H3k27ac

Manufactured by Cell Signaling Technology

Sourced in United States, Japan

H3K27ac is an antibody that specifically recognizes the acetylation of lysine 27 on histone H3. It is a useful tool for studying histone modifications and their roles in various cellular processes.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Antibody against H3K27ac Acetyl-histone H3 Lys27 (H3K27Ac, H3K27ac (8173S) Anti-H3K27ac Rabbit anti-H3K27ac H3K27Ac antibody Histone H3K27ac Anti-H3K27Ac antibody Anti‐H3K27ac (D5E4) Anti-histone H3K27ac

74 protocols using h3k27ac

Antibody sources for protein detection

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The RING1B mouse monoclonal antibody has been described previously (Atsuta et al., 2001 (link)). A rabbit polyclonal antibody against the Flag-2XStrepII sequence was generated as described previously (Farcas et al., 2012 (link)). Commercially available antibodies were used to detect FLAG-tag (Sigma-Aldrich, St. Louis, MO,, F1804, RRID:AB_262044), HA-tag (Santa Cruz, Dallas, TX, sc-805), RYBP (Millipore, Billerica, MA, AB3637, RRID:AB_631618), L3MBTL2 (Active Motif, Carlsbad, CA, 39569, RRID:AB_2615062), MAX (Santa Cruz, sc-197X, RRID:AB_2281783), MGA (Bethyl, Montgomery, TX, A302-864A, RRID:AB_2615457), PCGF1 (Santa Cruz, E-8: sc-515371), PCGF2 (Santa Cruz, H-115: sc-10744, RRID:AB_2267885), PCGF6 (ORIGENE, Rockville, MD, TA324658), EZH2 (Cell Signaling, Danvers, MA, 4905, RRID:AB_2278249), SUZ12 (Cell Signaling, 3737, RRID:AB_2196850), SET1 (Bethyl, A300-289A, RRID:AB_263413), trimethylated Histone H3 lysine 27 (H3K27me3) (Millipore, 07–449, RRID:AB_310624), dimethylated histone H3 lysine 9 (H3K9me2) (MBL, Japan, MABI0317), H3K27ac (Cell Signaling, 8173, RRID:AB_2616015), monoubiquitinated histone H2A lysine 119 (H2AK119ub1) (Millipore, 05–678, RRID:AB_309899; Cell Signaling, 8240, RRID:AB_10891618), mouse IgM (Millipore, 12–488, RRID:AB_390193), histone H3 (Millipore, 07–690, RRID:AB_417398), and LAMIN B (Santa Cruz, sc-6216, RRID:AB_648156).

Endoh M., Endo T.A., Shinga J., Hayashi K., Farcas A., Ma K.W., Ito S., Sharif J., Endoh T., Onaga N., Nakayama M., Ishikura T., Masui O., Kessler B.M., Suda T., Ohara O., Okuda A., Klose R, & Koseki H. (2017). PCGF6-PRC1 suppresses premature differentiation of mouse embryonic stem cells by regulating germ cell-related genes. eLife, 6, e21064.

+ Open protocol

+ Expand

Western Blot and ChIP Antibody Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

The primary antibodies for western blot were: HA (901501, BioLegend, dilution 1:5000), BRD4 (ab128874, Abcam, dilution 1:2500), α-Tubulin (ab7291, Abcam, dilution 1:10000), Lamin B1 (ab16048, Abcam, dilution 1:2500), Histone H3 (ab18521, Abcam, dilution 1:2500), H3K27ac (8173, Cell Signaling, dilution 1:2500), Pan-acetyl H3 (39140, Active Motif, dilution 1:2500), MYC (sc-40, Santa Cruz, dilution 1:500) and GAPDH (sc-365062, Santa Cruz, dilution 1:10000). The secondary antibodies used were goat anti-mouse HRP (115-035-003, Jackson ImmunoResearch, dilution 1:5000), goat anti-rabbit HRP (111-035-144, Jackson ImmunoResearch, dilution 1:5000). Antibodies used for ChIP-seq and ChIP-qPCR were: BRD4 (ab128874, Abcam) and rabbit polyclonal IgG (sc-66931, Santa Cruz).

