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Anti h3

Manufactured by Abcam
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Anti-H3 is a primary antibody that recognizes the histone H3 protein. Histone H3 is a core component of nucleosomes, the fundamental units of chromatin. This antibody can be used for various applications, such as Western blotting, immunoprecipitation, and chromatin immunoprecipitation (ChIP).

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Lab products found in correlation

Anti h3k4me3, by Abcam (13 mentions) Anti flag, by Merck Group (11 mentions) Anti h3k9me3, by Abcam (8 mentions) Anti gapdh, by Merck Group (8 mentions) Anti h3k27me3, by Cell Signaling Technology (8 mentions) Anti h3k36me3, by Abcam (7 mentions) Anti h3k4me3, by Merck Group (7 mentions) Anti tubulin, by Abcam (7 mentions) Anti α tubulin, by Merck Group (7 mentions) Ab1791, by Abcam (7 mentions) Anti h3k9me2, by Abcam (6 mentions) Anti lsd1, by Abcam (6 mentions) Anti h3k27me3, by Merck Group (6 mentions) Anti h3k27me3, by Abcam (6 mentions) Anti β actin, by Merck Group (6 mentions) Anti h3k4me3, by Active Motif (5 mentions) Anti h3k9ac, by Merck Group (5 mentions) Anti h3k4me2, by Merck Group (5 mentions) Anti ezh2, by Cell Signaling Technology (4 mentions) Anti h3k27me3, by Diagenode (4 mentions)

Spelling variants of this product

Mouse anti-histone H3 (11 citations) Anti-phospho-histone H3 (8 citations) Anti-Histone H3 (tri methyl K27) antibody (5 citations) Anti-Histone H3 (di methyl K4) antibody (3 citations) Anti-histone H3 (acetyl K9) antibody (3 citations) Anti-human B7-H3 (2 citations) Rabbit anti-histone H3 (acetyl K27) (2 citations) Anti-citrullinated histone H3 (ab5103, (2 citations) Rabbit polyclonal anti‐Histone H3 antibody (ab1791), (2 citations) Anti-acetyl-Histone H3 antibody (2 citations)

148 protocols using anti h3

1

Quantitative Analysis of Chromatin Modifications

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Cells were lysed in ice-cold lysis buffer (20 mM Tris–HCl pH 8, 150 mM NaCl, 10 mM MgCl2, 1 mM EDTA, 0.5% Triton X-100 + protease inhibitors) for 15 min at 4 °C. Samples were then sonicated in a Q800 Waterbath sonicator (QSonica) for 15 cycles of 8 s ON, 15 s OFF. Lysates were cleared after 15 min centrifugation at 10,000 g, and protein concentration was determined using Pierce BCA Protein assay kit (Thermo Fisher). Protein extracts were resolved by SDS-PAGE, blotted to PVDF membranes and probed using the following antibodies: anti-EZH2 (Cell Signaling Technologies, Cat nb#5246, 1:1000), anti-H3K27me3 (Diagenode, Cat nb# C15200181-50, 1:5000), anti-H3K27me2 (Abcam, Cat nb# ab24684, 1:1000), anti-H3K27me1 (Active Motif, Cat nb# 61015, 1:1000), anti-H3 (Abcam, Cat nb# ab1791, 1:2000), GAPDH (Millipore, Cat nb# MAB274, 1:10,000), actin (Abcam, Cat nb# ab3280, 1:5000). Signal was detected using IRDye 680RD and 800CW secondary antibodies following the manufacturer’s instructions (LI-COR). Membranes were acquired on an Odyssey Imager (LI-COR) and quantified using Image Studio (LI-COR). Uncropped images of all western blots are provided in the Source Data file.
Lima-Fernandes E., Murison A., da Silva Medina T., Wang Y., Ma A., Leung C., Luciani G.M., Haynes J., Pollett A., Zeller C., Duan S., Kreso A., Barsyte-Lovejoy D., Wouters B.G., Jin J., Carvalho D.D., Lupien M., Arrowsmith C.H, & O’Brien C.A. (2019). Targeting bivalency de-represses Indian Hedgehog and inhibits self-renewal of colorectal cancer-initiating cells. Nature Communications, 10, 1436.
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2

