Mono-ADP-Ribose antibody | AbD33204
Anti mono-ADP-ribose antibody, clone AbD33204 (TZA019) used to detect ADP-ribosylated peptides by dot blotting.
Image caption:
SpyTag-based modular antibodies enable sensitive and versatile detection of mono-ADPr.
(F) Dot blot analysis of AbD43647, AbD33205, and AbD33204 antibodies blotted at 0.5, 0.25, 0.125 μg and incubated with the indicated biotinylated ADPribosylated peptides. Peptide binding was detected using anti-Streptavidin HRP antibody.
From: Longarini EJ, Dauben H, Locatelli C, Wondisford AR, Smith R, Muench C, Kolvenbach A, Lynskey ML, Pope A, Bonfiglio JJ, Jurado EP, Fajka-Boja R, Colby T, Schuller M, Ahel I, Timinszky G, O'Sullivan RJ, Huet S, Matic I.
Modular antibodies reveal DNA damage-induced mono-ADP-ribosylation as a second wave of PARP1 signaling.
Mol Cell. 2023 May 18;83(10):1743-1760.e11.
doi: 10.1016/j.molcel.2023.03.027.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Anti mono-ADP-ribose antibody, clone AbD33204 (TZA019) used to detect ADP-ribosylated peptides by immunofluorescence.
Image caption:
SpyTag-based modular antibodies enable sensitive and versatile detection of mono-ADPr.
(G) IF analysis of WT U2OS cells, untreated or treated with 2 mM H2O2 for 10 min, PFA fixed and stained with the indicated antibodies.
From: Longarini EJ, Dauben H, Locatelli C, Wondisford AR, Smith R, Muench C, Kolvenbach A, Lynskey ML, Pope A, Bonfiglio JJ, Jurado EP, Fajka-Boja R, Colby T, Schuller M, Ahel I, Timinszky G, O'Sullivan RJ, Huet S, Matic I.
Modular antibodies reveal DNA damage-induced mono-ADP-ribosylation as a second wave of PARP1 signaling.
Mol Cell. 2023 May 18;83(10):1743-1760.e11.
doi: 10.1016/j.molcel.2023.03.027.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Anti mono-ADP-ribose antibody, clone AbD33204 (TZA019) used to detect ADP-ribosylated peptides by immunofluorescence.
Image caption:
SpyTag-based modular antibodies enable sensitive and versatile detection of mono-ADPr.
(B) ELISA analysis of antibody sensitivity. The antibodies AbD33641, AbD33204, or the derived affinity maturated AbD43647 were incubated at the indicated concentrations with immobilized biotinylated peptides (H3S10unmodified or H3S10ADPr, 61 nM). Graphs showing mean ± SEM of n=2 technical replicates. (C) ELISA analysis of antibody specificity. The antibodies AbD33641, AbD33204, or the derived affinity maturated AbD43647 were incubated at 0.2 µg/ml with biotinylated free-poly-ADP-ribose at the indicated concentrations. Graphs showing mean ± SEM of n=2 technical replicates.
From: Longarini EJ, Dauben H, Locatelli C, Wondisford AR, Smith R, Muench C, Kolvenbach A, Lynskey ML, Pope A, Bonfiglio JJ, Jurado EP, Fajka-Boja R, Colby T, Schuller M, Ahel I, Timinszky G, O'Sullivan RJ, Huet S, Matic I.
Modular antibodies reveal DNA damage-induced mono-ADP-ribosylation as a second wave of PARP1 signaling.
Mol Cell. 2023 May 18;83(10):1743-1760.e11.
doi: 10.1016/j.molcel.2023.03.027.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Anti mono-ADP-ribose antibody, clone AbD33204 (TZA019) used to detect ADP-ribosylated peptides by western blotting.
Image caption:
SpyTag-based modular antibodies enable sensitive and versatile detection of mono-ADPr.
(A) Immunoblotting images showing comparison between IgG and HRP-coupled format for the indicated antibodies
From: Longarini EJ, Dauben H, Locatelli C, Wondisford AR, Smith R, Muench C, Kolvenbach A, Lynskey ML, Pope A, Bonfiglio JJ, Jurado EP, Fajka-Boja R, Colby T, Schuller M, Ahel I, Timinszky G, O'Sullivan RJ, Huet S, Matic I.
