Rabbit anti h3k27ac

Manufactured by Abcam

Rabbit anti-H3K27ac is a primary antibody that specifically recognizes acetylated histone H3 at lysine 27 (H3K27ac). H3K27ac is a histone modification associated with active enhancers and promoters.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

H3K27ac (231 citations) Anti-H3K27ac (108 citations) Rabbit anti- (5 citations) Rabbit polyclonal anti-Histone H3K27ac (3 citations)

8 protocols using rabbit anti h3k27ac

Histone Extraction and Western Blot

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

Whole retina was dissected and homogenized in a dounce homogenizer and histones isolated with the Histone Extraction Kit (ab113476, Abcam). Extracts were run on a 4–20% gel (Bio-Rad) before blotting. PVDF membranes (Bio-Rad) were then blocked overnight at 4 °C in a milk blocking solution (5% powdered milk, 0.5% Triton X-100, 0.1% NaN₃ in PBS). Following three 30-min washes in a wash buffer (0.5% Tween in PBS), membranes were incubated with primary antibodies in milk blocking solution overnight at 4 °C. Membranes were then washed three times with wash buffer and incubated with at room temperature with 1:10,000 rabbit anti-HRP in TBS-T for 2 h. Following a final wash step, membranes were developed with SuperSignal West Dura Extended Duration Substrate (Thermo Scientific) and exposed on Autoradiography Film (Blue Devil West). All blots were scanned at 600 DPI and quantified in ImageJ (NIH, Bethesda, MD) by transforming the image to 8-bit format and quantifying the grey value via Uncalibrated OD in a region of interest drawn around histone bands.
Primary antibodies: rabbit anti-Histone H3 (Cell Signaling, 1:10,000), rabbit anti-H3K27ac (Abcam, 1:10,000).

Jorstad N.L., Wilken M.S., Grimes W.N., Wohl S.G., VandenBosch L.S., Yoshimatsu T., Wong R.O., Rieke F, & Reh T.A. (2017). Stimulation of functional neuronal regeneration from Müller glia in adult mice. Nature, 548(7665), 103-107.

+ Open protocol

+ Expand

ChIP-seq for Histone Marks and TFs

Cited 2 times

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

ChIPs were performed from 5–10×10⁶ cross-linked cells and sequencing libraries were prepared as previously described^{52 (link)}. The following antibodies were used for ChIP: rabbit anti-H3K4me1 (Abcam #8895), rabbit anti-H3K27ac (Abcam #4729), rabbit anti-c-Fos (Santa Cruz #sc-52), rabbit anti-FOSL1/Fra-1 (Santa Cruz #sc-605). ChIP-seq libraries were sequenced on the HiSeq 2000 or 2500 platform at the Case Western Reserve University Genomics Core Facility.
Analysis was performed as previously described^{8 (link)}.

Morrow J.J., Bayles I., Funnell A.P., Miller T.E., Saiakhova A., Lizardo M.M., Bartels C.F., Kapteijn M.Y., Hung S., Mendoza A., Dhillon G., Chee D.R., Myers J.T., Allen F., Gambarotti M., Righi A., DiFeo A., Rubin B.P., Huang A.Y., Meltzer P.S., Helman L.J., Picci P., Versteeg H., Stamatoyannopolus J., Khanna C, & Scacheri P.C. (2018). Positively selected enhancer elements endow osteosarcoma cells with metastatic competence. Nature medicine, 24(2), 176-185.

+ Open protocol

+ Expand

KSHV ChIP-seq Assays with Epigenetic Modifications

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

ChIP assays were performed as described previously with minor modifications [73 (link)]. Briefly, KSHV iSLK or BCBL1 cells were mock treated or treated with 3.5% 1, 6-HD for 1 hr. The cells (~ 1 x10⁷) were crosslinked in 1% formaldehyde for 15 min with vigorous shaking at room temperature, quenched in 0.125 M glycine for 5 min, and washed two times with PBS prior to ChIP lysis. For ChIP in DAXX depletion, iSLK cells were infected with lentiviruses encoding shDAXX or shControl. At 48 hrs post-infection, cells were expanded and cultured under puromycin (1 μg/ml) selection for 4 days. At day 6 post-infection, the selected cells were collected and subject to ChIP analysis. The antibodies used in ChIP assays include rat polyclonal anti-HHV8 or anti-LANA [LN53] (Abcam, ab4103), rabbit anti-H3K4me3 (EMD Millipore, 07–473), rabbit anti-H3K9me3 (Diagenode, C15410056), rabbit anti-H3K27me3 (Active Motif, 39155), rabbit anti-H3K27Ac (Abcam, ab4729), rabbit anti-H3 (EMD Millipore, 07–690), and rabbit IgG (Santa Cruz Biotechnology).

Vladimirova O., De Leo A., Deng Z., Wiedmer A., Hayden J, & Lieberman P.M. (2021). Phase separation and DAXX redistribution contribute to LANA nuclear body and KSHV genome dynamics during latency and reactivation. PLoS Pathogens, 17(1), e1009231.