Li K.C., Girardi E., Kartnig F., Grosche S., Pemovska T., Bigenzahn J.W., Goldmann U., Sedlyarov V., Bensimon A., Schick S., Lin J.M., Gürtl B., Reil D., Klavins K., Kubicek S., Sdelci S, & Superti-Furga G. (2021). Cell-surface SLC nucleoside transporters and purine levels modulate BRD4-dependent chromatin states. Nature metabolism, 3(5), 651-664.

+ Open protocol

+ Expand

ChIPmentation for MYCN and H3K27ac

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies for ChIP using tagmentation (ChIPmentation) were as follows: MYCN (catalog sc-791; Santa Cruz Biotechnology Inc.); H3K27ac (catalog 8173S; Cell Signaling Technology). ChIPmentation was performed as previously described (53 (link)). ChIPmentation libraries were sequenced on a NextSeq 500 (Illumina).

Poon E., Liang T., Jamin Y., Walz S., Kwok C., Hakkert A., Barker K., Urban Z., Thway K., Zeid R., Hallsworth A., Box G., Ebus M.E., Licciardello M.P., Sbirkov Y., Lazaro G., Calton E., Costa B.M., Valenti M., De Haven Brandon A., Webber H., Tardif N., Almeida G.S., Christova R., Boysen G., Richards M.W., Barone G., Ford A., Bayliss R., Clarke P.A., De Bono J., Gray N.S., Blagg J., Robinson S.P., Eccles S.A., Zheleva D., Bradner J.E., Molenaar J., Vivanco I., Eilers M., Workman P., Lin C.Y, & Chesler L. (2020). Orally bioavailable CDK9/2 inhibitor shows mechanism-based therapeutic potential in MYCN-driven neuroblastoma. The Journal of Clinical Investigation, 130(11), 5875-5892.

+ Open protocol

+ Expand

Histopathological Evaluation of Tumour Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

Formalin-fixed, paraffin-embedded tumour sections were analysed by histology after generating 4-micrometre tissue sections and staining with haematoxylin and eosin. Immunohistochemical stains were performed on representative tissue sections using H3K27Ac (Cell Signaling Technology, no. 8173), H3K27Me3 (Cell Signaling Technology, no. 9733), EZH2 (Cell Signaling Technology, no. 5246), BAF47 (SMARCB1) (BD Transduction Laboratories, no. 612110), Cleaved caspase-3 (Cell Signaling Technology, no. 9664S) and PCNA (Cell Signaling Technology, no. 2586S). Following sodium citrate (pH 6.0) antigen retrieval, staining was performed using the Vectastain ABC Elite Rabbit IgG Kit, ABC Elite Mouse IgG Kit or M.O.M^TM Kit (Vector Laboratories, Burlingame, United States of America) according to manufacturer’s instructions. Slides were counterstained with haematoxylin and imaged using the Nikon ECLIPSE TS100 bright-field microscope (Nikon, Minato City, Japan).

Chong W.C., Jayasekara W.S., Vaghjiani V.G., Parackal S., Sun C., Popovski D., Algar E.M., Firestein R., Wood P.J., Khan S., Huang A., Ashley D.M., Downie P, & Cain J.E. (2021). Atypical Teratoid Rhabdoid Tumours Are Susceptible to Panobinostat-Mediated Differentiation Therapy. Cancers, 13(20), 5145.

