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MABE1016

Sigma-Aldrich

Anti-pan-ADP-ribose binding reagent

from Escherichia coli

Synonym(s):

pan-ADP-ribose binding reagent

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160405
NACRES:
NA.41

biological source

Escherichia coli

Quality Level

antibody form

purified antibody

antibody product type

primary antibodies

species reactivity

human, mouse

species reactivity (predicted by homology)

all

technique(s)

dot blot: suitable
immunoprecipitation (IP): suitable
western blot: suitable

shipped in

dry ice

target post-translational modification

unmodified

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.

Specificity

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

Application

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.
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).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
General Post-translation Modification

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).

Target description

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

Physical form

Format: Purified
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.

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.

Other Notes

Concentration: Please refer to lot specific datasheet.

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.

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Karunakaran Kalesh et al.
Scientific reports, 9(1), 6655-6655 (2019-05-02)
ADP-ribosylation is integral to a diverse range of cellular processes such as DNA repair, chromatin regulation and RNA processing. However, proteome-wide investigation of its cellular functions has been limited due to numerous technical challenges including the complexity of the poly(ADP-ribose)
Daniel Roderer et al.
Nature communications, 10(1), 5263-5263 (2019-11-22)
Tc toxins are bacterial protein complexes that inject cytotoxic enzymes into target cells using a syringe-like mechanism. Tc toxins are composed of a membrane translocator and a cocoon that encapsulates a toxic enzyme. The toxic enzyme varies between Tc toxins
Katie Pollock et al.
Methods in molecular biology (Clifton, N.J.), 1608, 445-473 (2017-07-12)
The poly(ADP-ribose)polymerase (PARP) enzyme tankyrase (TNKS/ARTD5, TNKS2/ARTD6) uses its ankyrin repeat clusters (ARCs) to recognize degenerate peptide motifs in a wide range of proteins, thereby recruiting such proteins and their complexes for scaffolding and/or poly(ADP-ribosyl)ation. Here, we provide guidance for
Polly Gravells et al.
DNA repair, 61, 25-36 (2017-11-28)
Upon DNA binding the poly(ADP-ribose) polymerase family of enzymes (PARPs) add multiple ADP-ribose subunits to themselves and other acceptor proteins. Inhibitors of PARPs have become an exciting and real prospect for monotherapy and as sensitizers to ionising radiation (IR). The
Hideyuki Higashi et al.
Journal of proteome research, 18(4), 1607-1622 (2019-03-09)
ADP-ribosylation is a post-translational modification that, until recently, has remained elusive to study at the cellular level. Previously dependent on radioactive tracers to identify ADP-ribosylation targets, several advances in mass spectrometric workflows now permit global identification of ADP-ribosylated substrates. In

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