CDCA and CA were purchased from Sigma-Aldrich. FXR agonist GW4064 was obtained from Tocris Biosciences. Cell culture reagents Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), TaqMan master-mix and probes were purchased from Life Technologies. RNA Bee for RNA isolation, dimethyl sulfoxide (DMSO), and propanediol were obtained from Fisher Scientific. Primary and secondary antibodies for Western blotting were purchased from Santa Cruz Biotechnologies. All Western blotting gels, buffers, and markers were purchased through BioRad Laboratories. Complementary DNA synthesis kit was purchased through Promega. Protease inhibitor, Halt, and bovine serum albumin quantification reagents were purchased from Thermo Scientific.
Complementary dna synthesis kit
Manufactured by Promega
The Complementary DNA synthesis kit is a laboratory tool used to convert RNA into complementary DNA (cDNA) molecules. This process, known as reverse transcription, allows for the study of gene expression and analysis of RNA samples.
Automatically generated - may contain errors
Lab products found in correlation
Nanodrop 1000, by Thermo Fisher Scientific (2 mentions)
Protease inhibitor, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Gw4064, by Bio-Techne (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Taqman master mix, by Thermo Fisher Scientific (1 mentions)
Rotor gene 6000 system, by Qiagen (1 mentions)
2 protocols using complementary dna synthesis kit
1
Investigating FXR Agonist-Mediated Responses
2
Quantifying PAFr mRNA Expression in Airway Cells
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Cells were seeded into sterile clear-flat bottom 24-well plates (Corning Inc) before stimulation with CSE (1%, 4 hours). RNA was prepared using the Reliaprep™ Mini RNA cell Miniprep system (Promega, Sydney, Australia) with total RNA quantified by Nanodrop 1000 (Thermo Fisher Scientific, Waltham, MA, USA). Complementary DNA was then collected using the Promega complementary DNA synthesis kit. The level of PAFr transcript was determined by quantitative polymerase chain reaction using the Corbett Rotor-Gene 6000 system (Qiagen, Hilden, Germany). Thermocycling controls were run as previously described.25 (link) The relative change of PAFr mRNA expression was normalized to three reference genes (18S rRNA, β-actin, and β2-microglobulin) using the primers and recommended conditions provided by the manufacturer (Qiagen, Catalogue # PPH05820C). Data were derived from two independent experiments performed in duplicate.
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