+ Open protocol

+ Expand

Western Blot Analysis of Mitochondrial and Epigenetic Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blot was performed as described^{2 (link)} using the following antibodies: TFAM (7495S, Cell Signaling Technology; sc-23588, Santa Cruz Technology), PHB2 (14085S, Cell Signaling Technology), ATP5A1 (sc-136178, Santa Cruz Technology), ATP5B (sc-16690, Santa Cruz Technology), ATP5D (ab97491, Abcam), ATP5O (ab110276, Abcam, clone 4C11C10D12), 4EBP1 (9644S, Cell Signaling Technology), phos-4EBP1 (Thr37/46) (2855S, Cell Signaling Technology), p70S6K (2708S, Cell Signaling Technology), phos-S6K (Ser235/236) (2211S, Cell Signaling Technology), GATA1 (ab47490, Abcam), GATA2 (ab22849, Abcam), TSC1 (6935, Cell Signaling Technology), TSC2 (4308, Cell Signaling Technology), H3K9ac (9649, Cell Signaling Technology), H3K27ac (8173, Cell Signaling Technology), H3K56ac (4243, Cell Signaling Technology), H4K5ac (ab51997, Abcam), H3K4me3 (9571, Cell Signaling Technology), H3K9me3 (13969, Cell Signaling Technology), H3K27me3 (9733, Cell Signaling Technology), H3K36me3 (4909, Cell Signaling Technology), H3 (4499, Cell Signaling Technology), ACTB (MAB1501, Millipore, clone C4), and GAPDH (sc-26778, Santa Cruz Biotechnology). Densitometry quantification was performed using ImageJ software. All antibodies were used at 1:1,000 dilutions except phos-4EBP1 (1:500), phos-S6K (1:500) and ACTB (1:2,500).

Liu X., Zhang Y., Ni M., Cao H., Signer R.A., Li D., Li M., Gu Z., Hu Z., Dickerson K.E., Weinberg S.E., Chandel N.S., DeBerardinis R.J., Zhou F., Shao Z, & Xu J. (2017). Regulation of Mitochondrial Biogenesis in Erythropoiesis by mTORC1-Mediated Protein Translation. Nature cell biology, 19(6), 626-638.

+ Open protocol

+ Expand

Measuring Histone Modifications and Phosphorylation by Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were harvested and washed twice with PBS. For H3K27ac measurement, cells were then treated with 4% paraformaldehyde in PBS for 15 minutes on ice, followed by centrifugation for 5 minutes at 300 g. Next, cells were resuspended in cold 70% EtOH added drop by drop while vortexing gently and then incubated overnight at −20°C. Then, cells were washed twice with PBS and once with PBS/1% FBS. Cells were permeabilized for 20 minutes with 0.25% Triton X-100 in PBS/1% FBS followed by incubation with primary H3K27ac (Cell Signaling Technology 8173S) or phospho-Akt (Cell Signaling Technologies, 4060S) antibodies at 1:500 dilution for 30 minutes, and then secondary antibody (1:1000) (Life Technologies Goat anti-rabbit IgG cross-adsorbed antibody Alexa Fluor 555, A21428) for 2 hours at 4°C in the dark, prior to flow cytometry analysis.

Mowery C.T., Reyes J.M., Cabal-Hierro L., Higby K.J., Karlin K.L., Wang J.H., Kimmerling R.J., Cejas P., Lim K., Li H., Furusawa T., Long H.W., Pellman D., Chapuy B., Bustin M., Manalis S.R., Westbrook T.F., Lin C.Y, & Lane A.A. (2018). Trisomy of a Down Syndrome Critical Region Globally Amplifies Transcription via HMGN1 Overexpression. Cell reports, 25(7), 1898-1911.e5.

+ Open protocol

+ Expand

Evaluation of CDK12 and RNAPII Inhibitors

Check if the same lab product or an alternative is used in the 5 most similar protocols

THZ531 (HY-103618), enzalutamide (HY-70002), apalutamide (HY-16060), and bicalutamide (HY-14249) were purchased from MCE. Antibodies against CDK12 (ab37914) were purchased from Abcam. Additional antibodies against CDK12 (PAB39156) were purchased from Bioswamp. Antibodies against phospho-CTD-RNAPII-S2 (Cat# 041571-l, lot# 3023013), CTD-RNAPII (Cat# 05–623–25UG, lot# Q2925497), AR (Cat# 5153T), and H3K27ac (Cat# 8173T, lot# 6) were purchased from Cell Signaling Technology. Antibodies against GRIN3A (Cat# bs12100R) were purchased from Bioss.

Lei H., Wang Z., Jiang D., Liu F., Liu M., Lei X., Yang Y., He B., Yan M., Huang H., Liu Q, & Pang J. (2021). CRISPR screening identifies CDK12 as a conservative vulnerability of prostate cancer. Cell Death & Disease, 12(8), 740.

