Abe364

Manufactured by Merck Group

Sourced in United Kingdom

The ABE364 is a laboratory equipment product manufactured by Merck Group. It is designed to perform specific functions in a research or laboratory setting. The core function of the ABE364 is to [CORE FUNCTION]. Further details on the intended use or interpretation of the product's capabilities are not provided.

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Lab products found in correlation

Ab171870, by Abcam (1 mentions) Ab37355, by Abcam (1 mentions) Ab176916, by Abcam (1 mentions) H3k4me2, by Merck Group (1 mentions) H3k27me3, by Merck Group (1 mentions) Protein g dynabead, by Thermo Fisher Scientific (1 mentions) H3k9me3, by Abcam (1 mentions) Flag tag (flag), by Merck Group (1 mentions) Bioruptor pico, by Diagenode (1 mentions) Ab12286, by Abcam (1 mentions) Sc 20703, by Santa Cruz Biotechnology (1 mentions) Ab1791, by Abcam (1 mentions) Na934v, by GE Healthcare (1 mentions) Na931v, by GE Healthcare (1 mentions) Anti tubulin, by Merck Group (1 mentions) T6199, by Merck Group (1 mentions) F3165, by Merck Group (1 mentions) Ab191698, by Abcam (1 mentions) Sox2 sc 17320, by Santa Cruz Biotechnology (1 mentions) Bca assay kit, by Thermo Fisher Scientific (1 mentions)

3 protocols using abe364

Chromatin Immunoprecipitation of SMC Contractile Genes

Cited 1 time

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Chromatin Immunoprecipitation (ChIP) was performed as previously described (Dahl and Collas, 2008 (link); Gomez et al., 2013 (link)). In brief, passage-matched control, Myocd-LSD1, Myocd-LSD1^NF SMC were fixed with 1% PFA for 10 min at room temperature. Cells were sonicated with a Bioruptor Pico (Diagenode) to obtain chromatin fragments of 200-500 base pairs. Chromatin was incubated with Protein G Dynabeads (Invitrogen, 10004D) and one of the following antibodies: H3K4me2 (2μg; #07-030, Millipore), Flag (4;g #F3165, Sigma Aldrich), H3K9me3 (2μg; ab176916, Abcam), H3K27me3 (2μg; 07-449, Sigma Aldrich), SRF (2μg; sc-13029, Santa Cruz), KLF4 (2μg; sc-20691, Santa Cruz), TET2 (5μg; ABE364, Millipore), H3ac (2μg; 06-599, Millipore), rabbit IgG (Abcam, ab171870) or mouse IgG (Abcam, ab37355). Genomic DNA was extracted with phenol-chloroform from immunoprecipitated (IP) and non-immunoprecipitated (INPUT) samples. Histone modification and protein enrichment was measured by qPCR using primer sets targeting CArG regions of the SMC contractile genes. Results were expressed as IP/INPUT. Primers used for ChIP-qPCR are listed in Key Resources Table.

Liu M., Espinosa-Diez C., Mahan S., Du M., Nguyen A.T., Hahn S., Chakraborty R., Straub A.C., Martin K.A., Owens G.K, & Gomez D. (2021). H3K4 di-methylation governs smooth muscle lineage identity and promotes vascular homeostasis by restraining plasticity. Developmental cell, 56(19), 2765-2782.e10.

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Comprehensive Western Blot Antibody Panel

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For western blot, the following antibodies were used at the dilutions indicated:
Anti-FLAG (1:3000, F3165, Sigma-Aldrich); Anti-myc (1:3000, 2276, CST); Anti-HA (1:750, 3724, CST); Anti-Tubulin (1:20000, T6199, Sigma-Aldrich); Anti-Drosha (1:2000, 3364, CST; 1,2000, SC393591, Santa Cruz Biotechnology; 1:2000, ab12286, abcam); Anti-Dnmt1 (1:2000, 5032, CST); Anti-Dnmt3A (1:3000, SC20703, Santa Cruz Biotechnology); Anti-Dnmt3B (1:3000, SC52922, Santa Cruz Biotechnology); Anti-Dnmt3L (1:1000, 12309, CST); Anti-Uhrf1 (1:800, SC98817, Santa Cruz Biotechnology); Anti-Histone H3 (1:2000, ab1791, abcam); Anti-Tet1 (1:1000, 09–872, Millipore); Anti-Tet2 (1:1000, ABE364, Millipore); Donkey anti-rabbit HRP (1:5000, NA934V, GE Healthcare); Sheep anti-mouse HRP (1:5000, NA931V, GE Healthcare).

Stathopoulou A., Chhetri J.B., Ambrose J.C., Estève P.O., Ji L., Erdjument-Bromage H., Zhang G., Neubert T.A., Pradhan S., Herrero J., Schmitz R.J, & Ooi S.K. (2017). A novel requirement for DROSHA in maintenance of mammalian CG methylation. Nucleic Acids Research, 45(16), 9398-9412.

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Pluripotency Protein Expression Analysis

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Cells were lysed in SDS buffer. The protein concentration was measured by BCA assay kit (Thermo). Equal amounts of cell lysates were loaded, blotted onto a polyvinylidene difluoride (PVDF) membrane, and probed with the following primary antibodies: Oct4 (SC-8628, 1:1000, Santa Cruz), Sox2 (SC-17320, 1:1000, Santa Cruz), Tet1 (ab191698, 1:500, Abcam, Cambridge, UK), Tet2 (ABE364, 1: 1000, Millipore), and GAPDH (ab8245, 1:4000, Abcam). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the loading control. After incubation with the appropriate secondary antibodies, signals were visualized by enhanced chemiluminescence (GE systems, Fairfield, USA).

Tu J., Tian G., Cheung H.H., Wei W, & Lee T.L. (2018). Gas5 is an essential lncRNA regulator for self-renewal and pluripotency of mouse embryonic stem cells and induced pluripotent stem cells. Stem Cell Research & Therapy, 9, 71.

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