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Hiperfect

Manufactured by Thermo Fisher Scientific
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The HiPerFect is a high-performance laboratory equipment designed for conducting various scientific experiments and analyses. It is a versatile and reliable instrument that offers precise and consistent results. The core function of the HiPerFect is to provide researchers and scientists with a powerful tool for their laboratory operations.

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Lab products found in correlation

Hiperfect, by Qiagen (4 mentions) Opti mem, by Thermo Fisher Scientific (3 mentions) Collagenase type 1, by Worthington (2 mentions) Lipofectamine and plus reagent, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Dexamethasone (dex), by Merck Group (2 mentions) Cell culture media, by Thermo Fisher Scientific (2 mentions) Rosiglitazone, by Enzo Life Sciences (2 mentions) Recombinant human insulin, by Enzo Life Sciences (2 mentions) Hydrocortisone cortisol, by Merck Group (2 mentions) Si malat1, by Thermo Fisher Scientific (1 mentions) Control sirna, by Thermo Fisher Scientific (1 mentions) Opti mem media, by Thermo Fisher Scientific (1 mentions) Stat3 sirna, by Thermo Fisher Scientific (1 mentions) C12 ie dap, by InvivoGen (1 mentions) Pgl3 vector, by Promega (1 mentions) Synergy h4 plate reader, by Agilent Technologies (1 mentions) Fugene hd transfection reagent, by Promega (1 mentions) Gblock, by Promega (1 mentions) Steady glo luciferase assay system, by Promega (1 mentions)

Close spelling variants of this product

HiPerFect Transfection Reagent (15 citations) HiPerfect reagent (6 citations)

15 protocols using hiperfect

1

Silencing MALAT1 in VSMCs

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For short-term depletion experiments, the following siRNA sequences were used: siMALAT1 (F: 5′-CUAGAAUCCUAAAGGCAAAdTdT-3′ and R: 5′-UUUGCCUUUAGGAUUCUAGUAGdTdT-3′) (Invitrogen, United States); control siRNA (F:5′-UUCUUCGAA CGUGUCACGUTT-3′ and R: 5′-ACGUGACACGUUCG GAGAATT-3′) (Invitrogen, United States). VSMCs was transfected with siRNA oligonucleotides using HiPerFect (Invitrogen, United States) according to the manufacturer's instructions. Subsequently, phalloidin staining, EDU staining, cell migration assays, PCR and western blotting were performed as described.
Song T.F., Huang L.W., Yuan Y., Wang H.Q., He H.P., Ma W.J., Huo L.H., Zhou H., Wang N, & Zhang T.C. (2017). LncRNA MALAT1 regulates smooth muscle cell phenotype switch via activation of autophagy. Oncotarget, 9(4), 4411-4426.
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2

siRNA-mediated PLA2G4A knockdown in H4 cells

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H4 cells were transfected with siRNA specific for PLA2G4A (Sigma-Aldrich, EHU009531) or not-targeting (nt) siRNA control (Sigma-Aldrich, 45-SIC001). Cells were reverse transfected in 12-well tissue culture plates with 2 μL of 20 μM siRNA plus 202 μL of the master mix containing 200 μL OptiMEM media (Invitrogen, 31985070) and 2 uL HiPerfect (Invitrogen, 301705) following manufacturer’s instructions. Cells were added at 1.5 × 105 per well. After 24 h cells were re-plated in 24-well plates at 2 × 104 per well on coverslips for IF and at 4 × 104 for western blot analysis. After additional 48 h cells were treated with C1P or amyloid-β(1-40).
Sarkar C., Jones J.W., Hegdekar N., Thayer J.A., Kumar A., Faden A.I., Kane M.A, & Lipinski M.M. (2019). PLA2G4A/cPLA2-mediated lysosomal membrane damage leads to inhibition of autophagy and neurodegeneration after brain trauma. Autophagy, 16(3), 466-485.
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3

Targeted Inhibition of ATX and STAT3

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To specifically inhibit ATX and STAT3, siRNAs targeting these two proteins were used. The BON1 cells were cultured in 6-well plates (3.0 × 105 cells/well) for 72 h prior to transfection. Subsequently, ATX siRNA (20 μmol/L; Thermo Fisher Scientific) and STAT3 siRNA (20 μmol/L; Thermo Fisher Scientific) were individually transfected into the BON1 cells. Negative control siRNA (20 μmol/L; Thermo Fisher Scientific) was used as a negative control in parallel. Transfections were performed with HiPerFect (Invitrogen) according to the manufacturer’s protocol. Knockdown effectiveness was assessed by Western blot for total ATX and STAT3 levels.
Yang L., Yu X, & Yang Y. (2018). Autotaxin upregulated by STAT3 activation contributes to invasion in pancreatic neuroendocrine neoplasms. Endocrine Connections, 7(12), 1299-1307.
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4