Chromatin Immunoprecipitation Protocol

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One hundred OD660 U of cells were crosslinked and sonicated in biological duplicate using the protocol described in Rodriguez et al., 2014 (link). Proteins were immunoprecipated from 1 μg chromatin and 1 μL of anti-H3 (Abcam, 1791) conjugated to 20 μL protein G magnetic beads (Invitrogen, 10004D) per reaction. For Pol II ChIPs, we used an antibody against the Rpb3 subunit (2 μL per reaction, Biolegend 665004) conjugated to 20 μL protein G magnetic beads (Invitrogen, 10004D). For Sth1 ChIP experiments, we used an antibody against the Flag-epitope tag, FLAG M2 mouse monoclonal (Sigma Aldrich, F1804) and conjugated to 20 μL protein G beads (Invitrogen, 10004D). Libraries were generated using the Ovation Ultralow v2 kit (NuGEN/Tecan, 0344) and subjected to 50 bp single-end sequencing on an Illumina HiSeq 2500 at the Fred Hutchinson Cancer Research Center genomics facility.
Cucinotta C.E., Dell R.H., Braceros K.C, & Tsukiyama T. (2021). RSC primes the quiescent genome for hypertranscription upon cell-cycle re-entry. eLife, 10, e67033.
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3

ChIP-seq Histone Modification Analysis Protocol

Cited 1 time
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ChIPs were done as previously described with the modifications described below41 (link). The following histone antibodies were used: 1 μl of anti-H3K4me2 (Upstate 07-030); 0.5 μl of anti-H3K36me3 (Abcam 9050); 0.5 μl of anti-acetyl H4 (Upstate 06-598); and 2 μl of anti-H3 (Abcam 1791). All antibodies were bound to Protein A-agarose beads and used for precipitation of formaldehyde-crosslinked chromatin. For anti-H3 antibody, binding was done in FA lysis buffer containing 275 mM NaCl. For other antibodies, binding was done in FA lysis buffer containing 1 M NaCl. Precipitated DNAs were analysed in real-time using SYBR Green Supermix and CFX96 cycler (Bio-Rad). Oligonucleotides for PCR analysis are listed in Supplementary Table 2.
Kim J.H., Lee B.B., Oh Y.M., Zhu C., Steinmetz L.M., Lee Y., Kim W.K., Lee S.B., Buratowski S, & Kim T. (2016). Modulation of mRNA and lncRNA expression dynamics by the Set2–Rpd3S pathway. Nature Communications, 7, 13534.
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4

Cardiogenic ChIP-qPCR Analysis

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For ChIP-qPCR analysis pools of 20 cardiogenic regions of E8.25-E9 embryos or 106 CPCs obtained by directed differentiation of ES cells into cardiomyocytes were used. The following antibodies were utilized: anti-Isl1 (39.4D5, DSHB), anti-Brg1 (Millipore, 07-478), anti-Gata4 (Santa Cruz, sc-1237) and anti-H3 (Abcam, ab1791) antibodies. Average threshold cycle (Ct) values were used to calculate % enrichment compared to 1% input and negative control region Ct value was used to calculate fold enrichment.
Gao R., Liang X., Cheedipudi S., Cordero J., Jiang X., Zhang Q., Caputo L., Günther S., Kuenne C., Ren Y., Bhattacharya S., Yuan X., Barreto G., Chen Y., Braun T., Evans S.M., Sun Y, & Dobreva G. (2019). Pioneering function of Isl1 in the epigenetic control of cardiomyocyte cell fate. Cell Research, 29(6), 486-501.
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5