Modular antibodies reveal DNA damage-induced mono-ADP-ribosylation as a second wave of PARP1 signaling.
Mol Cell. 2023 May 18;83(10):1743-1760.e11.
doi: 10.1016/j.molcel.2023.03.027.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Anti mono-ADP-ribose antibody, clone AbD33204 (TZA019) used to detect ADP-ribosylated peptides by western blotting.
Image caption:
SpyTag-based modular antibodies enable sensitive and versatile detection of mono-ADPr.
(E) Dot blot analysis of in vitro automodified (i) PARP1 WT for poly-ADPr, (ii) PARP1-E988Q (PARP1-EQ) mutant in the absence of HPF1 for mono-ADPr on Asp/Glu, and (iii) PARP1-E988Q mutant in the presence of HPF1 WT for mono-ADPr on Ser. Bottom panels (high-exposed immunoblots of the top panels) show poly-ADPr of PARP1 WT.
From: Longarini EJ, Dauben H, Locatelli C, Wondisford AR, Smith R, Muench C, Kolvenbach A, Lynskey ML, Pope A, Bonfiglio JJ, Jurado EP, Fajka-Boja R, Colby T, Schuller M, Ahel I, Timinszky G, O'Sullivan RJ, Huet S, Matic I.
Modular antibodies reveal DNA damage-induced mono-ADP-ribosylation as a second wave of PARP1 signaling.
Mol Cell. 2023 May 18;83(10):1743-1760.e11.
doi: 10.1016/j.molcel.2023.03.027.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Anti mono-ADP-ribose antibody, clone AbD33204 (TZA019) used to detect ADP-ribosylated peptides in a competition ELISA.
Image caption:
SpyTag-based modular antibodies enable sensitive and versatile detection of mono-ADPr.
(E) Dot blot analysis of in vitro automodified (i) PARP1 WT for poly-ADPr, (ii) PARP1-E988Q (PARP1-EQ) mutant in the absence of HPF1 for mono-ADPr on Asp/Glu, and (iii) PARP1-E988Q mutant in the presence of HPF1 WT for mono-ADPr on Ser. Bottom panels (high-exposed immunoblots of the top panels) show poly-ADPr of PARP1 WT.
From: Longarini EJ, Dauben H, Locatelli C, Wondisford AR, Smith R, Muench C, Kolvenbach A, Lynskey ML, Pope A, Bonfiglio JJ, Jurado EP, Fajka-Boja R, Colby T, Schuller M, Ahel I, Timinszky G, O'Sullivan RJ, Huet S, Matic I.
Modular antibodies reveal DNA damage-induced mono-ADP-ribosylation as a second wave of PARP1 signaling.
Mol Cell. 2023 May 18;83(10):1743-1760.e11.
doi: 10.1016/j.molcel.2023.03.027.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Anti mono-ADP-ribose antibody, clone AbD33204 (TZA019) used to detect ADP-ribosylated peptides by immunofluorescence.
Image caption:
SpyTag-based modular antibodies enable sensitive and versatile detection of mono-ADPr.
(H) IF images of WT, ARH3-KO, and HPF1-KO U2OS cells transfected with FLAG-tagged WT-TRF1-FokI - to induce telomere-localized DNA damage - or DA-TRF1-FokI (D450A, catalytic dead mutant which does not induce DNA damage), fixed with methanol/acetone and stained with the indicated antibodies. Bottom: corresponding quantification of of mono-ADPr positive telomeres (%). Results are mean ± SEM. n = 3 biological replicates, 50 cells. **** P <0.0001 (one-way ANOVA). Scale bars: 5 μm.
From: Longarini EJ, Dauben H, Locatelli C, Wondisford AR, Smith R, Muench C, Kolvenbach A, Lynskey ML, Pope A, Bonfiglio JJ, Jurado EP, Fajka-Boja R, Colby T, Schuller M, Ahel I, Timinszky G, O'Sullivan RJ, Huet S, Matic I.
