Anti rnapol 2

Manufactured by Cell Signaling Technology

Anti-RNApol II is a product from Cell Signaling Technology that detects RNA polymerase II. It is a primary antibody used for Western blotting, immunoprecipitation, and immunohistochemistry applications.

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Lab products found in correlation

Anti h3k18ac, by Active Motif (2 mentions) Lightcycler, by Roche (2 mentions) Ab992, by Abcam (1 mentions) Ab4729, by Abcam (1 mentions) Anti histone h3, by Merck Group (1 mentions) Chip assay kit, by Merck Group (1 mentions) Anti h3k27me3, by Merck Group (1 mentions) Sensifast sybr hi rox kit, by Meridian Bioscience (1 mentions) Anti pr h 190 sc 7208, by Santa Cruz Biotechnology (1 mentions) Anti ctcf, by Merck Group (1 mentions) Anti h3k27ac, by Abcam (1 mentions)

3 protocols using anti rnapol 2

Quantitative Chromatin Immunoprecipitation Analysis

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Chromatin immunoprecipitation (ChIP) assays were performed as described (12 (link)) using anti-PR (H190 SC-7208, Santa Cruz), anti-ERK2(D12) (05-157, Merck), anti-SRC3-(5E11) (#2126, Cell Signaling), anti-p300, clone NM11 (#61401, Active Motif), anti-RNApol II (#2629, Cell Signaling), anti-RAD21 (ab992, Abcam), anti-CTCF (07-729, Merck), anti-H3K27ac (ab4729, Abcam), anti-H3K18ac (#39693, Active Motif) and anti-BRD4 (kind gift from Cheng-Ming Chiang, UT Southwestern Medical Center). Quantification of ChIP was performed by real-time PCR using Roche LightCycler (Roche). The fold enrichment of target sequence in the immunoprecipitated (IP) compared to input (Ref) fractions was calculated using the comparative Ct (the number of cycles required to reach a threshold concentration) method with the equation 2^{Ct(IP) − Ct(Ref)}. Each of these values was corrected by the human β-globin gene and referred to as relative abundance over time zero. Primer sequences for HAs, medium accessible sites (MAs) and low accessible sites (LAs) are available in Supplementary Table S1.

Zaurin R., Ferrari R., Nacht A.S., Carbonell J., Le Dily F., Font-Mateu J., de Llobet Cucalon L.I., Vidal E., Lioutas A., Beato M, & Vicent G.P. (2021). A set of accessible enhancers enables the initial response of breast cancer cells to physiological progestin concentrations. Nucleic Acids Research, 49(22), 12716-12731.

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ChIP Assay for Transcriptional Regulation

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ChIP assays were performed as described previously.⁶⁴ (link) Briefly, human monocytes were treated with GM-CSF (20 ng/ml) alone or together with Dex (100 nM) for 1 h before crosslinking protein-DNA complexes with 1% formaldehyde for 10 minutes at room temperature. ChIP was performed with a ChIP Assay Kit (17-295, Millipore) as per the manufacturer’s instructions. Immunoprecipitation was performed with 1μg of anti-RNA Pol II (#2629, Cell Signaling Technologies), anti-H3K27me3 (07-449, Millipore), anti-histone H3 (05-928, Millipore) antibodies, followed by reversal of cross-linking. Immunoprecipitated DNA fragments were then amplified by qPCR with a SensiFAST SYBR Hi-ROX Kit (Bioline) and the following specific primers for human IRF4 transcription start site (TSS) (forward 5′-ccacctcgcactctcagttt-3′ and reverse 5′-ctggaggtcgaacctctggt-3′). Enrichment of histones and RNA Pol II at the gene loci was expressed as percentage of input DNA.

Lupancu T.J., Lee K.M., Eivazitork M., Hor C., Fleetwood A.J., Cook A.D., Olshansky M., Turner S.J., de Steiger R., Lim K., Hamilton J.A, & Achuthan A.A. (2023). Epigenetic and transcriptional regulation of CCL17 production by glucocorticoids in arthritis. iScience, 26(10), 108079.

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Chromatin Immunoprecipitation (ChIP) Assay

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ChIP assays were performed as described (Strutt and Paro, 1999 (link)) using anti-PR (H190 SC-7208, Santa Cruz,); anti-RNApol II (#2629, Cell Signaling); anti-CTCF (07-729, Merck); anti-H3K27ac (ab4729, Abcam); anti-H3K18ac (#39693, Active Motif). Quantification of chromatin immunoprecipitation was performed by real-time PCR using Roche Lightcycler (Roche). The fold enrichment of target sequence in the immunoprecipitated (IP) compared to input (Ref) fractions was calculated using the comparative Ct (the number of cycles required to reach a threshold concentration) method with the Eq. (2) ^{Ct(IP)-Ct(Ref)}. Each of these values was corrected by the human β-globin gene and referred as relative abundance over time zero. Primers sequences for target regions are available on request.

Ramírez-Cuéllar J., Ferrari R., Sanz R.T., Valverde-Santiago M., García-García J., Nacht A.S., Castillo D., Le Dily F., Neguembor M.V., Malatesta M., Bonnin S., Marti-Renom M.A., Beato M, & Vicent G.P. (2024). LATS1 controls CTCF chromatin occupancy and hormonal response of 3D-grown breast cancer cells. The EMBO Journal, 43(9), 1770-1798.

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