+ Open protocol

+ Expand

ChIP-seq for Histone Marks and TFs

Cited 2 times

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

+ Open protocol

+ Expand

ChIP-seq Analysis of Histone Marks

Cited 1 time

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

The ChIP was carried out in accordance with a previously described protocol [39 (link)]. Briefly, cells were cross-linked using 1% formaldehyde, lysed using rotation, and broken using an ultrasonic breaker (Diagenode Bioruptor Pico, Belgium). MAGnify Chromatin Immunoprecipitation System (Thermo Fisher Scientific) was used to perform immunoprecipitation (IP) as per the manufacturer’s instructions with the antibody rabbit anti-H3K27ac (Abcam, Ab4729) or rabbit anti-H3K27me3 (Millipore, 07-449). Chromatin samples were sent for high-throughput sequencing. We used Bowtie2 (v2.3.1) to align the chromatin immunoprecipitation sequence (ChIP-seq) DNA reads to the reference genome. Each ChIP library was compared against the DNA input background library in the corresponding cell type condition. Specifically, histone marks H3K27me3 and H3K27ac in Usp7 FGSCs and Usp7-knockdown FGSC control were compared with the DNA input Usp7-knockdown FGSCs and Usp7-knockdown FGSC control, respectively. FDR < 0.05 was used to determine ChIP-seq peaks relative to the input libraries.

Zhao Y., Li X., Tian G., Zhao X., Wong J., Shen Y, & Wu J. (2020). Ubiquitin-Specific-Processing Protease 7 Regulates Female Germline Stem Cell Self-Renewal Through DNA Methylation. Stem Cell Reviews and Reports, 17(3), 938-951.

+ Open protocol

+ Expand

Profiling Histone Modifications and Transcription Factors via AutoCUT&RUN

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

We used Rabbit anti-CTCF (1:100, Millipore Cat#07-729), Rabbit anti-NPAT (1:100, Termo Fisher Cat#PA5-66839), Rabbit anti-H3K4me1 (1:100, Abcam Cat#ab8895), Rabbit anti-H3K4me2 (1:100, Millipore Cat#07-030), Rabbit anti-H3K4me3 (1:100, Active Motif Cat#39159), Rabbit anti-H3K27me3 (1:100, Cell Signaling Tech Cat#9733S). Since pA-MNase does not bind efficiently to many mouse antibodies, we used a rabbit anti-Mouse IgG (1:100, Abcam, Cat#ab46540) as an adapter. H3K27ac was profiled by AutoCUT&RUN in H1 and K562 cells and manually in VUMC-10 and SU-DIPG-XIII cell lines using Rabbit anti-H3K27ac (1:50, Millipore Cat#MABE647). H3K27ac was profiled by AutoCUT&RUN in VUMC-10 and SU-DIPG-XIII cell lines and xenografts using Rabbit anti-H3K27ac (1:100, Abcam Cat#ab45173).

Janssens D.H., Wu S.J., Sarthy J.F., Meers M.P., Myers C.H., Olson J.M., Ahmad K, & Henikoff S. (2018). Automated in situ chromatin profiling efficiently resolves cell types and gene regulatory programs. Epigenetics & Chromatin, 11, 74.

+ Open protocol

+ Expand

Antibody-based Protein Expression Analysis

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

Following antibodies were used: Mouse anti-NRG1 (Santa Cruz Biotechnology) (sc-28916; 1:1000 for blotting); Mouse anti-ErbB4 (Santa Cruz Biotechnology) (sc-8050; 1:1000 for blotting); Rabbit anti-P-ErbB4 (Cell Signaling Technology) (Tyr1284; 1:1000 for blotting); Mouse anti-GAPDH (Santa Cruz Biotechnology) (sc-32233; 1:1000 for blotting); Rabbit anti-H3 (Active motive) (61799; 1:1000 for blotting); Rabbit anti-H3k18ac (Active motive) (39587; 1:1000 for blotting); Rabbit anti-H3k27ac (Active motive) (39133; 1:1000 for blotting); Rabbit anti-H4 (Active motive) (61299; 1:1000 for blotting); Rabbit anti-H4k8ac (Active motive) (61103; 1:1000 for blotting); Rabbit anti-H4k12ac (Active motive) (39165; 1:1000 for blotting);); Rabbit anti-acetylated H4 (abcam) (ab233193; 1:200 for CHIP). Rabbit anti-acetylated H3 (Sigma) (06599; 1:200 for CHIP).
Unless otherwise indicated, chemicals were purchased from Sigma-Aldrich. DL-AP5 (0105), CNQX (0190) were purchased from Tocris Bioscience. 1NM-PP1 (HY-13942) and ITSA-1 (HY-100508) were from MedChemExpress.

Wang J., Huang J., Yao S., Wu J.H., Li H.B., Gao F., Wang Y., Huang G.B., You Q.L., Li J., Chen X, & Sun X.D. (2021). The ketogenic diet increases Neuregulin 1 expression via elevating histone acetylation and its anti-seizure effect requires ErbB4 kinase activity. Cell & Bioscience, 11, 93.

+ Open protocol

+ Expand

Chromatin Immunoprecipitation Sequencing (ChIP-seq)

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

ChIPs were performed using 2 × 10⁶ to 10 × 10⁶ cross‐linked cells, and sequencing libraries were prepared as previously described. The following antibodies were used for ChIP: rabbit anti‐H3K4me3 (Abcam), rabbit anti‐H3K27ac (Abcam), rabbit anti‐H3K27me3 (CST), and rabbit anti‐HA (CST). ChIP‐seq libraries were sequenced on the HiSeq 2000 platform at the Novogene LLC. Analysis was performed as previously described.

Lu B., Zou C., Yang M., He Y., He J., Zhang C., Chen S., Yu J., Liu K.Y., Cao Q, & Zhao W. (2021). Pharmacological Inhibition of Core Regulatory Circuitry Liquid–liquid Phase Separation Suppresses Metastasis and Chemoresistance in Osteosarcoma. Advanced Science, 8(20), 2101895.

+ 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!