Modular antibodies reveal DNA damage-induced mono-ADP-ribosylation as a second wave of PARP1 signaling.
Mol Cell. 2023 May 18;83(10):1743-1760.e11.
doi: 10.1016/j.molcel.2023.03.027.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
anti Mono-ADP-Ribose
- Product Type
- Monoclonal Antibody
- Clone
- AbD33204
- Isotype
- Fab antibody
- Specificity
- Mono-ADP-Ribose
Antibody Quality and Performance Guarantee
| Anti mono-ADP-ribose antibody, clone AbD33204, recognizes mono-adenosine diphosphate (ADP)-ribosylation. ADP-ribosylation is a reversible post-translational modification that occurs in multicellular organisms as well as some lower unicellular eukaryotes, but is absent in prokaryotes and yeast (Bürkle 2005). ADP ribosylation has been shown to play critical roles in many physiological and pathological processes, including bacterial pathogenesis and signaling and metabolism to control chromatin-related processes including transcription and DNA repair (Bonfiglio et al. 2020, Bütepage et al. 2015). Members of the ADP-ribosyltransferase (ART) superfamily of proteins including the poly(ADP-ribose) polymerases (PARPs) subfamily, catalyze the transfer of ADP-ribose from nicotinamide adenine dinucleotide (NAD+) onto substrate protein via N-, O-, or S-glycosidic linkages. These transferases are able to transfer a single ADP-ribose residue to their substrate proteins, in a process known as mono-ADP-ribosylation (Bütepage et al. 2015). The anti mono-ADP-ribose antibody, clone AbD33204 is a mono-selective ADP-ribose antibody which was generated using an H3S10ADPr peptide as the antigen. This monovalent Fab format of AbD33204 is tagged with a SpyTag2 at the C-terminus of the Fab heavy chain, enabling the user to couple it to a SpyCatcher reagent for conversion to alternative formats in less than an hour. It has the same specificity for mono-ADPr with a preference for ubiquitin Arg-ADPr and mono-ADPr catalysed by other PARPs than the human/rabbit IgG chimera antibody HCA354. Another mono-ADPr antibody (HCA355, clone AbD33205) also specifically recognizes mono-ADP-ribose, however it has a preference for Ser-mono-ADP-riboses. |
- Target Species
- Protein/peptide tag
- Product Form
- A monovalent human recombinant Fab selected from the HuCAL® phage display library, Expressed in a proprietary E. coli. strain. The antibody is tagged with a DYKDDDDK tag, a SpyTag version 2 (VPTIVMVDAYKRYK) and a His6-tag (HHHHHH) at the C-terminus of the antibody heavy chain. This antibody is supplied as a liquid.
- Preparation
- Recombinant Fab antibody expressed in E. coli and purified by affinity chromatography
- Source
- E.coli
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.0095% MIT
- Carrier Free
- Yes
- Immunogen
- ARTKQTARKS(ADPr)TGGKAC
- Approx. Protein Concentrations
- Total protein concentration 0.5 mg/ml
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
- Acknowledgements
- This product and/or its use is covered by claims of U.S. patents, and/or pending U.S. and non-U.S. patent applications owned by or under license to Bio-Rad Laboratories, Inc. See bio-rad.com/en-us/trademarks for details.
His-tag is a registered trademark of EMD Biosciences
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended. Should this product contain a precipitate we recommend microcentrifugation before use.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended. Should this product contain a precipitate we recommend microcentrifugation before use.
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA |  |
||
| Immunofluorescence | |
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| Western Blotting | |
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
References for Mono-ADP-Ribose antibody
-
Bonfiglio, J.J. et al. (2020) An HPF1/PARP1-Based Chemical Biology Strategy for Exploring ADP-Ribosylation.
Cell. 183 (4): 1086-1102.e23. -
Longarini, E.J. et al. (2023) Modular antibodies reveal DNA damage-induced mono-ADP-ribosylation as a second wave of PARP1 signaling.
Mol Cell. 83 (10): 1743-1760.e11.
TZA019
162594
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