Rabbit anti dnmt1

Manufactured by Cell Signaling Technology

Sourced in United States

Rabbit anti-DNMT1 is a primary antibody that recognizes the DNA methyltransferase 1 (DNMT1) protein. DNMT1 is responsible for maintaining DNA methylation patterns during DNA replication and repair.

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DNMT1 (67 citations) Anti-DNMT1 (32 citations)

5 protocols using rabbit anti dnmt1

Protein Expression Analysis in DRG Neurons

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As previously described ^{30 (link), 31}, bilateral lumber mice DRGs were collected. DRG tissues were homogenized and centrifugalized at 4 °C. The supernatant was collected, and the pellet (nuclei and debris fraction) was dissolved in lysis buffer and then sonicated on ice. The membrane or nuclei samples were heated at 99°C for 5 min, loaded and run onto precast polyacrylamide gel (Bio-Rad), then were electrophoretically transferred onto nitrocellulose membranes. After blocking, The membranes were incubated with primary rabbit anti-K_2p1.1 (1:1,000, Alomone), mouse anti-K_v1.2 (1:500, NeuroMab), rabbit anti-GAPDH (1:2,000, Santa Cruz Biotechnology), rabbit anti-DNMT3a (1:500, Cell Signaling Technology), rabbit anti-DNMT3b (1:500, Santa Cruz Biotechnology), rabbit anti-DNMT1 (1:1,000, Cell Signaling Technology), or rabbit anti-H3 (1:2,000, Cell Signaling Technology) at 4 °C overnight. After secondary antibody incubation, the blots were developed with chemiluminescent regents. The intensities of blots were quantified with Image Lab software.

Mao Q., Wu S., Gu X., Du S., Mo K., Sun L., Cao J., Bekker A., Chen L, & Tao Y.X. (2019). DNMT3a-triggered downregulation of K2p1.1 gene in primary sensory neurons contributes to paclitaxel-induced neuropathic pain. International journal of cancer, 145(8), 2122-2134.

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Protein Expression Analysis of Stem Cells

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Cells were lysed using lysis buffer and extracted for 20 min on ice. Whole-cell extractprotein (30 μg) was subjected to electrophoresis on 10% SDS-PAGE gel and transferred to nitrocellulose membranes. Then membranes were incubated with the following primary antibodies: rabbit anti-Nanog, anti-Oct-4, anti-Sox-2 (dilution 1:1000, Proteintech, Rosemont, USA), rabbit anti-DNMT1, anti-DNMT3A, anti-DNMT3B (dilution 1:1000, Cell Signaling Technology, Danvers, MA, USA), and rabbit anti-GAPDH (dilution 1:2000, Antgene, Wuhan, China).

Ai L., Mu S., Sun C., Fan F., Yan H., Qin Y., Cui G., Wang Y., Guo T., Mei H., Wang H, & Hu Y. (2019). Myeloid-derived suppressor cells endow stem-like qualities to multiple myeloma cells by inducing piRNA-823 expression and DNMT3B activation. Molecular Cancer, 18, 88.

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Immunohistochemical Analysis of Mouse Embryos

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Mouse embryos and fetuses at indicated time points were fixed in 4% paraformaldehyde overnight and dehydrated through a graded series into 100% methanol for storage at −20 °C. Embryos and fetuses were rehydrated, and immunohistochemistry was performed as previously described (73 (link)). The following primary and secondary antibodies were used: 1:50 rabbit anti-DNMT1 (#5032, Cell Signaling Technologies), 1:200 mouse anti-CDH1 (BD 610181), 1:250 DyLight 488-conjugated goat anti-mouse IgG, and 1:250 DyLight 594-conjugated goat anti-rabbit IgG (#35502 and #35560, Jackson ImmunoResearch). Sections were imaged using a Keyence BZ-X700 (Keyence) fluorescence microscope.

Ulschmid C.M., Sun M.R., Jabbarpour C.R., Steward A.C., Rivera-González K.S., Cao J., Martin A.A., Barnes M., Wicklund L., Madrid A., Papale L.A., Joseph D.B., Vezina C.M., Alisch R.S, & Lipinski R.J. (2024). Disruption of DNA methylation–mediated cranial neural crest proliferation and differentiation causes orofacial clefts in mice. Proceedings of the National Academy of Sciences of the United States of America, 121(3), e2317668121.

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Western Blotting Assay for Protein Expression

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INS‐1 cells were harvested with ice‐cold lysis buffer supplemented with complete proteinase inhibitor mixture (Roche). Other specific information of Western blotting assay was performed as previously described previously.¹⁹ The antibodies used are as follows: rabbit anti‐CASK antibody (1:1000; Cell Signaling), rabbit anti‐DNMT1 (1:1000; Cell Signaling), mouse anti‐DNMT3a (1:1000; Santa Cruz Biotechnology), rabbit anti‐DNMT3b (1:1000; Abcam), rabbit anti‐Akt(pan) antibody (1:1000; Cell Signaling), rabbit anti‐p‐Akt Ser473 antibody (1:5000; Abcam), rabbit anti‐iNOS Antibody (1:1000; Cell Signaling) or rabbit anti‐β‐tubulin antibody (1:4000; Cell Signaling).

Wang T., Liu X., Xie J., Yuan Q, & Wang Y. (2020). Cask methylation involved in the injury of insulin secretion function caused by interleukin1‐β. Journal of Cellular and Molecular Medicine, 24(24), 14247-14256.

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Investigating PTEN Regulation and Epigenetics

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Bpv (PTEN) and 5-Aza-2 0 -deoxycytidine were obtained from Sigma Inc. (St. Louis, MO, USA). Complete Freund's adjuvant was purchased from Chondrex, Inc. (WA, USA). Rabbit anti-PTEN and anti-TNF-a monoclonal antibody were obtained from Abcam (Cambridge, UK). Rabbit anti-DNMT1 and anti-Vimentin (Alexa Fluor 594 Conjugated) were acquired from Cell Signaling Technology (Danvers, MA, USA). Rabbit anti-IL-6 and anti-b-actin monoclonal antibody were purchased from Bioworld (Shanghai, China). Peroxidase-conjugated goat antirabbit IgG (H þ L) was obtained from ZSGB-BIO (Beijing, China). PTEN, TNF-a, IL-6, IL-1b, CCL-2, CCL-3, VCAM-1, and VEGF-a and b-actin primers were obtained from Sangon Biotech (Shanghai) Co., Ltd (Shanghai, China).

Li X.F., Chen X., Bao J., Xu L., Zhang L., Huang C., Meng X.M, & Li J. (2019). PTEN negatively regulates the expression of pro-inflammatory cytokines and chemokines of fibroblast-like synoviocytes in adjuvant-induced arthritis. Artificial cells, nanomedicine, and biotechnology, 47(1).

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