Rat lungs were homogenized, and cells were lysed in lysis buffer (50 mmol/L Tris–HCl pH 7.4, 150 mmol/l NaCl, 0.5% nonidet P-40) containing EDTA-free complete protease and phosphatase inhibitor (Sigma–Aldrich/Roche). Protein levels were measured using Bradford assays (Bio-Rad), after which 35 μg samples were run on SDS-PAGE gels, transferred to nitrocellulose membranes, blocked with either 5% bovine serum albumin or 4% milk, and subsequently incubated with primary and secondary antibodies and detected using SuperSignal West Pico Chemiluminescent Substrate (catalog no.: # 34080, Thermo Fisher Scientific) on iBright. Protein levels were determined through densitometric analysis using ImageJ software (NIH). The following previously validated primary antibodies were used: DNMT1 (Cell Signaling), DNMT3A (Cell Signaling), DNMT3B (Epigentek), MAT2A (Novus Biologicals), SAHH (Santacruz), NOS3 (Novus Biologicals), SOD2 (Novus Biologicals), SERPINE1 (Novus Biologicals), and ABRA (Novus biologicals). ACTB (Cell Signaling) was used as a loading control.
Dnmt3b
Manufactured by Epigentek
Sourced in United States
DNMT3B is a DNA methyltransferase enzyme that catalyzes the addition of methyl groups to cytosine residues in DNA, playing a crucial role in de novo DNA methylation.
Automatically generated - may contain errors
Lab products found in correlation
Mat2a, by Novus Biologicals (1 mentions)
Dnmt3a, by Cell Signaling Technology (1 mentions)
Dnmt1, by Cell Signaling Technology (1 mentions)
Bradford assay, by Bio-Rad (1 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Electrophoresis reagents, by Bio-Rad (1 mentions)
Dnmt1, by Epigentek (1 mentions)
Cell culture reagents, by ScienCell (1 mentions)
Dnmt3a, by Epigentek (1 mentions)
Mitochondrial isolation kit, by Abcam (1 mentions)
2 protocols using dnmt3b
1
Protein Expression Analysis in Rat Lungs
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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