Human Endothelial Culture Medium was purchased from Thermo Scientific (Grand Island, NY, USA). The FITC-Annexin V/PI Apoptosis Detection Kit was obtained from BD Biosciences (Franklin Lakes, NJ, USA). The Promoter-Binding TF Profiling Plate Array I (FA-2001) was obtained from Signosis (Santa Clara, CA, USA). Dual-Luciferase Reporter Assay System (E1960), PGL4.12 (E6671) and PGL4.73 (E6911) vectors were purchased from Promega (Madison, WI, USA). The ChIP assay kit was obtained from Millipore (Billerica, MA, USA). Plasmid pcDNA3.1 (+), a terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labelling (TUNEL) kit and TRIzol Reagent were purchased from Invitrogen (Grand Island, NY, USA). Antibodies against CREG (ab191909, ab68341) and GATA1 (ab181544) were purchased from Abcam (Cambridge, MA, USA). Antibodies against cleaved caspase-3 and GAPDH were obtained from Cell Signalling Technology (Danvers, MA, USA). The enhanced chemiluminescence (ECL) Western Blotting System was purchased from GE Life Sciences (Marlborough, MA, USA). D-glucose, free fatty acid (FFA)-free bovine serum albumin (BSA), palmitate and other chemicals, if not specified otherwise, were purchased from Sigma-Aldrich (St Louis, MO, USA).
Promoter binding tf profiling plate array 1
Manufactured by Signosis
Sourced in United States
The Promoter-Binding TF Profiling Plate Array is a laboratory tool designed for the simultaneous detection and evaluation of transcription factor binding to specific DNA promoter sequences. It provides a platform for high-throughput analysis of transcription factor-DNA interactions.
Automatically generated - may contain errors
Lab products found in correlation
Ab181544, by Abcam (1 mentions)
Antibodies against cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Ecl western blotting system, by GE Healthcare (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by BD (1 mentions)
Chip assay kit, by Merck Group (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
D glucose, by Merck Group (1 mentions)
2 protocols using promoter binding tf profiling plate array 1
1
Endothelial Cell Culture and Apoptosis Analysis
2
Transcription Factor Binding Assay
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Assay was performed as recommended by the manufacturer of Promoter-Binding TF Profiling Plate Array I (Signosis). Briefly, nuclear extract was isolated from 5 9 10 6 HD11 cells, and the protein content was estimated. The reaction mixture, including 15 ll of TF binding buffer, 3 ll of probes, 10 lg of nuclear extract and 4 ll of the NLRC5 promoter fragment or no DNA, was incubated at room temperature for 30 min to allow for formation of the TF-DNA complex. Unbound probes were separated from the complex, and bound probes were eluted and then hybridized to the plate and incubated overnight at 42 °C. Bound probes were detected using a HRP-streptavidin conjugate incubated with the chemiluminescent substrate. Finally, chemiluminescence was measured.
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