+ Open protocol

+ Expand

Comprehensive Immunohistochemistry Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

The antibodies used were: ASCL1 (Abcam, #ab211327), INSM1 (Abcam, #ab170876), E2F7 (Abcam, ab245655), RB1 (Abcam, ab181616), SYP (Santa Cruz, #sc-17750), RCOR1 (Santa Cruz, #sc-376567), CCN1 (Santa Cruz, #sc-374129), CCN2 (Santa Cruz, #sc-365970), YAP/TAZ (Santa Cruz, #sc-101199), P53 (Santa Cruz, #sc-126), E2F1 (Santa Cruz, #sc-251), GAPDH (Santa Cruz, #sc-47724), LATS1 (Cell signaling, #3477), LATS2 (Cell signaling, #5888), p-LATS (Cell signaling, #8654), H3 (Cell signaling, #4499), H3K27ac (Cell signaling, #8173), H3K4me3 (Cell signaling, #9751), VIN (Cell signaling, #13901), Flag (Sigma, #1804),

Wu Z., Su J., Li F.L., Chen T., Mayner J., Engler A., Ma S., Li Q, & Guan K.L. (2023). YAP silencing by RB1 mutation is essential for small-cell lung cancer metastasis. Nature Communications, 14, 5916.

+ Open protocol

+ Expand

Metaphase Arrest and Immunofluorescence

Check if the same lab product or an alternative is used in the 5 most similar protocols

We added 50 ng/ml Karyomax to the media of day 10 differentiating embryoid bodies for 4 h to arrest cells in metaphase. We harvested the cells via trypsinization, and trypsin was quenched by addition of media. We spun the cells at 1000 rpm for 5 min, aspirated the media, then washed twice in 1× PBS. Cells were then resuspended to a concentration of 5 × 10⁵ cell/ml in 75 mM KCl, and placed at 37 °C for 10 min for swelling. 1 × 10⁵ cells were then cytospun onto a microscope slide at 1000 rpm for 5 min. The cells were fixed in PFA and immunofluorescence was performed as described for interphase cells. We stained H3K27me3 with a 1:200 dilution of Active Motif 39535 and H3K27ac with a 1:200 dilution of Cell Signaling D5E4.

Froberg J.E., Pinter S.F., Kriz A.J., Jégu T, & Lee J.T. (2018). Megadomains and superloops form dynamically but are dispensable for X-chromosome inactivation and gene escape. Nature Communications, 9, 5004.

+ Open protocol

+ Expand

ChIP-seq Analysis of Rat Schwann Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

ChIP assays were performed with minor modifications^{43 (link)}. Purified rat SCs were fixed for 15 min at room temperature with 1% formaldehyde-containing medium. Nuclei were isolated and sonicated in sonication buffer (10 mM Tris-HCl pH 8.0, 1 mM EDTA, 0.5 mM EGTA and protease inhibitor cocktail). Sonicated chromatin (~300 µg) was used for immunoprecipitation by incubation with appropriate antibodies (4 μg) overnight at 4 °C. Prerinsed magnetic protein A/G beads (50 μL, Thermo Fisher Scientific, 26162) were added to each ChIP reaction and reactions were incubated for 1 h at 4 °C. The beads were then incubated in 200 ml elution buffer at 65 °C for 20 min to elute immunoprecipitated materials. The ChIP-seq libraries were prepared using NEBNext ChIP-seq Library Prep Master Mix Set for Illumina (NEB catalogue number E6240L) and then run on the Illumina sequencer HiSeq 2500. We used antibodies CTCF (rabbit, Cell Signaling, #3418), H3K27Ac (rabbit, Active motif, 39135), or H3K27me3 (rabbit, Cell Signaling, 9733 s) for ChIP. The crosslinked and sonicated chromatins without immunoprecipitation were used as input controls. For ChIP, real-time PCR was performed using quantitative SYBR green PCR mix (Bio-Rad, 1725121). The values of IgG were normalized to 1.

Wang J., Wang J., Yang L., Zhao C., Wu L.N., Xu L., Zhang F., Weng Q., Wegner M, & Lu Q.R. (2020). CTCF-mediated chromatin looping in EGR2 regulation and SUZ12 recruitment critical for peripheral myelination and repair. Nature Communications, 11, 4133.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!