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Histone h4

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Histone H4 is a core histone protein found in eukaryotic cell nuclei. It plays a crucial role in the organization and structure of chromatin, which is the complex of DNA and histone proteins that make up the chromosomes. Histone H4 is involved in the formation of the nucleosome, the fundamental unit of chromatin, and it contributes to the regulation of gene expression and other DNA-related processes.

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Lab products found in correlation

Gd2 f4, by Thermo Fisher Scientific (3 mentions) Tlr6.127, by Abcam (3 mentions) Elisa kit for von willebrand factor, by Nanjing Jiancheng (3 mentions) Myeloperoxidase mpo detection kit, by Nanjing Jiancheng (3 mentions) Hta125, by Thermo Fisher Scientific (3 mentions) Gapdh, by Santa Cruz Biotechnology (2 mentions) H4k16ac, by Merck Group (2 mentions) α tubulin, by Merck Group (2 mentions) Mini protean tgx precast protein gel, by Bio-Rad (2 mentions) Ac h4 acetyl k5 k8 k12 k16, by Abcam (2 mentions) β actin, by Santa Cruz Biotechnology (2 mentions) Pyrrolidine dithiocarbamate, by Abcam (1 mentions) Isotype igg, by Santa Cruz Biotechnology (1 mentions) Iκbα pser32, by Cell Signaling Technology (1 mentions) C ebp, by Santa Cruz Biotechnology (1 mentions) Mabe1031, by Merck Group (1 mentions) Nmnat2, by Abcam (1 mentions) Acetyl p53 k379, by Cell Signaling Technology (1 mentions) H3k79me2, by Abcam (1 mentions) Usp22, by Abcam (1 mentions)

Spelling variants of this product

Histones (7 citations) Acetylated histone H4 (5 citations) Acetylated histone 4 (Ac‐H4; (3 citations) Rabbit polyclonal anti-acetyl-histone H4, (2 citations) Rabbit anti-acetyl-histone H4 Lys12 (H4K12Ac) IgG (2 citations) Acetyl histone H4 (Lys5/8/12/16) (2 citations) Anti-acetylated histone H4 K16 (2 citations) Anti-acetyl-histone H4 Ab (2 citations) Human histone H4 (2 citations) Anti-histone H4 (citrulline 3) antibody (2 citations)

17 protocols using histone h4

1

Investigating HPSE and NF-κB Signaling Pathways

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The primary antibodies for HPSE (latent precursor: 65 kDa, active HPSE: 50 kDa), GAPDH, and isotype IgG were purchased from Santa Cruz (Dallas, Texas, USA); the primary antibodies for lamin B1, NF-κB p65, IκBα, and IκBα pSer32 from Cell Signaling Technology (Danvers, Massachusetts, USA); a primary antibody for heparan sulfate proteoglycan was purchased from Bioss (Woburn, Massachusetts, USA); blocking antibodies against TLR1 (GD2.F4), TLR2 (TL2.1), and TLR4 (HTA125) from eBioscience (San Diego, CA, USA); a blocking antibody against TLR6 (TLR6.127) from Abcam (Cambridge, UK); a mouse Histone H4 ELISA kit was purchased from USCN Life Science (Wuhan, China); a mouse HPSE ELISA kit was purchased from Biorbyt (Cambridge, UK). A blocking antibody against Histone H4 (anti-H4) was prepared following the previously described protocol [15 (link)]. Histone H4 was purchased from Millipore (Billerica, MA, USA); and the NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) was obtained from Abcam (Waltham, MA, USA). All other reagents were purchased from Sigma (St. Louis, MO, USA), unless stated otherwise.
Zhang Y., Xu F., Guan L., Chen M., Zhao Y., Guo L., Li X., Zheng Y., Gao A, & Li S. (2022). Histone H4 induces heparan sulfate degradation by activating heparanase in chlorine gas-induced acute respiratory distress syndrome. Respiratory Research, 23, 14.
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2

