Rabbit anti ctcf

Manufactured by Merck Group

Rabbit anti-CTCF is a primary antibody that recognizes the CTCF (CCCTC-binding factor) protein. CTCF is a highly conserved and ubiquitously expressed zinc finger protein that plays a critical role in gene regulation, chromatin organization, and DNA methylation. This antibody can be used in various applications, such as Western blotting, immunohistochemistry, and chromatin immunoprecipitation (ChIP), to detect and study the CTCF protein.

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2 protocols using rabbit anti ctcf

Nuclear Protein Extraction and Analysis

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The nuclei (weight, 50 µg) were incubated on ice for 15 min in a series of buffers: HE (0 mM Mg²⁺ buffer), H10Mg1, and H10Mg5. After incubation, centrifugation was performed to recover nuclei. The nuclear pellets were suspended in a final sample buffer (Laemmli, 1970 (link)) and subjected to 12.5% SDS–PAGE. Subsequently, the gels were subjected to Coomassie brilliant blue (CBB) staining and Western blotting using the following antibodies: mouse anti-Rad21 (05-908; Millipore), rabbit anti-CTCF (07-729; Millipore), rabbit anti-Smc2 (ab10412; Abcam), rabbit anti-H2B (ab1790; Abcam), rabbit anti–acetyl histone H4 (06-866; Millipore), rabbit anti–acetyl histone H3 (06-599; Millipore), and goat anti–lamin A/C (sc-6215; Santa Cruz Biotechnology).

Shimamoto Y., Tamura S., Masumoto H, & Maeshima K. (2017). Nucleosome–nucleosome interactions via histone tails and linker DNA regulate nuclear rigidity. Molecular Biology of the Cell, 28(11), 1580-1589.

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Profiling Histone Modifications and Transcription Factors via AutoCUT&RUN

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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.

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