Acsl3

Manufactured by Thermo Fisher Scientific

Sourced in United States

ACSL3 is a laboratory equipment product from Thermo Fisher Scientific. It is designed for specific biological and chemical analysis applications. The core function of ACSL3 is to perform precise measurements and facilitate data analysis within its intended use case.

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Lab products found in correlation

3 protocols using acsl3

Quantitative RT-PCR and Immunoblotting Protocol

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The mRNA was measured using real-time RT-PCR with Taqman one-step RT-PCR reagents and results were normalized to co-amplified GAPDH. The primers and probes are listed as following: ACSL3 (Hs00244853_m1), MBOAT2 (Hs01027245_m1), ELOVL5 (Hs01094711_m1), ELOVL7 (Hs00405151_m1), SC5D (Hs00999007_m1), NANS (Hs00219054_m1), TMPRSS2 (Hs01120965_m1), NKX3.1 (Hs00171834_m1) (purchased from Applied Biosystems); PSA: forward, 5′-GATGAAACAGGCTGTGCCG-3′, reverse, 5′-CCTCACAGCTACCCACTGCA-3′, probe, 5′-FAM-CAGGAACAAAAGCGTGATCTTGCTGGG-3′; AR-V7: forward: 5′-CGGAAATGTTATGAAGCAGGGATGA-3′, reverse, 5′-CTGGTCATTTTGAGATGCTTGCAAT-3′, probe, 5′-FAM-GGAGAAAAATTCCGGGT-3′. For immunoblotting, cells were lysed with RIPA buffer with protease inhibitors and anti-Ser240/244 phosphorylated S6 (Cell Signaling), anti-AR (Millipore), anti-AR-V7, anti-ELOVL7, anti-β-actin, anti-GAPDH (Abcam), or anti-β-tublin (Upstate) antibodies were used. Gels shown are representative of at least 3 independent experiments.

Han W., Gao S., Barrett D., Ahmed M., Han D., Macoska J.A., He H.H, & Cai C. (2017). Reactivation of androgen receptor-regulated lipid biosynthesis drives the progression of castration-resistant prostate cancer. Oncogene, 37(6), 710-721.

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Gene Expression Analysis of Fatty Acid Metabolism

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Aliquots of the tissue samples were homogenized by Tissue Lyser (QIAGEN, Hilden, Germany) and the total RNA was extracted using RNeasy Mini kit (QIAGEN) according to the manufacturer’s instructions. Next, 2.0 μg of the isolated total RNA was used to synthesize cDNA with SuperScript VILO Mastermix (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. The synthesized cDNA solutions were diluted 5-fold by Tris-EDTA (TE) buffer (pH 8.0, NIPPON GENE Co., Ltd., Tokyo, Japan). Before the measurements, the cDNA solutions were further diluted 10-fold with MILLI-Q water (Millipore Corporation, Darmstadt, Germany) and were used for TaqMan probe-based semi quantitative-real time PCR. The mRNA levels of Acsl1, 3, and 5, Lcad, Cpt1a, and β-actin were measured in duplicate on a 7300 Real-Time PCR System (Applied Biosystems, Waltham, MA, USA) using TaqMan Gene Expression Master Mix (Applied Biosystems) according to the manufacturer’s instructions. The data analysis was performed by a calibration curve method using SDS software (Applied Biosystems) and the results were normalized to actb expressions.
The following primer and TaqMan probe mixtures were obtained from Applied Biosystems: Acsl1 (Rn00563137_m1), Acsl3 (Rn00589037_m1), Acsl5 (Rn00586013_m1), Lcad (Rn00563121_m1), Cpt1a (Rn00580702_m1), and Beta-actin (Rn00667869_m1).

Goda K., Kobayashi A., Takahashi A., Takahashi T., Saito K., Maekawa K., Saito Y, & Sugai S. (2017). Evaluation of the Potential Risk of Drugs to Induce Hepatotoxicity in Human—Relationships between Hepatic Steatosis Observed in Non-Clinical Toxicity Study and Hepatotoxicity in Humans-. International Journal of Molecular Sciences, 18(4), 810.

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Quantification of mRNA Expression Using RT-PCR

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The mRNA was measured using real-time RT-PCR with Taqman one-step RT-PCR reagents and results were normalized to co-amplified GAPDH. The primers and probes are listed as following: ACSL3 (Hs00244853_m1), MBOAT2 (Hs01027245_m1), ELOVL5 (Hs01094711_m1), ELOVL7 (Hs00405151_m1), SC5D (Hs00999007_m1), NANS (Hs00219054_m1), TMPRSS2 (Hs01120965_m1), NKX3.1 (Hs00171834_m1) (purchased from Applied Biosystems, Foster City, CA, USA); PSA: forward, 5′-GATGAAACAGGCTGTGCCG-3′, reverse, 5′-CCTCACAGCTACCCACTGCA-3′, probe, 5′-FAM-CAGGAACAAAAGCGTGATCTTGCTGGG-3′ AR-V7: forward: 5′-CGGAAATGTTATGAAGCAGGGATGA-3′, reverse, 5′-CTGGTCATTTTGAGATGCTTGCAAT-3′, probe, 5′-FAM-GGAGAAAAATTCCGGGT-3′. For immunoblotting, cells were lysed with RIPA buffer with protease inhibitors and anti-Ser240/244 phosphorylated S6 (Cell Signaling, Danvers, MA, USA), anti-AR (Millipore, Billerica, MA, USA), anti-AR-V7, anti-ELOVL7, anti-β-actin, anti-GAPDH (Abcam, Cambridge, MA, USA), or anti-β-tubulin (Upstate, Billerica, MA, USA) antibodies were used. Gels shown are representative of at least 3 independent experiments.

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