Rabbit Polyclonal Antibody Generation and Characterization

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The custom rabbit polyclonal antiserum against PARP-1 used for Western blotting and ChIP assays was generated by using a purified recombinant antigen comprising the amino-terminal half of PARP-1 (42 (link)) (now available from Active Motif; cat. no. 39559). The custom rabbit polyclonal antiserum against NMNAT-1 was raised against purified recombinant human and mouse NMNAT-1 (Pocono Rabbit Farm and Laboratory). The custom recombinant antibody-like anti-poly-ADP-ribose binding reagent (anti-PAR) was generated and purified in-house (now available from EMD Millipore, MABE1031). The other antibodies used were as follows: C/EBP (Santa Cruz, sc-150X), NMNAT-2 (Abcam, ab56980), -Tubulin (Abcam, ab6046), SIRT1 [custom rabbit polyclonal antiserum raised against mouse SIRT1 (35 (link))], acetyl-p53 K379 (Cell signaling, #2570), p53 (Cell signaling, #2524), H4K16Ac (Millipore, 07–329), Histone H4 (Millipore, 07–108), rabbit IgG (Invitrogen, 10500C), goat anti-rabbit HRP-conjugated IgG (Pierce, 31460), and goat anti-mouse HRP-conjugated IgG (Pierce, 31430).
Ryu K.W., Nandu T., Kim J., Challa S., DeBerardinis R.J, & Kraus W.L. (2018). Metabolic Regulation of Transcription Through Compartmentalized NAD+ Biosynthesis. Science (New York, N.Y.), 360(6389), eaan5780.
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3

Chromatin Immunoprecipitation Protocol

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The antibodies used in this study were specific for USP22 (Abcam), RING1B (MBL international), RING1A (Santa Cruz), BMI1 (Cell signaling technology), α-tubulin (Sigma), GAPDH (Advanced immunochemical inc.), histone H4 (Millipore), RHA (RNA Helicase A) (22 (link)), H3K4me3 (Cell signaling technology), H3K79me2 (Abcam), RNAPII 8WG16 (23 (link)), phospho-H3 (Santa Cruz), Rabbit IgG (GE healthcare) and mouse IgG (GE healthcare).
Arora M., Packard C.Z., Banerjee T, & Parvin J.D. (2015). RING1A and BMI1 bookmark active genes via ubiquitination of chromatin-associated proteins. Nucleic Acids Research, 44(5), 2136-2144.
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4

Western Blot Analysis of EMT Markers

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A549 cells were lysed in PRO-PREPTM protein extraction solution (iNtRON Biotechnology, Seongnam, Korea). Lysates were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred onto polyvinyl difluoride membranes (Millipore Inc., Billerica, MA). Membranes were blocked with a 5% skim milk solution and incubated with the following antibodies: E-cadherin, vimentin, α-SMA, fibronectin, snail, slug (Santa Cruz, CA), HDAC2, HDAC4, ac-histone H3, histone H3, ac-histone H4, histone H4 (Upstate, Millipore Inc.), and β-actin (Santa Cruz, CA). The blots were visualized with HRP-conjugated secondary antibodies and an ECL system (Pierce, Rockford, IL).
Park I.H., Kang J.H., Shin J.M, & Lee H.M. (2016). Trichostatin A Inhibits Epithelial Mesenchymal Transition Induced by TGF-β1 in Airway Epithelium. PLoS ONE, 11(8), e0162058.
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5

