Whole-cell extracts were prepared and protein concentration of samples determined using Bio-Rad DC Protein Assay (Bio-Rad, UK). Proteins were separated on 8% SDS polyacrylamide gel and transferred onto a nitrocellulose membrane in buffer containing 25 mM Tris, 192 mM glycine, and 20% methanol. In order to block nonspecific protein binding blots were incubated with 5% milk for 1h, followed by overnight incubation with primary antibody rabbit anti-TET2 (Abcam, ab135087), TET3 (Abcam, ab139805), DNMT3a (Abcam, ab2850), DNMT3b (Abcam, ab2851), DNMT1 (Epigentek, A-1001-100), TGFβ (Cell Signaling, (56E4) Rabbit mAb #3709) at 1:1000, rabbit anti-GAPDH (Abcam, ab9485) or mouse anti-β actin (Abcam, ab8226) both at 1:2000 dilution. Secondary antibody either against rabbit or mouse was used at 1:2000 dilution. The antibody complexes were detected by using Pierce ECL Western Blotting Substrate (Thermo Scientific).
Dnmt1
Manufactured by Epigentek
Sourced in United States
DNMT1 is a DNA methyltransferase enzyme that catalyzes the transfer of methyl groups to cytosine residues in DNA, thereby maintaining DNA methylation patterns during cell division. It plays a critical role in the epigenetic regulation of gene expression.
Automatically generated - may contain errors
Lab products found in correlation
Tgf β, by Cell Signaling Technology (1 mentions)
Dc protein assay, by Bio-Rad (1 mentions)
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Dnmt3b, by Epigentek (1 mentions)
Electrophoresis reagents, by Bio-Rad (1 mentions)
Cell culture reagents, by ScienCell (1 mentions)
Dnmt3a, by Epigentek (1 mentions)
Mitochondrial isolation kit, by Abcam (1 mentions)
2 protocols using dnmt1
1
Protein Expression and Epigenetic Regulation
2
Mitochondrial Isolation and Epigenetic Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cell culture reagents were purchased from ScienCell (Carlsbad, CA, USA). DNMT1, DNMT3a, DNMT3b, TET1, TET2, and TET3 antibodies; goat anti-rabbit IgG and goat anti-mouse IgG antibodies; and kits for analysis of global DNA methylation of 5mC were purchased from Epigentek Group Inc. (Farmingdale, NY, USA). Mitochondrial isolation kits were purchased from Abcam (CA). Electrophoresis reagents were purchased from Bio-Rad (Richmond, CA, USA), and nitrocellulose membranes were purchased from Amersham Scientific (Piscataway, NJ).
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