Detecting Histone H3 Citrullination

Antibodies for detecting citrullinated histone H3 (R2 citrullination, Ab176843 and R2-8-17 citrullination, Ab5103), total histone H3 (Ab24834), NCF1, NCF2, and NCF4 were from Abcam (Cambridge, MA). Antibodies for detecting PAD4 (MABE254) and citrullination (F95) were from EMD Millipore (Billerica, MA). Antibodies for PAD IP were developed as previously described^{40 (link)}. Recombinant human PAD2 and recombinant human PAD4 were generated in-house. Recombinant human histone H3, pan PAD inhibitor BB-Cl-amidine, and PAD4 inhibitor GSK484 were from Cayman Chemicals (Ann Arbor, MI). The cell fraction isolation kit was from Cell Signaling (Danvers, MA). Protein G beads, DPI, ionomycin, PMA, Hoechst 33342, HRP-labeled anti-mouse immunoglobulin G (IgG), and anti-rabbit IgG secondary antibodies were from Invitrogen (Carlsbad, CA).

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Zhou Y., An L.L., Chaerkady R., Mittereder N., Clarke L., Cohen T.S., Chen B., Hess S., Sims G.P, & Mustelin T. (2018). Evidence for a direct link between PAD4-mediated citrullination and the oxidative burst in human neutrophils. Scientific Reports, 8, 15228.

Publication 2018

Ab176843 Ab5103 Ab24834 PAD4 inhibitor GSK484 Protein G beads ionomycin PMA Hoechst 33342 anti-rabbit IgG secondary antibodies

Anti antibodies Antibodies Cayman Cell isolation Citrullination Cl amidine Histone h3 Hoechst 33342 Human Immunoglobulin g Ionomycin Mouse Ncf1 Ncf4 Protein g Rabbit

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Variable analysis

independent variables

Recombinant human PAD2
Recombinant human PAD4
Pan PAD inhibitor BB-Cl-amidine
PAD4 inhibitor GSK484

dependent variables

Citrullinated histone H3 (R2 citrullination, Ab176843 and R2-8-17 citrullination, Ab5103)
Total histone H3 (Ab24834)
PAD4 (MABE254)
Citrullination (F95)

control variables

Recombinant human histone H3
Cell fraction isolation kit from Cell Signaling (Danvers, MA)
Protein G beads, DPI, ionomycin, PMA, Hoechst 33342, HRP-labeled anti-mouse immunoglobulin G (IgG), and anti-rabbit IgG secondary antibodies from Invitrogen (Carlsbad, CA)

positive controls

Antibodies for PAD IP developed as previously described

negative controls

Not explicitly mentioned

Annotations

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