THP-1 and PBMC Stimulation with C12-iE-DAP

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THP-1 or CD14+ PBMC cells were seeded at a density of 0.5×106/mL overnight in a 24-well plate in RPMI-10 prior to exposure to C12-iE-DAP (InvivoGen). Ligand was transfected using HiPerfect (Invitrogen). The transfection complex was prepared by combining 200 μl OPTI-MEM with 6 μL HiPerfect and 1 μL ligand (from a 1 mg/mL stock), then incubated for 20 min at RT followed by dropwise addition to each well. Unless otherwise specified, cells were stimulated with 1 μg C12-iE-DAP. At specified times post treatment with ligand, cells were analyzed by flow cytometry, quantitative RT-PCR or cell lysates were prepared and analyzed by immunoblot.
Rommereim L.M., Akhade A.S., Dutta B., Hutcheon C., Lounsbury N.W., Rostomily C.C., Savan R., Fraser I.D., Germain R.N, & Subramanian N. (2020). A small sustained increase in NOD1 abundance promotes ligand-independent inflammatory and oncogene transcriptional responses. Science signaling, 13(661), eaba3244.
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5

Regulation of NOD1 by miRNAs

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Luciferase reporter constructs were engineered by cloning the WT NOD1 3’-UTR, a NOD1 3’-UTR with the miR-15b/16 binding site mutated (miR15b/16-mut), or NOD1 3’-UTR with miR-15b/16 and miR-106a binding sites mutated (miR-mut) (gBLOCK, IDT) into the pGL3 vector (Promega). For luciferase assays, HEK-293 cells were seeded at 4×104 cells/well in a 48-well plate overnight. Cells were transfected with 200 ng NOD1 WT or miR-15b/16-mut 3’-UTR luciferase reporter constructs and 10 ng eGFP using FuGENE HD transfection reagent (Promega). Where noted cells were co-transfected with either miRNA mimics, LNA inhibitors or TSB at a final concentration of 100 nM using HiPerfect (Invitrogen) for 36–48 h. Cells were washed with PBS and lysed in cell lysis buffer (9803, Cell Signaling Technology). Luciferase activity was measured with the Steady-Glo Luciferase Assay System (Promega) using a microplate reader (Biotek Synergy H4 Plate reader). Luciferase activity was normalized for transfection efficiency (eGFP) and plotted as a fold change over the respective control.
Rommereim L.M., Akhade A.S., Dutta B., Hutcheon C., Lounsbury N.W., Rostomily C.C., Savan R., Fraser I.D., Germain R.N, & Subramanian N. (2020). A small sustained increase in NOD1 abundance promotes ligand-independent inflammatory and oncogene transcriptional responses. Science signaling, 13(661), eaba3244.
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6

miRNA-29c-3p Modulation in Lung Cancer Cells

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LC cells were plated 24 h before transfection in Ham’s F10 medium and transfected at a density of ~50–60% confluency (HiPerFect; Qiagen, Germantown, MD, USA) following the manufacturer’s instructions. In brief, 10 nM of miRNA-29c-3p mimic or miRNA-29c-3p inhibitor with 2.5uL HiPerFect were diluted in 1mL serum-free OPTIMEM medium(Invitrogen), then incubated for 15 min at room temperature and transferred to the appropriate well with LC cells. The cells were then incubated overnight at 37 °C in 5% CO2–95% air. The efficiency of the transfection was confirmed by qPCR. Negative controls consisted of cells with no treatment, non-targeting control for mimic experiments, and inhibitor control (Qiagen, Germantown, MD, USA) for inhibitor experiments. The miRNA-29c-3p mimic are double-stranded RNA oligonucleotides designed to mimic endogenously mature miRNA-29c activity. The miR-29c-3p inhibitors are single-strand RNA oligonucleotides designed to inhibit miRNA activity.
Lopez N.N., Rangan R., Clark A.F, & Tovar-Vidales T. (2021). Mirna Expression in Glaucomatous and TGFβ2 Treated Lamina Cribrosa Cells. International Journal of Molecular Sciences, 22(12), 6178.
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7

