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Ne per cytoplasmic and nuclear extraction reagents

Manufactured by Thermo Fisher Scientific
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The NE-PER cytoplasmic and nuclear extraction reagents are a set of solutions designed to separate cellular components, specifically the cytoplasm and nuclei, for further analysis. The reagents facilitate the extraction and isolation of these cellular fractions, enabling researchers to study their respective contents and functions.

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Lab products found in correlation

Primescript rt reagent kit, by Promega (1 mentions) Syber green pcr master mix, by Takara Bio (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) M mlv reverse transcriptase, by Promega (1 mentions) Mouse anti gapdh, by Cell Signaling Technology (1 mentions) Anti hdac1, by Cell Signaling Technology (1 mentions) Anti relb, by Cell Signaling Technology (1 mentions) Rabbit anti ikkα, by Cell Signaling Technology (1 mentions) Anti p65, by Cell Signaling Technology (1 mentions) Anti iκbα, by Cell Signaling Technology (1 mentions) Nitrocellulose membrane, by Bio-Rad (1 mentions) Rna isolation kit, by Tiangen Biotech (1 mentions) Ripa lysis buffer, by Merck Group (1 mentions)

Close spelling variants of this product

NE-PER (82 citations) Cytoplasmic Extraction Reagents (39 citations) NE-PER reagents (38 citations) NE-PER Nuclear (24 citations) NE-PER Nuclear and Cytoplasmic Extractions Reagents (3 citations) Extraction Reagents (2 citations)

5 protocols using ne per cytoplasmic and nuclear extraction reagents

1

RNA Extraction and miRNA Expression Analysis

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Total RNA was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA, USA). The cytoplasmic and nuclear fractions were extracted using NE-PER Cytoplasmic and Nuclear Extraction Reagents (Thermo Scientific). The miRNA was reversed transcribed using M-MLV Reverse Transcriptase (Promega, Madison, WI, USA) and cDNA was synthesized by PrimeScript RT reagent kit (Promega, Madison, WI, USA). The qRT-PCR was performed using SYBER Green PCR Master mix (Takara Bio, Inc.,Shiga, Japan). All the primer sequences are listed in Supplementary Table 1. Gene expression level was calculated with 2-ΔΔCt method.
Zhang Y., Hu J., Qu X, & Hu K. (2023). Circular RNA RSU1 promotes retinal vascular dysfunction by regulating miR-345-3p/TAZ. Communications Biology, 6, 719.
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2

Western Blot Analysis of Cell Extracts

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Total protein extracts from cell lines were obtained as previously described (15 (link)). Cytoplasmic and nuclear extracts was processed using NE-PER cytoplasmic and nuclear extraction reagents (Thermo Scientific) according to manufacturer’s instructions. Conditioned medium was collected by maintaining the cells in serum free medium. Samples were resolved using SDS-PAGE, transferred to nitrocellulose membrane (Bio-Rad) and probed with sheep anti-CITED2 (1:250; R&D Systems), rabbit anti-IKKα, anti-IκBα, anti-p65, anti-RelB, anti-HDAC1 (1:1000; Cell Signaling Technology), mouse anti-GAPDH (1:10,000; kindly provided by Dr. Shanmugasundaram Ganapathy Kanniappan, Johns Hopkins University School of Medicine, Baltimore, MD) or Actin (1:1000; Sigma-Aldrich) antibodies. Membranes were incubated with horseradish peroxidase-conjugated antibody against sheep (1:2000; R&D Systems), rabbit or mouse (1:2000; GE HealthCare) IgG and binding was revealed by chemiluminescence detection (Millipore).
Jayaraman S., Doucet M., Lau W.M, & Kominsky S.L. (2016). CITED2 Modulates Breast Cancer Metastatic Ability Through Effects on IKKα. Molecular cancer research : MCR, 14(8), 730-739.
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3

Cytoplasmic-Nuclear RNA Expression Analysis

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This study utilized the NE-PER Cytoplasmic and Nuclear Extraction Reagents (Thermo Fisher Scientific) to separate the cytoplasmic fraction from the nuclear counterpart in line with specific protocols. Thereafter, total RNA was extracted by adopting the RNA Isolation Kit (Tiangen Biotech, Beijing, China). Afterward, the cytoplasmic-to-nuclear ratio of specific RNA expression was measured through qRT-PCR, with GAPDH being the cytoplasmic reference and U6 being the nuclear reference.
Mi Y.Y., Sun C.Y., Zhang L.F., Wang J., Shao H.B., Qin F., Xia G.W, & Zhu L.J. (2021). Long Non-coding RNAs LINC01679 as a Competitive Endogenous RNAs Inhibits the Development and Progression of Prostate Cancer via Regulating the miR-3150a-3p/SLC17A9 Axis. Frontiers in Cell and Developmental Biology, 9, 737812.
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4

LPS-Stimulated RAW 264.7 Cell Fractionation

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RAW 264.7 cells were seeded at 1 × 106 cells/mL on six-well plates and were treated with RRE and stimulated with LPS for indicated periods. After incubation, cells were harvested by centrifugation and washed twice with ice-cold PBS. To obtain whole cell lysates, pellets were resuspended in radioimmunoprecipitation assay (RIPA) lysis buffer (Millipore, Bedford, MA, USA) containing protease and phosphatase inhibitors. Cell debris was removed after centrifugation. The cytosolic and nuclear proteins were extracted using NE-PER cytoplasmic and nuclear extraction reagents according to the manufacturer's instructions (Thermo Scientific, Rockford, IL, USA).
Jeong Y.H., Oh Y.C., Cho W.K., Yim N.H, & Ma J.Y. (2016). Anti-Inflammatory Effect of Rhapontici Radix Ethanol Extract via Inhibition of NF-κB and MAPK and Induction of HO-1 in Macrophages. Mediators of Inflammation, 2016, 7216912.
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5

Cellular Compartment Fractionation

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Transfected cells were treated with TPA or TNF-α for 3 h, washed with PBS, and pelleted by centrifugation. Nuclear and cytoplasmic extracts were prepared using NE-PER cytoplasmic and nuclear extraction reagents (Thermo Fisher Scientific). Nuclear and cytoplasmic protein fractions were obtained according to the manufacturer’s instructions (Thermo Fisher Scientific).
Hong O.Y., Jang H.Y., Lee Y.R., Jung S.H., Youn H.J, & Kim J.S. (2022). Inhibition of cell invasion and migration by targeting matrix metalloproteinase-9 expression via sirtuin 6 silencing in human breast cancer cells. Scientific Reports, 12, 12125.
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