Immunofluorescence staining was performed as previously described (Chen et al., 2020 (link)). The primary antibodies included mouse anti-CC1 (1:200, Millipore, Cat: OP80), rabbit anti-NG2 (1:200, Millipore, Cat: MAB5320), mouse anti-Olig2 (1:300, Millipore, Cat: MABN50), rabbit anti-Olig2 (1:300, Millipore, Cat: AB9610), rabbit anti-Sox10 (1:200, Abcam, Cat: ab180862), mouse anti-5mC (1:500, Abcam, Cat: ab10805), and rabbit anti-Ki67 (1:500, Thermo Fisher Scientific, Cat: MA514520). For 5mC staining, before being blocked in 5% BSA, the brain slices were treated with 70% ethanol (pre-cooled on ice) for 5 min and then treated with 1.5 M HCl for 30 min at room temperature.
Mouse anti 5mc
Manufactured by Abcam
Sourced in United Kingdom
Mouse anti-5mC is an antibody that specifically recognizes 5-methylcytosine (5mC), a modified form of the DNA base cytosine. This antibody can be used to detect and study DNA methylation, a crucial epigenetic modification involved in gene regulation and cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti ki67, by Thermo Fisher Scientific (1 mentions)
Rabbit anti olig2, by Merck Group (1 mentions)
Mouse anti cc1, by Merck Group (1 mentions)
Rabbit anti ng2, by Merck Group (1 mentions)
Rabbit anti sox10, by Abcam (1 mentions)
Mouse anti olig2, by Merck Group (1 mentions)
Goat anti rabbit hrp, by Cell Signaling Technology (1 mentions)
Goat anti mouse hrp, by Cell Signaling Technology (1 mentions)
Sonication, by Diagenode (1 mentions)
Magnetic beads, by Active Motif (1 mentions)
Rabbit anti mecp2, by Cell Signaling Technology (1 mentions)
Rabbit anti 5hmc, by Active Motif (1 mentions)
Rabbit anti nurr1, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti tet1, by Abcam (1 mentions)
3 protocols using mouse anti 5mc
1
Immunofluorescence Staining of Neural Cell Markers
2
Immunofluorescence Staining of Folate Receptor
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Primary antibodies used were anti-rabbit ALDH1L1 at 1:2000 dilution and rabbit anti-FRĪ± at 1:10,000 (Proteintech, Manchester, UK), mouse anti-folic acid (Sigma Aldrich, Gillingha, UK) at 1:2000, mouse anti-5mc at 1:500, and mouse anti-5hmc (Abcam, Cambridge, UK), rabbit anti-DNMT1 (Epigentek, Farmingdale, NY, USA) at 1:500, mouse anti-beta-actin (Cell Signalling Techology, Leiden, The Netherlands) at 1:10,000. Secondary antibodies were goat anti-rabbit HRP and goat anti-mouse HRP (Cell Signalling Technology, Leiden, The Netherlands ), or goat anti-rabbit Coralite 488 or 594 and goat anti-mouse Coralite 488 or 594 (all from Proteintech, Manchester, UK). Alternatively, Alexa Fluor-conjugated secondary antibodies were also used at 555, 488, Cy7 (Abcam, Cambridge, UK).
3
Chromatin Immunoprecipitation and Methylation Analysis
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For ChIP experiments, cells were cross-linked with 1% paraformaldehyde for 15 minutes and Chromatins were sheared into an average 200-400 bp in length by sonication (Diagenode) and immunoprecipitated with following antibodies: rabbit anti-MeCP2 (Cell Signaling Technology), rabbit anti-NURR1 (Santa Cruz Biotechnology) and rabbit anti-TET1 (Abcam). Immunoprecipitated DNA fragments were collected by magnetic beads (Active Motif), purified, and subjected to real-time PCR using primers specific to regions spanning three NBREs on Th promoter. Data were normalized to values of the input DNA. For MeDIP and hMeDIP experiments, genomic DNA were extracted from cells, sheared, immunoprecipitated, collected and subjected to real-time PCR in a similar way as ChIP with following antibodies: mouse anti-5mC (Abcam) and rabbit anti-5hmC (Active Motif). The primers are listed in Supplemental Table 1.
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