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Anti h4

Manufactured by Cell Signaling Technology
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Anti-H4 is a laboratory reagent that detects the histone H4 protein. It is a specific antibody that can be used to identify and study the presence and distribution of histone H4 in biological samples.

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Lab products found in correlation

Anti h3, by Cell Signaling Technology (2 mentions) Anti brd4, by Cell Signaling Technology (2 mentions) Anti γh2ax, by Cell Signaling Technology (2 mentions) Anti acetylated lysine, by Cell Signaling Technology (1 mentions) Mouse anti gapdh mab, by Merck Group (1 mentions) 0.22 m nitrocellulose membrane, by Bio-Rad (1 mentions) Anti ha, by Cell Signaling Technology (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Clarity western ecl substrate, by Bio-Rad (1 mentions) Anti mouse hrp conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions) Anti p stat1, by Cell Signaling Technology (1 mentions) Anti p65, by Cell Signaling Technology (1 mentions) Anti p21 10355 1 ap, by Proteintech (1 mentions) Anti p p65, by Cell Signaling Technology (1 mentions) Anti tbk1, by Cell Signaling Technology (1 mentions) Anti p tbk1, by Cell Signaling Technology (1 mentions) Anti h2ax, by Cell Signaling Technology (1 mentions) Anti sting, by Cell Signaling Technology (1 mentions) Anti stat1, by Cell Signaling Technology (1 mentions) Anti tom20, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

Anti-Histone H4 (11 citations) Anti-acetyl histone H4 (6 citations) Anti-Acetyl-Histone H4(Lys12) (2 citations) Anti-Histone H4 antibody (2 citations)

5 protocols using anti h4

1

Antibodies for Western Blotting Analysis

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The following antibodies (Abs) were used for Western blotting: rabbit anti-pan-Akt mAb (#4691), anti-p-Akt (T308) mAb (#4056), anti-p-Akt (S473) mAb (#4060), anti-pan-ERK mAb (#4695), anti-p-ERK (T202/Y204) mAb (#4376), anti-BRD4 (#13,440), anti-H3(#4499), anti-H4(#13,919), anti-acetyl-H3(Lys9/Lys14) (#9677), anti-acetyl-H3 (Lys27) (#8173), anti-acetyl-H4 (Lys5) (#8647), anti-acetyl-H4 (Lys8) (#2594), anti-acetyl-H4 (Lys16) (#13,534) and anti-acetylated-lysine (#9441); all of these were purchased from Cell Signaling Technology (Danvers, MA, USA). Mouse anti-GAPDH mAb (G8795) was purchased from Sigma-Aldrich. Goat anti-PD-L1 (AF1019) was purchased from R&D Systems (Minneapolis, MN, USA).
Shi Y., Fu Y., Zhang X., Zhao G., Yao Y., Guo Y., Ma G., Bai S, & Li H. (2020). Romidepsin (FK228) regulates the expression of the immune checkpoint ligand PD-L1 and suppresses cellular immune functions in colon cancer. Cancer Immunology, Immunotherapy, 70(1), 61-73.
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2

Western Blot Analysis of HA and H4 Proteins

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C33-EV, C33-E616, and C33-E716 cells were cultured in 60 mm dishes and after 24 h lysed using 300 µL of RIPA buffer (100 mM Tris pH 8.0, 50 mM NaCl2, 0.5% Nonidet P-40, and protease inhibitor cocktail (Roche, Basel, CH)). A total of 20 μg of cell protein extracts were analyzed by SDS-PAGE gels (10–12%) and blotted onto a 0.22 µm nitrocellulose membrane (Bio-Rad, Hercules, CA, USA). Membranes were blocked with 10% skimmed milk in TBS-0.1% Tween 20 for 1 h at room temperature, followed by incubation with anti-HA (Cell Signaling, Danvers, MA, USA) and anti-H4 (Cell Signaling, Danvers, MA, USA) primary antibodies diluted 1:1000 and 1:20,000, respectively. After washing three times with TBS-0.1% Tween 20, membranes were incubated with HRP-conjugated secondary anti-mouse antibody (Santa Cruz, Bio., Dallas, TX, USA) in a dilution 1:10,000. Proteins were visualized utilizing the Clarity™ Western ECL Substrate (Bio-Rad, Hercules, CA, USA), according to the manufacturer’s instructions. Then, membranes were visualized and analyzed in the iBright FL1500 imagining system (Invitrogen, Waltham, MA, USA).
Olmedo-Nieva L., Muñoz-Bello J.O., Martínez-Ramírez I., Martínez-Gutiérrez A.D., Ortiz-Pedraza Y., González-Espinosa C., Madrid-Marina V., Torres-Poveda K., Bahena-Roman M, & Lizano M. (2022). RIPOR2 Expression Decreased by HPV-16 E6 and E7 Oncoproteins: An Opportunity in the Search for Prognostic Biomarkers in Cervical Cancer. Cells, 11(23), 3942.
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3

