30 μg of cell lysates were analyzed on 8% Tris-Glycine polyacrylamide gels (Invitrogen) and transferred to PVDF membrane using BioRad mini transfer apparatus for 1.5 hour at 50 V. Membrane was blocked with 5% milk for one hour in room temperature and probed with Rabbit polyclonal anti-DNMT1 (Abcam, 1:1000), DNMT3A (Abcam, 1:500), DNMT3B (Abcam, 1:500), β-actin (Abcam, 1:500), Tet1 antibody (Abcam, 1:1000) or mouse monoclonal anti-TET2 (Active Motif 1:500) for overnight at 4°C. This was followed by washes in PBST (0.5% Tween 20) and incubation in HRP-conjugated secondary antibodies (Cal Biochem). Blots were developed using ECL reagent (GE Healthcare).
Tet1 antibody
Manufactured by Abcam
Tet1 antibody is a laboratory reagent used to detect the presence of the Tet1 (Ten-Eleven Translocation 1) protein in biological samples. Tet1 is an enzyme involved in DNA demethylation, which plays a role in regulating gene expression. This antibody can be used in various techniques, such as Western blotting, immunohistochemistry, and immunocytochemistry, to analyze the expression and localization of Tet1 in cells and tissues.
Automatically generated - may contain errors
2 protocols using tet1 antibody
1
Western Blot Analysis of DNA Methylation Enzymes
2
miRNA-21-5p Inhibition Suppresses T98G Xenograft Tumor Growth
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BALB/c nude mice (n = 12) were purchased from Shanghai Slake Laboratory Animal Co., Ltd (Shanghai, China). 100 μl of T98G cells (2 × 10 6 ) was subcutaneously inoculated into nude mice. The tumour sizes were recorded with vernier callipers, and the volume was calculated using the following formula: Tumour volume = 0.5 × length × width 2 . When the average tumour volume reached approximately 100-150 mm 3 , the mice were divided into 2 groups (antagomir-NC group, n = 6; antagomir-21-5p group, n = 6). miRNA antagomirs (antagomirs) are synthetic antagonists of miRNAs as silencing agents of miRNAs [19] . 20 nmol antagomir-21-5p or antagomir-NC was injected intratumorally every 3 days. After 4 weeks, nude mice were euthanized, and the xenograft tumour was excised. Tumour tissues were collected and subjected to immunohistochemical staining using TET1 antibody (1 : 200, Abcam). This study was approved by the Ethics Committee (No. 20191009) of Liaocheng Second People's Hospital.
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