Nuclei Isolation and Protein Analysis

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Two grams of 2‐week‐old seedlings were collected, and nuclei were isolated according to the ChIP protocol (Gendrel, Lippman, Martienssen, & Colot, 2005) but without the tissue fixation step. The nuclear protein was released by dissolving the nuclei preparation in 300 μl of lysis buffer (50 mM Tris‐HCl, 10 mM EDTA, 1% SDS, and 1× protease inhibitors) and then sonicated. The protein solution was centrifuged at 16,000 g for 10 min at 4°C to remove debris. Proteins were resolved on a 4%–20% Mini‐PROTEAN TGX Precast Protein Gel (Bio‐Rad) by electrophoresis and detected by antibody to GFP (Abcam, ab290; 1:20,000 dilution) and histone H4 (Millipore, 07‐108; 1:20,000 dilution). histone H4 was used as the loading control.
Shu J., Chen C., Thapa R.K., Bian S., Nguyen V., Yu K., Yuan Z., Liu J., Kohalmi S.E., Li C, & Cui Y. (2019). Genome‐wide occupancy of histone H3K27 methyltransferases CURLY LEAF and SWINGER in Arabidopsis seedlings. Plant Direct, 3(1), e00100.
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6

Isolation and Analysis of Plant Nuclear Proteins

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Two grams of 14-d-old seedlings were collected, and nuclei were isolated according to the ChIP protocol but without the tissue fixation step. Nuclear proteins were released by incubating the nuclei preparation in 120 μl of lysis buffer (50 mM Tris-HCl, 10 mM EDTA, 1% SDS, and 1× protease inhibitors) for 3 h at 4 °C. The extract was then diluted with 1 volume of ChIP dilution buffer (16.7 mM Tris-HCl, 167 mM NaCl, and 1.1% Triton X-100, pH 8.0) and centrifuged at 15,000g for 10 min at 4 °C to remove debris. Proteins were resolved on a 4–20% Mini-PROTEAN TGX Precast Protein Gel (Bio-Rad) by electrophoresis and detected by antibody to GFP (Invitrogen, A11122; 1:5,000 dilution), HA (Sigma, H6908; 1:5,000 dilution), FLAG (Sigma, F7425; 1:5,000 dilution), or histone H4 (Millipore, 07-108; 1:20,000 dilution). histone H4 was used as the loading control. Quantification of protein signal was performed using ImageJ software. Uncropped scans of immunoblotting results are shown in Supplementary Data 6.
Li C., Gu L., Gao L., Chen C., Wei C.Q., Qiu Q., Chien C.W., Wang S., Jiang L., Ai L.F., Chen C.Y., Yang S., Nguyen V., Qi Y., Snyder M.P., Burlingame A.L., Kohalmi S.E., Huang S., Cao X., Wang Z.Y., Wu K., Chen X, & Cui Y. (2016). Concerted genomic targeting of H3K27 demethylase REF6 and chromatin-remodeling ATPase BRM in Arabidopsis. Nature genetics, 48(6), 687-693.
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7

Western Blot Analysis of Fibroblast Cultures

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Fibroblast cultures were harvested at the end of the incubation, and equal amounts of whole cell lysates were subjected to Western analysis, or were first immunoprecipitated with the indicated antibodies followed by Western analysis [4 (link)]. After electrophoresis proteins were then transferred to polyvinylidene difluoride (PVDF) membranes, blocked with 10% fat-free milk in TBST buffer (20 mM Tris HCl, 137 mM NaCl, and 0.05% Tween 20), and incubated with the following primary antibodies: Nrf2, Keap1, Smad1/2/3 (each at 1:200, Santa Cruz Biotechnology, Santa Cruz, CA), p300, histone H4 (1:1000, Millipore), GAPDH (1:3000, Invitrogen), α-SMA (1:2000; Sigma) or Type I collagen (1:400; Southern Biotechnology, Birmingham, AL). Antigen–antibody complexes were visualized by chemiluminescence (Pierce Biotechnology, Rockford, IL), and band intensities were quantified using ImageJ software (http://rsb.info.nih.gov/ij/).
Wei J., Zhu H., Lord G., Bhattachayya M., Jones B.M., Allaway G., Biswal S.S., Korman B., Marangoni R.G., Tourtellotte W.G, & Varga J. (2016). Nrf2 exerts cell-autonomous anti-fibrotic effects: compromised function in systemic sclerosis and therapeutic rescue with a novel heterocyclic chalcone derivative. Translational research : the journal of laboratory and clinical medicine, 183, 71-86.e1.
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8

