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07-596 Anti-Histone H4 (citrulline 3) Antibody

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      Key Specifications Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      HChIP, ICC, IH(P), WBRbAffinity PurifiedPolyclonal Antibody
      Catalogue Number07-596
      Brand Family Upstate
      Trade Name
      • Upstate
      DescriptionAnti-Histone H4 (citrulline 3) Antibody
      Alternate Names
      • Cit-H4R3
      • H4Cit3
      • Histone H4, Arg3 citrullinated
      Background InformationHistone H4 (UniProt P62805; also known as H4 histone family member A, Histone 1 H4a) is encoded by the HIST1H4A (also known as H4/A, H4FA) gene (Gene ID: 8359) in human. Histone H4 is one of the main histone proteins that contributes to the chromatin structure in both prokaryotic and eukaryotic cells. Histone H4 can undergo several different types of epigenetic modifications that influence cellular processes, including methylation and acetylation on lysine and arginine residues, phosphorylation on serine and threonine residues, as well as conversion Arg3 to a citrulline residue by the peptidylarginine deiminase PAD4-catalyzed deimination reaction (citrullination). Histone H4 Arg3 citrullination (H4Cit3) weakens histone-DNA interaction as a result of a conformational change and a reduced positive charge at the histone H4 N-terminal end. Bacterial infection is known to stimulate H4Cit3 in neutrophils, forming highly decondensed extracellular chromatin structure NET (neutrophil extracellular trap) that could capture and kill microorganisms. On the other hand, citrullinated proteins are recognized as non-self proteins and can activate the immune system. A functional haplotype of the PADI4 gene has been shown to be associated with the pathogenesis of rheumatoid arthritis through induction of the antigenicity of proteins. H4Cit3 is induced in response to DNA damage through the p53-PADI4 pathway, resulting in upregulated H4Cit3 immunoreactivity localized around fragmented nuclei in apoptotic cells. In addition, H4Cit3 level is negatively correlated with p53 protein expression and tumor size in non-small cell lung cancer (NSCLC) tissues, indicating a crucial role of H4Cit3 in carcinogenesis.
      Product Information
      FormatAffinity Purified
      PresentationPurified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
      Quality LevelMQ100
      ApplicationAnti-Histone H4 (Citrulline 3), Cat. No. 07-596, is rabbit polyclonal antibody that detects Histone H4 citrullinated on Arginine 3 and is tested in ChIP, Immunocytochemistry, Immunohistochemistry (Paraffin), Peptide Inhibition Assay & Western Blotting.
      Key Applications
      • Chromatin Immunoprecipitation (ChIP)
      • Immunocytochemistry
      • Immunohistochemistry (Paraffin)
      • Western Blotting
      Application NotesTested Applications

      Immunocytochemistry Analysis (ICC): A 1:500 dilution from a representative lot detected enhanced histone H4 Arg3 citrullination in ionomycin-stimulated primary human neutrophils. (Courtesy of Kerri Mowen, Ph.D., Padlock Therapeutics).

      Immunohistochemistry Analysis (IHC): A 1:250 dilution from a representative lot detected enhanced histone H4 Arg3 citrullination (H4Cit3) in 4% paraformaldehyde-fixed, decalcified, paraffin-embedded hind paw sections from wild-type, but not Pad4-knockout mice 5 days post immunization with arthritogenic serum from K/BxN mice (Courtesy of Kerri Mowen, Ph.D., Padlock Therapeutics).

      Peptide Inhibition Assay: Target band detection in acid extract of DMSO differentiated HL60 cells treated with calcium ionophore A23187 was prevented by preblocking of a representative lot with Histone H4 (citrulline 3) peptide.

      Chromatin Immunoprecipitation (ChIP) Analysis (ChIP): 1 µg from a representative lot immunoprecipitated Histone H4 (Citrulline 3) in chromatin from MCF-7 cells.

      Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
      Biological Information
      ImmunogenKLH-conjugated linear peptide corresponding to the first 10 amino acids of human histone H4 (Met1 removed), where Arginine 3 is citrullinated. C-terminal cysteine was added for conjugation purposes.
      ConcentrationPlease refer to lot specific datasheet.
      SpecificityThis rabbit polyclonal antibody specifically detects Histone H4 citrullinated on Arginine 3.
      Species Reactivity
      • Human
      Species Reactivity NoteHuman. Predicted to react with Mouse based on 100% sequence homology.
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Gene Symbol
      • HIST1H4A
      • H4/A
      • ;H4FA
      • HIST1H4B
      • H4/I
      • ;H4FI
      • HIST1H4C
      • H4/G
      • H4FG
      • HIST1H4D
      • H4/B
      • H4FB
      • HIST1H4E
      • H4/J
      • H4FJ
      • HIST1H4F
      • H4/C
      • H4FC
      • HIST1H4H
      • H4/H
      • H4FH
      • HIST1H4I
      • H4/M
      • H4FM
      • HIST1H4J
      • H4/E
      • H4FE
      • HIST1H4K
      • H4/D
      • H4FD
      • HIST1H4L
      • H4/K
      • H4FK
      • HIST2H4A
      • H4/N
      • H4F2
      • H4FN
      • HIST2H4
      • HIST2H4B
      • H4/O
      • H4FO
      • HIST4H4
      • HIST1H4A
      • H4/A
      • Citrullination
      Purification MethodAffinity purified.
      UniProt Number
      Molecular Weight~12 kDa observed; 11.37 kDa calculated. Uncharacterized bands may be observed in some lysate(s).
      Physicochemical Information
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceEvaluated by Western Blotting in lysate from HL60 cells induced by retinoic acid and treated with calcium Ionophore (A23187).

