Ethenoadenosine Antibody (1G4) - BSA Free

Flow Cytometry: Ethenoadenosine Antibody (1G4) [NB600-442] - Chicken erythrocytes (1% v/v), isolated by density gradient centrifugation, were transiently transfected with a Flag-tagged chicken ART4 containing plasmid or the empty plasmid (pSecTag B), and incubated at 37C in the presence or absence of 200 uM etheno-NAD+ for 30 min. After washing, cells were stained with an etheno-adenosine specific antibody (1G4) or an anti-Flag antibody and the respective isotype control stained with the 1G4 antibody. Image collected and cropped by CiteAb from the following publication (//bmcmolbiol.biomedcentral.com/articles/10.1186/1471-2199-9-86), licensed under a CC-BY license.

Flow Cytometry: Ethenoadenosine Antibody (1G4) [NB600-442] - Detection of ADP-ribosylated proteins by flow cytometry. C-33A cells (1 x 10^7/mL), transiently transfected with a Flag-tagged chicken ART4 containing plasmid, were incubated at 37C in the presence or absence of 200 uM etheno-NAD+ for 30 min. After washing, cells were stained with an etheno-adenosine specific antibody (1G4). Image collected and cropped by CiteAb from the following publication (//bmcmolbiol.biomedcentral.com/articles/10.1186/1471-2199-9-86), licensed under a CC-BY license.

Flow Cytometry: Ethenoadenosine Antibody (1G4) [NB600-442] - Etheno-ADP-ribosylation of cell-surface proteins on C3H-10T1/2 cells. C3H-10T 1/2 cells at different stages of differentiation (progenitors, MyoD-myoblasts, myotubes) were incubated for 30 minutes in the absence (dashed lines) or presence (solid lines) of 100 uM etheno-NAD. Cells were then washed, incubated with the primary monoclonal antibody 1G4, stained with a FITC-conjugated goat-anti-mouse secondary antibody and subjected to FACS analysis. Data show one representative experiments out of three. Image collected and cropped by CiteAb from the following publication (//bmcmolbiol.biomedcentral.com/articles/10.1186/1471-2199-9-91), licensed under a CC-BY license.

• Reactivity: All
• Applications: WB, ELISA, Flow, ICC/IF, IP, CyTOF-ready
• Clone: 1G4
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

Ethenoadenosine Antibody (1G4) - BSA Free Summary

• Immunogen: Ethenoadenosine
• Localization: Nuclear
• Specificity: This is specific for ethenoadenosine and ethenodadenosine.
• Isotype: IgG2 Lambda
• Clonality: Monoclonal
• Host: Mouse
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • ELISA 1:100-1:2000
  • Flow Cytometry 1:10-1:1000. Use reported in scientific literature (PMID 9774627)
  • Immunocytochemistry/ Immunofluorescence 1:50-1:100
  • Immunoprecipitation 1:10-1:500
  • Western Blot 1:800
• Application Notes: This antibody is CyTOF ready.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.1% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for Ethenoadenosine Antibody (1G4) - BSA Free

• C12H13N5O4
• N6-Ethenoadenosine

Background

ADP-ribosylation represents a posttranslational modification that involves transfer of ADP-ribose moiety from NAD+ to a specific amino acid in target protein (greatly modifying protein function) with the release of nicotinamide moiety. It was originally discovered as a mechanism by which potent bacterial toxins, such as Diphtheria toxin and Cholera toxin, modify and inactivate human target proteins, and these toxins were suggested to mimic mammalian endogenous ADP-ribosyltransferases. ART2 is an ecto-ADP-ribosyltransferases family (ART1-ART5) member which distantly resembles ADP-ribosylating bacterial toxins. It is expressed by mature T cells in which it ADP-ribosylates the integrin LFA-1 as well as other cell surface proteins, and this ART2-mediated ADP-ribosylation of T cell surface proteins provides a novel means for triggering T cell apoptosis. Ethenoadenosine moiety is contained in NAD analogue etheno-NAD which serves as an efficient substrate for ADP-ribosyltransferases and ethenoadenosine specific antibody clone 1G4 can be used for detection of ADP-ribosylated amino acids (ADP-ribosylation of cellular proteins) by FACS/FLOW, ICC-IF and Western blot applications.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Ethenoadenosine Antibody (NB600-442)(17)