Keap1 Silencing by siRNA

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Chemically synthesized Keap1-specific siRNA (5’GAATGATCACAGCAATGAA3’) was obtained from Shanghai GenePharma Co., Ltd. Cells in the logarithmic growth phase were transfected with Keap1 siRNA or scrambled siRNA using Lipofectamine 3000 and HiPerFect (Invitrogen) Transfection reagent according to the manufacturer’s instructions.
Wu L., Pan C., Wei X., Shi Y., Zheng J., Lin X, & Shi L. (2018). lncRNA KRAL reverses 5-fluorouracil resistance in hepatocellular carcinoma cells by acting as a ceRNA against miR-141. Cell Communication and Signaling : CCS, 16, 47.
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8

U-2 OS Cell RNAi Transfection Protocol

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U-2 OS cells were transfected using a reverse transfection protocol with Hiperfect (Qiagen) transfection reagent according to the manufacturer's instructions. In short: 125 μL of a SMARTpool of four siRNAs (25 nM each) was dispensed in a 6-well plate (657165, Greiner bio-one). Transfection mix (10 μL Hiperfect in total volume of 375 μL Opti-MEM [Invitrogen] per well) was added, followed by 30 minutes incubation at room temperature. After this 70,000 U-2 OS cells/well were added in 2 mL. Cells treated with siRNA against genes listed in Fig. 3B were treated again 24 h after cell seeding. Cells were collected 72 h after the first transfection.
Baas R., van Teeffelen H.A., Tjalsma S.J, & Timmers H.T. (2017). The mixed lineage leukemia 4 (MLL4) methyltransferase complex is involved in transforming growth factor beta (TGF-β)-activated gene transcription. Transcription, 9(2), 67-74.
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9

APC Knockdown in RPE Cells

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A duplex scrambled sequence control and two APC siRNA duplex target sequences (5′-rCrGrA rCrArA rGrArG rCrUrA rGrArA rGrArU rArArU rUrCC A-3′ + 5′-rUrGrG rArArU rUrArU rCrUrU rCrUrA rGrCrU rCrUrU rGrUrC rGrArA-3′ and 5′-rUrGrA rCrArA rUrArA rArGrC rArGrA rGrGrA rArGrG rUrGA T-3′ + 5′-rArUrC rArCrC rUrUrC rCrUrC rUrGrC rUrUrU rArUrU rGrUrC rArUrC-3′) were ordered from Integrated DNA Technologies (Coralville, IA). RPE cells were transfected for two consecutive days for 14–16 h with either a combination of 10 nM of each APC sequence (APC condition) or 20 nM of scrambled sequence (control condition) using HiPerfect (Invitrogen, Carlsbad, CA). Cells were imaged in FluroBrite DMEM (Invitrogen) plus 10% FBS (17 cells in each group across two experiments; also see TIRFM and wide-field fluorescence microscopy) and harvested for Western blotting on the same day, 48 h after the last transfection. Knockdown efficiency was assessed using Western blot analysis using the APC-M2 antibody at 1:5000 (generously gifted to us by Kristi Neufeld, University of Kansas, Lawrence, KS) and Coomassie stain as loading control.
Hookway C., Ding L., Davidson M.W., Rappoport J.Z., Danuser G, & Gelfand V.I. (2015). Microtubule-dependent transport and dynamics of vimentin intermediate filaments. Molecular Biology of the Cell, 26(9), 1675-1686.
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10

Targeted Cdc42 and IQGAP1 Silencing

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A small interfering RNA (siRNA) quadriplex against human Cdc42 or IQGAP1 ON-TARGET plus SMARTpool was purchased from Dharmacon (Dharmacon, Thermo Scientific, Waltham, MA). For monitoring off-target effects of RNA interference, ON-TARGET plus non-targeting pool (Dharmacon) was used. Transient transfection of siRNA was performed using HiPerfect (QIAGEN, Toronto, Canada), according to the manufacturer's protocol. Briefly, 5×105 cells were seeded on a 6 cm2 dish in DMEM containing 10% of FBS one day prior to transfection. On the following day, 256 ng of siRNA and 20 ml of HiPerfect were diluted in 100 ml of Opti-MEM (GIBCO) medium and incubated for 10 min at room temperature. After incubation, the siRNA transfection complexes were added to the cells on a 6 cm2 dish containing 4 ml of DMEM with 10% FBS.
Okura H., Golbourn B.J., Shahzad U., Agnihotri S., Sabha N., Krieger J.R., Figueiredo C.A., Chalil A., Landon-Brace N., Riemenschneider A., Arai H., Smith C.A., Xu S., Kaluz S., Marcus A.I., Van Meir E.G, & Rutka J.T. (2016). A role for activated Cdc42 in glioblastoma multiforme invasion. Oncotarget, 7(35), 56958-56975.
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