Chromatin Immunoprecipitation Antibodies

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The antibodies used were anti-H3K4me3(Millipore, Billerica, MA 17–614), anti-H3K9me3(abcam, Cambridge, United Kingdom ab8898), anti-GFP(abcam, Cambridge, United Kingdom ab290), anti-H3(abcam, Cambridge, United Kingdom ab1791), anti-H3K9ac(Wako, Richmond, VA 309–32379).
Wang W., Chaturbedi A., Wang M., An S., Santhi Velayudhan S, & Lee S.S. (2018). SET-9 and SET-26 are H3K4me3 readers and play critical roles in germline development and longevity. eLife, 7, e34970.
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6

Epigenetic Analysis of GFP-RNase H1 in Mammalian Cells

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Transfections of GFP-RNase H1 plasmid into human HeLa and mouse embryonic fibroblasts (MEF) cells were carried out as described previously4 . Ago2 KO and parental wild type cells are MEFs. G9a/GLP double KO and their parental wild type are mouse embryonic stem (mES) cells. Treatment with 10 μM of BIX-01294 inhibitor (Sigma) was performed as described20 (link). Total RNA was isolated using TRIzol reagent (Invitrogen) and reverse transcribed with SuperScript III Reverse Transcriptase (Invitrogen) using gene-specific primers. J2 dsRNA pull-down was performed as described8 (link). RT-qPCR levels are presented graphically as raw values x1000. Chromatin immuno-precipitation (ChIP) and genomic DNA immuno-precipitation (DIP) analyses were carried out as before4 . The following antibodies were used for ChIP: anti-H3K9me2 (Abcam), anti-H3K9me3 (Abcam), anti-H3 (Abcam), anti-Dicer (13D6) (Abcam), anti-KMT1C/G9a (Abcam), anti-Ago1 (Millipore), anti-Ago2 (Abcam) and anti-Pol II (H-224) (Santa Cruz Biotechnology). S9.6 RNA:DNA hybrid specific antibody was used for DIP4
Skourti-Stathaki K., Kamieniarz-Gdula K, & Proudfoot N.J. (2014). R-loops induce repressive chromatin marks over mammalian gene terminators. Nature, 516(7531), 436-439.
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7

Western Blot Analysis of NPR1 and MAPK

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For detection of NPR1 proteins, samples of total protein (30 μg) were resolved on 8% SDS-PAGE and transferred to a PVDF membrane (Bio-Rad, United States). Primary and secondary antibodies were used rabbit anti-NPR1 (1:5,000; Abiocode, United States) antibodies and horseradish peroxidase-conjugated anti-rabbit antibodies (1:10,000), respectively. Signals were visualized using an ECL kit (Bio-Rad, United States).
For detection of MPK activities, total protein (30 μg) were resolved on 10% SDS-PAGE and transferred to a PVDF membrane (Bio-Rad, United States). Primary and secondary antibodies were used rabbit anti-phospho-p42/44 MAPK (1:2,000, Cell Signaling Technology, United States) antibodies and horseradish peroxidase-conjugated anti-rabbit antibodies (1:10,000), respectively. Signals were visualized using an ECL kit.
For detection of nuclear and cytosol proteins, nuclear and cytosolic fractions were resolved by 10% SDS-PAGE. The proteins were detected with rabbit anti-NPR1 (1:5,000; Abiocode, United States), anti-H3 (1:5,000, Abcam, United Kingdom) and anti-PEPC (1:10,000, Abcam, United Kingdom) antibodies as primary antibodies and horseradish peroxidase-conjugated anti-rabbit (1:10,000) antibodies as secondary antibodies. Signals were visualized using an ECL kit.
An J., Kim S.H., Bahk S., Vuong U.T., Nguyen N.T., Do H.L., Kim S.H, & Chung W.S. (2021). Naringenin Induces Pathogen Resistance Against Pseudomonas syringae Through the Activation of NPR1 in Arabidopsis. Frontiers in Plant Science, 12, 672552.
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8

