Anti h2ak119ub d27c4

Manufactured by Cell Signaling Technology

Anti-H2AK119ub (D27C4) is a rabbit monoclonal antibody that recognizes the histone H2A monoubiquitination at lysine 119 (H2AK119ub). This antibody is designed for the detection of H2AK119ub in various applications such as Western blotting, immunoprecipitation, and immunofluorescence.

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Lab products found in correlation

Mowiol 4 88, by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole dihydrochloride, by Merck Group (1 mentions) Sp8 confocal microscope, by Leica (1 mentions) Cy 3 affinipure donkey anti rabbit igg h l, by Jackson ImmunoResearch (1 mentions) Anti h3k4me3, by Diagenode (1 mentions) Anti h3k27ac, by Diagenode (1 mentions) Anti ezh2 d2c9, by Cell Signaling Technology (1 mentions) Anti h3k4me1, by Diagenode (1 mentions) Anti cbx8, by Diagenode (1 mentions) Anti h3k27me3, by Diagenode (1 mentions) Goat anti rabbit immunoglobulins hrp, by Agilent Technologies (1 mentions) Odyssey clx imaging system, by LI COR (1 mentions) Anti mdm2 sc 813, by Santa Cruz Biotechnology (1 mentions) Rabbit anti mouse immunoglobulins hrp, by Agilent Technologies (1 mentions) Ab183499, by Abcam (1 mentions) Anti β actin c4, by Santa Cruz Biotechnology (1 mentions) Chemidoc xrs system, by Bio-Rad (1 mentions) Ab181140, by Abcam (1 mentions) Anti gfp sc 9996, by Santa Cruz Biotechnology (1 mentions) Anti gfp ab290, by Abcam (1 mentions)

Spelling variants of this product

H2AK119ub (22 citations) Anti-H2AK119ub (10 citations) H2AK119ub (D27C4, (3 citations)

3 protocols using anti h2ak119ub d27c4

Skin Tissue Preparation for Immunofluorescence

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Back skin was harvested at the described time points, fixed with 4% paraformaldehyde (PFA) for 3 hours at 4°C, and cryopreserved in 30% sucrose overnight, followed by O.C.T. embedding (Tissue-TEK 4583). Embedded tissues were cut at 7 μm in thickness. Sections were washed in tris-buffered saline–0.1% Tween 20 (TBS-T) and blocked with 5% donkey serum at room temperature for 1 hour. Section were incubated with anti-H2AK119Ub (D27C4, Cell Signaling Technology) overnight at 4°C, washed in TBS-T, incubated for 1 hour at room temperature with secondary antibody [715-165-147, Cy3 AffiniPure Donkey Anti-Rabbit IgG (H+L), Jackson ImmunoResearch] and 4′,6-diamidino-2-phenylindole dihydrochloride (32670, Sigma-Aldrich), and mounted with Mowiol 4-88 (81381, Sigma-Aldrich). Images were taken with Leica Sp8 confocal microscope.

Pivetti S., Fernandez-Perez D., D’Ambrosio A., Barbieri C.M., Manganaro D., Rossi A., Barnabei L., Zanotti M., Scelfo A., Chiacchiera F, & Pasini D. (2019). Loss of PRC1 activity in different stem cell compartments activates a common transcriptional program with cell type–dependent outcomes. Science Advances, 5(5), eaav1594.

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ChIP Analysis Protocol for Epigenetic Markers

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ChIP analysis was essentially performed as described previously (Frank et al., 2001 (link)). ChIP reactions were performed using the following antibodies: anti-GFP (ab290, Abcam), anti-CBX8 (C15410333, Diagenode), anti-EZH2 (D2C9, Cell Signaling), anti-H2AK119ub (D27C4, Cell Signaling), anti-H3K27me3 (C15410195, Diagenode), anti-H3K27Ac (C15410196, Diagenode), anti-H3K4me1 (C15410194, Diagenode), and anti-H3K4me3 (C15410003, Diagenode). ChIP efficiencies were assessed using qPCR. Primer sequences can be found in Supplementary file 6.

Azkanaz M., Rodríguez López A., de Boer B., Huiting W., Angrand P.O., Vellenga E., Kampinga H.H., Bergink S., Martens J.H., Schuringa J.J, & van den Boom V. (2019). Protein quality control in the nucleolus safeguards recovery of epigenetic regulators after heat shock. eLife, 8, e45205.

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Comprehensive Western Blot Analysis

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Western blotting was performed as previously described (van den Boom et al., 2013 (link)). The following primary antibodies were used in this study: anti-KDM2B (09-864, Merck), anti-USP7 (A300-033A, Bethyl Laboratories), anti-TRIM27 (18791, IBL), anti-RING1A (ab180170, Abcam), anti-RING1B (ab181140, Abcam), anti-PCGF1 (ab183499, Abcam), anti-STAT5 (sc-835-G, Santa Cruz Biotechnology), anti-TP53 (sc-216, Santa Cruz Biotechnology), anti-MDM2 (sc-813, Santa Cruz Biotechnology), anti-GFP (sc-9996, Santa Cruz Biotechnology), anti-GFP (ab290, Abcam), anti-Ubiquitin (FK2, Enzo Life Sciences), anti-H2AK119ub (D27C4, Cell Signaling), and anti-b-Actin (C4, Santa Cruz Biotechnology). Secondary antibodies used included either goat anti-mouse IRDye 800 or goat anti-rabbit IgG (H+L) Alexa Fluor 680 (Invitrogen) for imaging using an Odyssey CLx Imaging System (Li-Cor Biosciences), or goat anti-rabbit immunoglobulins/HRP (Agilent-Dako), and rabbit anti-mouse immunoglobulins/HRP (Agilent-Dako) for imaging using a ChemiDoc XRS+ System (Biorad).

Maat H., Atsma T.J., Hogeling S.M., Rodríguez López A., Jaques J., Olthuis M., de Vries M.P., Gravesteijn C., Brouwers-Vos A.Z., van der Meer N., Datema S., Salzbrunn J., Huls G., Baas R., Martens J.H., van den Boom V, & Schuringa J.J. (2021). The USP7-TRIM27 axis mediates non-canonical PRC1.1 function and is a druggable target in leukemia. iScience, 24(5), 102435.

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