A portion of the WAT and BAT tissue was snap frozen in liquid nitrogen immediately after harvest. Tissue was disrupted using a Qiagen TissueLyser, followed by total RNA extraction using the Qiagen RNeasy Kit (Valencia, CA). cDNA was produced using ThermoScript RT-PCR Systems (Invitrogen, Carlsbad, CA). VEGF, Ucp1, and Dio2 transcript was analyzed using Taqman Gene Expression Assays: #Mm01281449; Mm01244861_m1, and Mm00515664_m1, respectively (Applied Biosystems). Transcript levels were determined relative to the signal from GAPDH (Taqman Gene Expression Assay, #Mm99999915_m1) and normalized to the mean value of samples from control mice.
Mm01244861 m1
Manufactured by Thermo Fisher Scientific
The Mm01244861_m1 is a qPCR primer and probe set designed for the detection and quantification of a specific gene target. It is intended for use in real-time reverse transcription PCR (RT-qPCR) applications.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (3 mentions)
Mm00515664 m1, by Thermo Fisher Scientific (2 mentions)
Mm00441242 m1, by Thermo Fisher Scientific (2 mentions)
Mm00443258 m1, by Thermo Fisher Scientific (2 mentions)
Stepone software program, by Thermo Fisher Scientific (2 mentions)
Steponeplus real time pcr system, by Thermo Fisher Scientific (2 mentions)
Thermoscript rt pcr system, by Thermo Fisher Scientific (1 mentions)
Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions)
Rneasy kit, by Qiagen (1 mentions)
Tissuelyser, by Qiagen (1 mentions)
Mn03302249 g1, by Thermo Fisher Scientific (1 mentions)
Mm01208835 m1, by Thermo Fisher Scientific (1 mentions)
Mm03302249 g1, by Thermo Fisher Scientific (1 mentions)
Mm00434228 m1, by Thermo Fisher Scientific (1 mentions)
Mm00840904 m1, by Thermo Fisher Scientific (1 mentions)
Mm99999915 g1, by Thermo Fisher Scientific (1 mentions)
7000 sequence detection system, by Thermo Fisher Scientific (1 mentions)
Taqman 5 nuclease assay, by Thermo Fisher Scientific (1 mentions)
Premade taqman gene expression assays, by Thermo Fisher Scientific (1 mentions)
Mm01231183 m1, by Thermo Fisher Scientific (1 mentions)
5 protocols using mm01244861 m1
1
Quantifying Metabolic Gene Expression
2
Gene Expression Analysis in Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Polymerase chain reaction (PCR) for tissues was performed as described previously [24 (link)] using the primers for URAT1 (Applied Biosystems, Mm01236822_m1), UCP1 (Applied Biosystems, Mm01244861_m1), PGC1α (Applied Biosystems, Mm01208835_m1), Dio2 (Applied Biosystems, Mm00515664_m1), chemokine ligand 2 (Ccl2) (Applied Biosystems, Mm00441242_m1), tissue necrosis factor α (TNFα) (Applied Biosystems, Mm00443258_m1), and GAPDH (Applied Biosystems, Mn03302249_g1).
3
Quantitative Real-time PCR Analysis of Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from frozen NRCMs, cardiac fibroblasts and frozen heart tissue using TRIzol reagent (Invitrogen), and quantitative real-time PCR was performed using a StepOnePlus Real-time PCR System and the StepOne Software program (Applied Biosystems), as previously described.7 (link) The RT‒PCR protocol consisted of one cycle at 95°C for 20 s followed by 40 cycles at 95°C for 1 s and 60°C for 20 s using the primers for URAT1 (Applied Biosystems, Mm01244861_m1 and Rn01479630_g1), TNFα (Applied Biosystems, Mm00443258_m1 and Rn99999017_m1), MCP1 (Applied Biosystems, Mm00441242_m1 and Rn00580555_m1), IL-1β (Applied Biosystems, Mm00434228_m1 and Rn00580432_m1), CD68 (Applied Biosystems, Mm00432403_m1), NLRP3 (Applied Biosystems, Mm00840904_m1) and GAPDH (Applied Biosystems, Mm03302249_g1 and Rn01775763_g1). The transcriptional levels were determined using the ΔΔCt method with normalization to GAPDH.
4
Quantitative Real-Time PCR for UCP1 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from the frozen tissues using TRIzol reagent (Invitrogen) and a quantitative real-time PCR was performed using a StepOnePlus Real-time PCR System and the StepOne Software program (Applied Biosystems), as described previously9 (link),55 (link). The real-time PCR protocol consisted of one cycle at 95 °C for 20 s followed by 40 cycles at 95 °C for 1 s and 60 °C for 20 s using the primers for UCP1 (Mm01244861_m1; Applied Biosystems). The transcriptional levels were determined using the ΔΔCt method with normalization to GAPDH (Mm99999915_g1; Applied Biosystems).
5
Hepatocyte RNA Extraction and qPCR Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA extraction and quantitative real-time PCR were assayed according to the protocol of (Jung et al. 2018c) with modification. Total RNA was extracted from the harvested hepatocytes using TRIzol reagent (Invitrogen). Gene expression was measured by quantitative real-time PCR (qPCR) using the fluorescent TaqMan 5′nuclease assay on an Applied Biosystems 7000 sequence detection system (Foster City, CA, USA). qPCR was performed using cDNA as a template, 2× TaqMan Master Mix, and 20× premade TaqMan gene expression assays (Applied Biosystems). The qPCR conditions were as follows: 95°C for 10 min, followed by 40 cycles of 95°C for 15 s and 60°C for 1 min. PCR primer mixes for mouse Ucp-1 (Applied Biosystems; Mm01244861_m1), Pgc1α (Applied Biosystems; Mm01208832_m1), Prdm16 (Applied Biosystems; Mm00712556_m1), Cpt1 (Applied Biosystems; Mm01231183_m1), Aco (Applied Biosystems; Mm00801417_m1) and Fabp3 (Applied Biosystems; Mm02342495_m1) were used. The mRNA expression of β-actin was quantified as an endogenous control using the following primers: 5′-CGATGCTCCCCGGGCTGTAT-3′ and 5′-TGGGGTACTTCAGGGTCAGG-3′.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!