The recombinant adenoviruses expressing the DNMT1 (Adv-DNMT1) gene were constructed with a replication-defective adenoviral shuttle vector pAdtrack–cytomegalovirus-green fluorescent protein as described (32 (link)). Western blotting was done to examine ectopic gene expression using antibodies against DNMT1 (60B1220.1, IMG-261A; IMGENEX, San Diego, CA). The recombinant adenovirus encoding only the green fluorescent protein was used as a control (CT).
Img 261a
Manufactured by Novus Biologicals
Sourced in United States
The IMG-261A is a compact and versatile imaging system designed for a wide range of applications in life science research. It provides high-quality imaging capabilities for various sample types, including gels, blots, and plates. The IMG-261A is capable of capturing images with excellent resolution and sensitivity, making it a valuable tool for researchers in the field of molecular biology and biochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Dnmt1, by Novus Biologicals (1 mentions)
Cebpd sc 636, by Santa Cruz Biotechnology (1 mentions)
Casp8, by Thermo Fisher Scientific (1 mentions)
α tubulin, by Merck Group (1 mentions)
Dnmt3a, by Novus Biologicals (1 mentions)
Ar pg 21, by Merck Group (1 mentions)
Dnmt3b, by Novus Biologicals (1 mentions)
Casp3, by Cell Signaling Technology (1 mentions)
3 protocols using img 261a
1
Adenoviral Expression of DNMT1
2
Protein Expression and Apoptosis Assay
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The experimental cells were harvested and lysed in RIPA buffer (50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1 mM EDTA, 1% Nonidet P-40, 0.25% sodium deoxycholate, 1 mM DTT, 10 mM sodium fluoride, 1 mM phenylmethylsulfonyl fluoride, aprotinin 1 μg/ml, leupeptin 1 μg/ml and 1 mM Na3VO4). The proteins were separated by SDS-PAGE, transferred to PVDF membranes and probed with specific antibodies as follows: AR (pg-21, Millipore, Billerica, MA, USA), CEBPD (SC-636, Santa Cruz, Dallas, TX, USA), DNMT1 (IMG-261A, IMGENEX, San Diego, CA, USA), DNMT3A (IMG-268A, IMGENEX), DNMT3B (IMG-184A, IMGENEX), E2F1 (SC-193, Santa Cruz), EZH2 (07-400, Upstate, Billerica, MA, USA), HA (MMS-101R, COVACE, Princeton, NJ, USA), H3K27 trimethylation, (07-449, Upstate), myc (SC-40, Santa Cruz), CASP8 (RB-1200, Thermo Scientific, Pittsburgh, PA, USA), CASP3 (9661, Cell Signaling Technology, Danvers, MA, USA), SUZ12 (07-379, Upstate), tBid (2002, Cell Signaling Technology) and α-tubulin (T6199, Sigma). Mitochondrial protein and cytosolic protein were isolated using the Cytochrome c Releasing Apoptosis Assay kit according to the manufacturer's instructions (K257-100, BioVison, San Francisco, CA, USA).
3
Ubiquitination of DNMT1 in HEK293 Cells
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HEK293 cells expressing shUbe3a, shUbe3a + Ube3a cDNA, or Ube3a cDNA were treated with 10 µM MG132 (American Peptide, Sunnyvale, CA) for 90 min after 72‐h posttransfection. The cells were washed twice with PBS, lyzed by RIPA buffer, and centrifuged to extract the supernatant, which was precleared by the control IgG and protein A/G PLUS‐agarose. The cleared cell lysates containing 2 mg total protein was incubated with the DNMT1 primary antibody (IMG‐261A, IMGENEX) and protein A/G PLUS‐Agarose at 4°C overnight, washed with PBS, boiled and loaded in SDS‐PAGE and immunoblotted with the ubiquitin antibody (sc‐8017, Santa Cruz).
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