Antibodies for Chromatin and Signaling Analysis

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The antibodies anti-H3K9ac (#2594), anti-H3K27ac (#8173), anti-H3K56ac (#07-677-1S), anti-H4K8ac (#2594), anti-H4K12ac (#13944), anti-H4K16ac (#13534), anti-H3 (#4499), anti-H4 (#13919), anti-BRD4 (#13440), anti-Tom20 (#42406), anti-γ-H2AX (#80312), anti-H2AX (#7631), anti-p-TBK1(Cell #5483), anti-TBK1 (#3504), anti-p-IRF3 (#29047y), anti-IRF3 (#11904), anti-p-P65 (#3033), anti-P65 (#8242), anti-p-STAT1(#9167), anti-STAT1(#9172), anti-STING (#3337) and anti-p-P53 (#9286) were from Cell Signaling Technology; anti-cGAS (26416–1-AP), anti-P53 (10442-1-AP), anti-MDM2 (19058-1-AP) and anti-P21 (#10355-1-AP) were from Proteintech; anti-dsDNA (MAB1293) was from Millipore; and anti-actin (A1978) was from Sigma. Antiserum against PRV gE was generated by immunization of mice with purified recombinant gE.
Wang J., Li G.L., Ming S.L., Wang C.F., Shi L.J., Su B.Q., Wu H.T., Zeng L., Han Y.Q., Liu Z.H., Jiang D.W., Du Y.K., Li X.D., Zhang G.P., Yang G.Y, & Chu B.B. (2020). BRD4 inhibition exerts anti-viral activity through DNA damage-dependent innate immune responses. PLoS Pathogens, 16(3), e1008429.
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4

Comprehensive Antibody Characterization for Epigenetic Studies

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Antibodies used for Western blot, immunoflurorescence, and ChIP are as follows: anti-H3K27me3 (Cell Signaling #9756, 1:100 for IF, i.e. immunofluorescence), anti-H2A (Cell Signaling #2578, 1:300 for WB, i.e. Western blotting), anti-H4 (Cell Signaling #2935, 1:800 for WB), anti-γ-H2A.X (Cell signaling #9718, 1:200 for IF; 1:1500 for WB), anti-H3K9me3 (Active Motif #39385, 1:100 for IF), anti-H1 (Active Motif #39707, 1:800 for WB), anti-H2B (Active Motif #39125, 1:800 for WB), anti-H4Ac-pan (Active Motif #39243, 1:1000 for WB; 1:200 for IF), anti-H3 (Abcam #ab1791, 1:15000 for WB), anti-Chd5 (Santa Cruz Biotechnology #sc-68389, 1:1500 for WB; 1:200 for IF), anti β-Actin (Sigma #A2228, 1:2000 for WB), anti-H4K5/8/12/14Ac (Millipore #05-1335, 1:100 for IF), anti-lectin PNA Alex Fluor 568 (Invitrogen #L32458, 1:4000 for IF), anti-Prm1 (Briar Patch Biosciences, Hup1N, 1:300 for WB; 1:150 for IF), antiPrm2 (Briar Patch Biosciences, Hup2B, 1:800 for WB; 1: 200 for IF), anti-Tnp1 (gift from Dr. Stephen Kistler, University of South Carolina, 1:500 for WB; 1: 100 for IF), anti-Tnp2 (gift from Dr. Stephen Kistler, University of South Carolina, 1:1000 for WB; 1: 200 for IF) anti-nucleosome (mab #32, gift from Dr. Jo H.M. Berden, Radboud University Nijmegen Medical Center, 1:300 for IF).
Li W., Wu J., Kim S.Y., Zhao M., Hearn S.A., Zhang M.Q., Meistrich M.L, & Mills A.A. (2014). Chd5 orchestrates chromatin remodeling during sperm development. Nature communications, 5, 3812.
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5

Protein Extraction and Analysis for Cellular Studies

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For total protein, cells were lysed in radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor mixture (Roche Applied Science, Mannheim, Germany). The lysates were harvested and centrifuged for 30 min at 12000 rpm and 4 °C. The Pierce BCA protein assay kit (Thermo Scientific) was utilized to detect the protein concentrations of the supernatants. Histone protein was extracted using the EpiQuik Total Histone Extraction Kit according to the manufacturer’s protocol (Epigentek, USA). Aliquots (20 μg) of the protein extracts were separated by 10% SDS PAGE and transferred to polyvinylidene difluoride membrane (Merck Millipore, Darmstadt, Germany). The membranes were blocked with 5% skim milk for 1 h and incubated with primary antibodies at 4 °C overnight. The ECL kit (CWBIO) was used to detect immunoreactive protein bands after incubation for 1 h with peroxidase-linked secondary antibodies at room temperature. The antibodies used were: anti-GAPDH, anti-PRMT3, anti-OCN (Abcam, Cambridge, UK), anti-H4, anti-RUNX2, anti-Flag (Cell Signaling, Danvers, MA, USA), and anti-H4R3me2a (Active Motif, Carlsbad, CA).
Min Z., Xiaomeng L., Zheng L., Yangge D., Xuejiao L., Longwei L., Xiao Z., Yunsong L., Ping Z, & Yongsheng Z. (2019). Asymmetrical methyltransferase PRMT3 regulates human mesenchymal stem cell osteogenesis via miR-3648. Cell Death & Disease, 10(8), 581.
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