      Western Blotting Analysis (WB): A 1:2,000 dilution of this antibody detected Histone H4 citrullinated on arginine 3 in lysates from HL60 cells induced by retinoic acid and either untreated (Lane 1) or treated with calcium ionophore (A23187) (Lane 2).
      Usage Statement
      • 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.
      Storage and Shipping Information
      Storage ConditionsRecommend storage at +2°C to +8°C. For long term storage antibodies can be kept at -20°C. Avoid repeated freeze-thaws.
      Packaging Information
      Material Size200 µL
      Transport Information
      Supplemental Information


      Anti-Histone H4 (citrulline 3) Antibody SDS


      Safety Data Sheet (SDS) 

      Anti-Histone H4 (citrulline 3) Antibody Certificates of Analysis

      TitleLot Number
      Anti-Histone H4 (Citrulline 3) 2474906
      Anti-Histone H4 (Citrulline 3) (rabbit immunoaffinity purified IgG) - 2121995 2121995
      Anti-Histone H4 (Citrulline 3) (rabbit immunoaffinity purified IgG) - 2370138 2370138
      Anti-Histone H4 (Citrulline 3) (rabbit immunoaffinity purified IgG) - 2420472 2420472
      Anti-Histone H4 (Citrulline 3) (rabbit immunoaffinity purified IgG) - 2430493 2430493
      Anti-Histone H4 (Citrulline 3) (rabbit immunoaffinity purified IgG) - 2455608 2455608
      Anti-Histone H4 (Citrulline 3) (rabbit immunoaffinity purified IgG) - 2193166 2193166
      Anti-Histone H4 (Citrulline 3) (rabbit immunoaffinity purified IgG) - 2266824 2266824
      Anti-Histone H4 (Citrulline 3) (rabbit immunoaffinity purified IgG) - 2342877 2342877
      Anti-Histone H4 (Citrulline 3) (rabbit immunoaffinity purified IgG) - 3437326 3437326


      Reference overviewApplicationPub Med ID
      Deimination of linker histones links neutrophil extracellular trap release with autoantibodies in systemic autoimmunity.
      Dwivedi, N; Neeli, I; Schall, N; Wan, H; Desiderio, DM; Csernok, E; Thompson, PR; Dali, H; Briand, JP; Muller, S; Radic, M
      FASEB journal : official publication of the Federation of American Societies for Experimental Biology  28  2840-51  2014

      Show Abstract
      24671707 24671707
      PAD4-mediated neutrophil extracellular trap formation is not required for immunity against influenza infection.
      Hemmers, S; Teijaro, JR; Arandjelovic, S; Mowen, KA
      PloS one  6  e22043  2011

      Show Abstract Full Text Article
      21779371 21779371
      Potential role for peptidylarginine deiminase 2 (PAD2) in citrullination of canine mammary epithelial cell histones.
      Cherrington, BD; Morency, E; Struble, AM; Coonrod, SA; Wakshlag, JJ
      PloS one  5  e11768  2010

      Show Abstract Full Text Article
      Western Blotting20668670 20668670
      Analysis of histones in Xenopus laevis. I. A distinct index of enriched variants and modifications exists in each cell type and is remodeled during developmental transitions.
      Shechter, D; Nicklay, JJ; Chitta, RK; Shabanowitz, J; Hunt, DF; Allis, CD
      The Journal of biological chemistry  284  1064-74  2009

      Show Abstract
      18957438 18957438
      Functional connection between deimination and deacetylation of histones.
      Denis, H; Deplus, R; Putmans, P; Yamada, M; Métivier, R; Fuks, F
      Molecular and cellular biology  29  4982-93  2009

      Show Abstract
      19581286 19581286
      Human PAD4 regulates histone arginine methylation levels via demethylimination
      Wang, Y., et al
      Science, 306:279-83 (2004)  2004

      Immunocytochemistry15345777 15345777

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