Publications using NB600-442

• Brown EM, Arellano-Santoyo H, Temple ER Et al. Gut microbiome ADP-ribosyltransferases are widespread phage-encoded fitness factors Cell host & microbe 2021-08-10 [PMID: 34403684]
• Rack JGM, Zorzini V, Zhu Z et al. Viral macrodomains: a structural and evolutionary assessment of the pharmacological potential Open Biol 2020-11-01 [PMID: 33202171] (WB)
• Friedrich M, Bohlig L, Kirschner RD, EngelK, Hauschildt S. Identification of two regulatory binding sites which confer myotube specific expression of the mono-ADP-ribosyltransferase ART1 gene. BMC Mol Biol;9:91. 2008-10-21 [PMID: 18939989] (FLOW)
• Grahnert A, Richter S, Siegert F, Berndt A, Hauschildt S. The orthologue of the acatalytic mammalian ART4 in chicken is an arginine-specific mono-ADP-ribosyltransferase. BMC Mol Biol;9:86. 2008-10-14 [PMID: 18854029] (WB, FLOW, Human, Chicken)
• Hong S, Brass A, Seman M et al. Lipopolysaccharide, IFN-gamma, and IFN-beta induce expression of the thiol-sensitive ART2.1 Ecto-ADP-ribosyltransferase in murine macrophages. J Immunol. 2007-11-01 [PMID: 17947697]
• Koch-Nolte F, Reyelt J, Schossow B et al. Single domain antibodies from llama effectively and specifically block T cell ecto-ADP-ribosyltransferase ART2 2 in vivo. FASEB J. [PMID: 17575259] (ICC/IF, Mouse)
  
  Details:
  Citation using the FITC form of this antibody.
• Liao, SD, Puro, DG. NAD+-induced vasotoxicity in the pericyte-containing microvasculature of the rat retina: effect of diabetes. Invest Ophthalmol Vis Sci;47(11):5032-8. 2006-11-01 [PMID: 17065524] (ICC/IF, Rat)
• Kawamura H, Aswad F, Minagawa M, Govindarajan S, Dennert G. P2X7 receptors regulate NKT cells in autoimmune hepatitis. J Immunol;176(4):2152-60. 2006-02-15 [PMID: 16455971]
• Aswad F, Kawamura H, Dennert G. High sensitivity of CD4+CD25+ regulatory T cells to extracellular metabolites nicotinamide adenine dinucleotide and ATP: a role for P2X7 receptors. J Immunol;175(5):3075-83. 2005-09-01 [PMID: 16116196] (FLOW)
• Kawamura H, Aswad F, Minagawa M, Malone K, Kaslow H, Koch-Nolte F, Schott WH, Leiter EH, Dennert G. P2X7 receptor-dependent and -independent T cell death is induced by nicotinamide adenine dinucleotide. J Immunol;174(4):1971-9. 2005-02-15 [PMID: 15699125] (FLOW, WB, Mouse)
• Krebs, C et al. Flow cytometric and immunoblot assays for cell surface ADP-ribosylation using a monoclonal antibody specific for ethenoadenosine. Anal Biochem;314(1):108-15. 2003-03-01 [PMID: 12633608] (FLOW, WB, ICC/IF, Mouse, Human)
• Ohlrogge, W et al. Generation and characterization of ecto-ADP-ribosyltransferase ART2.1/ART2.2-deficient mice. Mol Cell Biol. 22(21):7535-42. 2002-11-01 [PMID: 12370300]
• Grahnert, A et al. Mono-ADP-ribosyltransferases in human monocytes: regulation by lipopolysaccharide. Biochem J;362(Pt 3):717-23. 2002-03-15 [PMID: 11879200] (ICC/IF, FLOW, Human)
• Ablamunits V, Bridgett M, Duffy T, Haag F, Nissen M, Koch-Nolte F, Leiter H. Changing patterns of cell surface mono (ADP-ribosyl) transferase antigen ART2.2 on resting versus cytopathically-activated T cells in NOD/Lt mice. Diabetologia;44(7):848-58. 2001-07-01 [PMID: 11508269] (FLOW, Mouse)
• Kahl S, Nissen M, Girisch R, Duffy T, Leiter EH, Haag F, Koch-Nolte F. Metalloprotease-mediated shedding of enzymatically active mouse ecto-ADP-ribosyltransferase ART2.2 upon T cell activation. J Immunol;165(8):4463-9. 2000-10-15 [PMID: 11035085] (FLOW, ICC/IF, Mouse)
• Davis RE, Mysore V, Browning JC, Hsieh JC, Lu QA, Katsikis PD. In situ staining for poly(ADP-ribose) polymerase activity using an NAD analogue. J Histochem Cytochem;46(11):1279-89. 1998-11-01 [PMID: 9774627] (ICC/IF, FLOW, Human)
• Young, TL, Santella, RM. Development of techniques to monitor for exposure to vinyl chloride: monoclonal antibodies to ethenoadenosine and ethenocytidine. Carcinogenesis;9(4):589-92. 1988-04-01 [PMID: 3356066]

Review for Ethenoadenosine Antibody (NB600-442) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Yao Wang Flow Mouse 08/03/2021

Summary

• Application: Flow Cytometry
• Sample Tested: Mouse spleenocytes
• Species: Mouse
• Lot: A-3-042721