Frozen heart tissue was homogenized with an IKA T25D ULTRA TURRAY homogenisator (Laboratory equipment, Germany) in Trizol, followed by chloroform extraction and isopropanol precipitation. Next, RNA was DNase treated with the NucleoSpin RNA II Kit (Macherey‐Nagel, Düren, Germany) and subsequently reverse transcribed via the High Capacity cDNA Reverse Transcription Kit from Applied Biosystems by Thermo Fisher Scientific (Carlsbad, CA, USA). To assess the mRNA expression of the target genes MCP‐1, MCP‐3, CCL5, stromal cell‐derived factor‐1α (SDF‐1α), IL‐6, IL‐12, TNF‐α, transforming growth factor‐ß (TGF‐ß), VCAM‐1, and intercellular adhesion molecule‐1 (ICAM‐1) mRNA expression was analyzed via real‐time PCR using gene expression assays for MCP‐1 Mm00438270_m1, MCP‐3 Mm00443113_m1, CCL5 Mm01302428_m1, SDF‐1α Mm00445552_m1, IL‐6 Mm00446190_m1, IL‐12 Mm00434165_m1, TNF‐α Mm00443258_m1, TGF‐ß1 Mm00441724_m1, VCAM‐1 Mm01320970_m1, and ICAM‐1 Mm00516023_m1 from Applied Biosystems by Thermo Fisher Scientific (Carlsbad, CA), respectively. mRNA expression was normalized to the housekeeping gene CDKN1b Mm00438167_g1 and relatively expressed with the control group set as 1.
Vcam 1 mm01320970 m1
Manufactured by Thermo Fisher Scientific
VCAM-1 Mm01320970_m1 is a TaqMan Gene Expression Assay designed to detect and quantify the expression of the mouse vascular cell adhesion molecule 1 (VCAM-1) gene. This assay can be used for gene expression analysis in mouse samples using real-time PCR or reverse transcription PCR (RT-PCR) techniques.
Automatically generated - may contain errors
Lab products found in correlation
Mm00446190 m1, by Thermo Fisher Scientific (1 mentions)
Mm00445552 m1, by Thermo Fisher Scientific (1 mentions)
Tnfα mm00443258 m1, by Thermo Fisher Scientific (1 mentions)
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
Nucleospin rna 2 kit, by Macherey-Nagel (1 mentions)
Ddit3, by Thermo Fisher Scientific (1 mentions)
Icam 1, by Thermo Fisher Scientific (1 mentions)
Ch25h, by Thermo Fisher Scientific (1 mentions)
Ht7900 abi sequence detection system, by Thermo Fisher Scientific (1 mentions)
Taqman pcr mix, by Thermo Fisher Scientific (1 mentions)
Taqman reverse transcription reagent, by Thermo Fisher Scientific (1 mentions)
Rneasy kit, by Qiagen (1 mentions)
2 protocols using vcam 1 mm01320970 m1
1
Analyzing Inflammatory Gene Expression
2
Quantitative Gene Expression Analysis
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RNA was isolated by RNeasy Kit (Qiagen). Complementary DNAs were generated from 1.5 μg of purified RNA using TaqMan reverse transcription reagents from Applied Biosystems. Real-time PCR was performed in triplicate with TaqMan PCR Mix (Applied Biosystems) in the HT7900 ABI sequence Detection System (Applied Biosystems). Predesigned primers were purchased from Applied Biosystems (Vcam1 [Mm01320970_m1], Atf4 [Mm00515325_g1], Ddit3 [Mm01135937_g1], Ch25h [Mm00515486_s1], and Icam1 [Mm00516023_m1]), using Gusb (Mm00446953_s1) or Gapdh (Mm99999915_g1) as a housekeeping gene for normalization.
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