Quantitative Real-time PCR (TaqMan®; Applied Biosystems; Foster City, CA, USA) was performed using i) specific primers and probes for peroxisome-proliferator activated receptor-gamma co-activator 1 alpha (Ppargc1a; NM_031,347, Fw Primer 5′-CGATCACCATATTCCAGGTCAAG-3’; Rv primer 5′-CGATGTGTGCGGTGTCTGTAGT-3’; Probe FAM-5′-AGGTCCCCAGGCAGTAGATCCTCTTCAAGA-3′-TAMRA) or ii) commercially available validated TaqMan® primers and probes (Applied Biosystems; Foster City, CA, USA) for peroxisome-proliferator activated receptor-gamma co-activator 1 beta (PGC1β, Ppargc1b; Assay ID Rn_00598,552_m1), peroxisome-proliferator-activated receptor gamma (PPARγ, Pparg; Assay ID Rn_00440,945_m1), PR domain containing 16 (PRDM16, Prdm16; Assay ID Rn_01516224_m1), cell death-inducing DFFA-like effector a (Cidea, Cidea; Assay ID Rn_04181355_m1), otopetrin 1 (OTOP1, Otop1; Assay ID Rn_00710,679) and iodothyronine deiodinase 2 (DIO2, Dio2; Assay ID Rn_00581,867_m1) [27 ,32 (link),34 (link)]. Values were expressed in relation to hypoxanthine-guanine phosphoribosyl-transferase levels (Hprt; NM_012,583, FW Primer 5′-AGCCGACCGGTTCTGTCAT-3’; Fw Primer 5′-GGTCATAACCTGGTTCATCATCAC-3’; Probe FAM-5′-CGACCCTCAGTCCCAGCGTCGTGAT-3′-TAMRA).
Cidea
Manufactured by Thermo Fisher Scientific
Sourced in United States
Cidea is a laboratory equipment product offered by Thermo Fisher Scientific. It is a device used for the measurement and analysis of various biological and chemical samples. The core function of Cidea is to provide accurate and reliable data for research and testing purposes.
Automatically generated - may contain errors
Lab products found in correlation
Ppargc1a, by Thermo Fisher Scientific (2 mentions)
Taqman primers and probe, by Thermo Fisher Scientific (1 mentions)
Gata3, by Thermo Fisher Scientific (1 mentions)
Prdm16, by Thermo Fisher Scientific (1 mentions)
Rneasy lipid tissue mini kit, by Qiagen (1 mentions)
Random hexamer, by Cytiva (1 mentions)
Prism 7900 system, by Thermo Fisher Scientific (1 mentions)
3 protocols using cidea
1
Quantitative Real-time PCR of Metabolic Regulators
2
Quantification of Gene Expression in Tissues
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Total RNA from cells was extracted with TRIzol reagent. For RNA analysis from blood, mice were cheek bled into a mixture of sodium citrate (final 2%) and RPMI. The cell pellet was incubated with ammonium-chloride-potassium (ACK) lysis buffer. The remaining pellet was suspended in TRIzol for RNA extraction. Total RNA from tissue was isolated by homogenization with an Omni Tissue Homogenizer in QIAzol Lysis Reagent using an RNeasy Lipid Tissue Mini Kit (Qiagen 74804). RNA was converted to cDNA using ProtoScript II Reverse Transcriptase. Predesigned TaqMan gene expression assays were purchased from Applied Biosystems: Klk1b22 (Mm02343755_g1, Mm02343756_g1, Mm02343754_m1), uncoupling protein 1 (Ucp1) (Mm01244861_m1), Cidea (Mm00432554_m1), Cpt1b (Mm01308160_m1), Prdm16 (Mm00712556_m1), Tbx21 (Mm00450960_m1), Gata3 (Mm00484683_m1), Rorc (Mm01261022_m1), Ifng (Mm01168134_m1), and Ppargc1a (Pgc1α) (Mm01208835_m1). Quantitative RT-PCR (qRT-PCR) was performed with EagleTaq Universal Master Mix using an Applied Biosystems StepOnePlus 96-well RT-PCR. ΔΔCt values were normalized to 18S rRNA (Applied Biosystems 4310893E) and to a control group of interest or background.
3
Gene Expression Analysis of Metabolic Regulators
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Total RNA was isolated from the tissue using Qiazol lysis reagent (Quiagen Sciences, USA). Two μg of RNA were treated with DNase I (DNase I amplification grade; Invitrogen, CA) and then reversely transcribed using random hexamers (Amersham Biosciences, NJ) and SuperScript II reverse transcriptase (Invitrogen, CA). Quantitative real-time polymerase chain reactions were performed using ABI PRISM 7900 System with TaqMan primers bmp7 (Mm00432102_m1), ppargc1a (Mm01208835_m1), fgf21 (Mm00840165_g1), elovl3 (Mm00468164_m1), rps29 (Mm02342448_gH), tbp (Mm01277042_m1), cidea (Mm00432554_m1), car4 (Mm00483021_m1), tfam (Mm00447485_m1), nrf1 (Mm01135606_m1) and reagents (Applied Biosystems, CA); Qiagen SYBR green primers were Prdm16 (QT00148127), tbx15 (QT00148127), zic1 (QT00173502), tcf21 (QT00100688), ucp1 (QT00097300), cpt1 (QT00172564) and tbp (QT00198443). Each reaction was performed in duplicate and results were normalized to the geometric average of two internal controls (Rps29 and Tbp).
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