Immunoblotting Analysis of Cellular Proteins

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Protein from mid-log phase culture was isolated by TCA (trichloroacetic acid) precipitation and resolved by electrophoresis on a SDS-polyacrylamide gel. The following primary antibodies were used for Immunoblotting: anti-HA (16B12; Covance), anti-Rad53 (EL7; gift from Pellicioli lab / Abcam, #104232), anti-Myc (Santa Cruz Biotechnology, sc-789), anti-GAPDH (Sigma, A-9521), anti tubulin (AbD serotec, MCA78G), anti-phosphorylated Rad53 (F9; gift from Pellicioli lab), anti-H3 (Abcam #1791), and anti-Ptc2 (Rabbit serum, gift from Dr. Marie-Claude Marsolier-Kergoat).
Diao L.T., Chen C.C., Dennehey B., Pal S., Wang P., Shen Z.J., Deem A, & Tyler J.K. (2017). Delineation of the role of chromatin assembly and the Rtt101Mms1 E3 ubiquitin ligase in DNA damage checkpoint recovery in budding yeast. PLoS ONE, 12(7), e0180556.
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9

Protein Extraction and Western Blot Analysis

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The total protein was extracted from 30 digestive tracts or 30 heads of 3–7 days old female adult flies using 60 μl RIPA buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 1% TritonX-100, 1% sodium deoxycholate, 0.1% SDS, 2 mM sodium pyrophosphate, 25 mM β-glycerophosphate, 1 mM EDTA, 1× Protease Inhibitor Cocktail from Roch company). The same amount of protein samples were run on the SurePAGE 4–12% or 4–20% MOPS gel (GenScript) and then transferred to polyvinylidene difluoride (PVDF) membrane. After blocking non-specific sites with 5% milk or Odyssey Blocking buffer, the membrane was incubated with the different primary antibodies including KDM5 antibody((Secombe et al., 2007 (link)), Anti-H3K4me3 (Active Motif) and Anti-H3 (Abcam) at 4 C o/n, followed by incubation with HRP-conjugated secondary antibodies (Santa cruz blotechnology) or IRDye 800CW/680RD goat anti-rabbit/goat anti-mouse secondary antibodies ((LI-COR Biosciences)). Immunoreactive bands were detected using Tanon™ High-sig ECL Western Blotting Substrate(Tanon) and visualized using ChemiDoc™ Touch Imaging System or ODYSSEY INFRARED IMAGER (LI-COR) respectively. Band intensity quantities using ImageJ.
Chen K., Luan X., Liu Q., Wang J., Chang X., Snijders A.M., Mao J.H., Secombe J., Dan Z., Chen J.H., Wang Z., Dong X., Qiu C., Chang X., Zhang D., Celniker S.E, & Liu X. (2019). Drosophila Histone Demethylase KDM5 regulates social behavior through immune control and gut microbiota maintanence. Cell host & microbe, 25(4), 537-552.e8.
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10

Immunoprecipitation and Western Blotting

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For immunoprecipitation (IP), cells were lysed in Triton lysis buffer supplemented with protein inhibitor cocktails (Thermo Fisher Scientific), followed by a brief sonication, and then lysates were immunoprecipitated with anti-E2F1 (Abcam), anti-LSD1 (Abcam), or anti-CoREST (Abcam) antibodies. For immunoblotting, proteins were separated on 4–15% SDS gradient gels (Bio-Rad), transferred to nitrocellulose membranes (Bio-Rad), and then probed with primary antibodies, including anti-Rb (Cell Signaling), anti-tubulin (Abcam), anti-LSD1 (Abcam), anti-CoREST (Abcam), anti-E2F1 (Cell Signaling), anti-GAPDH (Abcam), anti-V5 (Abcam), anti-H3 (Abcam) and anti-FOXA1 (Abcam).
Han W., Liu M., Han D., Li M., Toure A.A., Wang Z., Besschetnova A., Patalano S., Macoska J.A., Gao S., Hansen He H, & Cai C. (2022). RB1 loss in castration-resistant prostate cancer confers vulnerability to LSD1 inhibition. Oncogene, 41(6), 852-864.
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