ribosylation in cell-free enzymatic reactions as well as ADP-ribosylation of exogenously expressed FLAG-tagged NELF-E in HEK293T cells. PARP inhibitor PJ34 or P-TEFb/CDK9 inhibitor flavopiridol treatment decreased cellular NELF-E ADP-ribosylation level (Gibson, B.A., et al. (2016). Science. 353(6294):45-50).

Anti-poly-ADP-ribose binding reagent is a reagent that selectively binds to both mono- and poly- ADP ribose for use in Western Blotting, Immunocytochemistry and Dot Blot.

Research CategoryEpigenetics & Nuclear Function

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Cat. No.MABE1016, Anti-pan-ADP-ribose binding reagent, is a His-tagged recombinant protein fused to rabbit Fc tag, expressed in and purified from Rosetta™(DE3)pLysS strain of E. coli (Cat. No. 70956). Anti-pan-ADP-ribose binding reagent is useful for the affinity detection of both mono- and poly-ADP-ribosylated proteins on membranes in a manner similar to antibody-based Western and dot blot analysis, The rabbit Fc tag allows visualization of the binding with conjugated anti-rabbit secondary antibodies. The Fc tag also allows Anti-pan-ADP-ribose binding reagent to be captured on Protein A resins for affinity pull-down applications.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Ni-NTA agarose

Purified from E. coli by Ni-NTA agarose. Supplied in buffer containing 10 mM Tris pH 7.5, 0.2 M NaCl, 10% Glycerol, 10 mM Imidazole, 1 mM PMSF, 1 mM β-Mercaptoethanol, 10% glycerol without preservatives.

Format: Purified

Quality

Evaluated by Western Blotting on ADP-ribosylated PARP1 and PARP3 recombinant proteins.

Western Blotting Analysis: This reagent detected mono(ADPR) on recombinant PARP3 protein, as well as mono(ADPR) and poly(ADPR) on recombinant PARP1 protein (Lee Kraus, University of Texas Southwestern Medical Center).

Specificity

poly(ADP-ribose) and mono(ADP-ribose)

Storage and Stability

Stable for 1 year at -80°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -80°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

Variable depending on the target proteins and the extend of ADP-ribosylation.