Histone H4 Regulation of Inflammatory Pathways

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Histone H4 was purchased from Millipore (Billerica, MA, USA). Blocking antibodies against TLR1 (GD2.F4), TLR2 (TL2.1), and TLR4 (HTA125) were purchased from eBioscience (San Diego, CA, USA). Antibodies for TNF-α, IL-1β, P-selectin, Ly6G, H4, Ac-H4(acetyl K5 + K8 + K12 + K16) and blocking antibody against TLR6 (TLR6.127) were purchased from Abcam (Cambridge, UK). An ELISA kit for von Willebrand factor (vWF) and a myeloperoxidase (MPO) detection kit were purchased from Jiancheng Biotech (Nanjing, China). The blocking antibody against Histone H4 (anti-H4) was prepared following the previously described protocol involving autoimmune mice [14 (link)].
Zhang Y., Zhao J., Guan L., Mao L., Li S, & Zhao J. (2020). Histone H4 aggravates inflammatory injury through TLR4 in chlorine gas-induced acute respiratory distress syndrome. Journal of Occupational Medicine and Toxicology (London, England), 15, 31.
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9

Immunoprecipitation and Western Blotting of p49/STRAP

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The expression plasmid constructs containing p49/STRAP and empty vector were transfected into C2C12 cells using Lipofectamine 2000 (Invitrogen) as previously described (Zhang et al., 2004 (link)). At 24 h after the transfection, cells were harvested, and the whole-cell lysate was isolated. The immunoprecipitation and Western blotting were carried out as previously described (Zhang et al., 2004 (link)). Antibodies that were employed included p49/STRAP antibody (Zhang et al., 2004 (link)), Histone H4 (Millipore), anti-acetyl-Histone H4 (Lys16) (Millipore), MFN1 and MFN2 (Santa Cruz Biotechnology) antibodies. Secondary antibodies are from LI-COR Inc. The Western bots were imaged and the bands were analyzed using Odyssey Imaging Systems (LI-COR Inc.).
Zhang X., Williams E.D., Azhar G., Rogers S.C, & Wei J.Y. (2016). Does p49/STRAP, a SRF-binding protein (SRFBP1), Modulate Cardiac Mitochondrial Function in Aging?. Experimental gerontology, 82, 150-159.
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10

Comprehensive Antibody Panel for Drosophila

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We utilised the following primary antibodies directed against Drosophila Mle (in-house), glorund (DSHB 5B7_C), Squid (DSHB 2G9-c), Rump (DSHB 5G4), Histone H3 (Active Motif 39763) and Beta Actin (Santa Cruz (I-19) sc-1616) as well as antibodies recognising human DHX9 (Abcam ab183731), HNRNPM1–4 (Santa Cruz sc-20002), EIF-4A1 (Abcam EPR14506/Ab185946), KHDRBS1 (Sigma S9575), Beta Actin (Santa Cruz (I-19) sc-1616), Histone H3 (Active Motif 39763), histone H4 (Millipore 05–858), OXPHOS Rodent WB Antibody Cocktail (Abcam ab110413- to detect Complex I member NDUF88, Complex II member SDHB, Complex III member UQCRC2, Complex IV member MTCO1 and Complex V member ATP5A1), OXPHOS complex II member SDHA (Invitrogen 459200), TOMM20 (Santa Cruz sc-11415), XRCC5 (Invitrogen MA5–15873), MSH6 (Cell Signalling 5424P), GAPDH (Bethyl A300-641A), HUR (3A2) (Santa Cruz 5261) and FLAG HRP (Sigma A8592). All antibodies were used at a dilution of 1:1000 in 5% fat free milk powder dissolved in 0.3% Tween-20 phosphate buffered saline (Supplementary Figs 19–21).
Panhale A., Richter F.M., Ramírez F., Shvedunova M., Manke T., Mittler G, & Akhtar A. (2019). CAPRI enables comparison of evolutionarily conserved RNA interacting regions. Nature Communications, 10, 2682.
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