Hiperfect

Manufactured by Qiagen

Sourced in Germany, United States, United Kingdom, France, Switzerland, Spain, China, Netherlands

The HiPerFect is a high-performance transfection reagent for efficient delivery of siRNA, miRNA, and plasmid DNA into a wide range of cell types. It is designed to provide reliable and reproducible transfection results.

Automatically generated - may contain errors

Lab products found in correlation

556 protocols using hiperfect

Targeting FAK with siRNA in Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

siRNAs were obtained from Dharmacon (Dharmacon, GE Healthcare, Layfayette, CO) for the following FAK target sequences: FAK2, 5′-GGGCAUCAUUCAGAAGAUA-3′; FAK3, 5′-UAGUACAGCUCUUGCAUAU-3′; FAK4 5′-GGACAUUAUUGGCCACUGU-3′. Cells (3 × 10⁵ cells per well) were treated with HiPerFect (Qiagen, Valencia, CA) alone; HiPerFect plus 20 nM negative control siRNA (1027310, Qiagen); or HiPerFect plus FAK siRNA2, 3, or 4 (20 nM) according to manufacturer's protocol, incubated for 24 to 48 hours following transfection, and then used for experiments. FAK inhibition by siRNA was confirmed using immunoblotting.

Waters A.M., Stafman L.L., Garner E.F., Mruthyunjayappa S., Stewart J.E., Mroczek-Musulman E, & Beierle E.A. (2016). Targeting Focal Adhesion Kinase Suppresses the Malignant Phenotype in Rhabdomyosarcoma Cells. Translational Oncology, 9(4), 263-273.

+ Open protocol

+ Expand

FAK Silencing in Cell Culture

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Small interfering RNAs (siRNA) were obtained from Qiagen (Qiagen Inc., Valencia, CA) for the following FAK target sequence: 5′-CCGGTCGAATGATAAGGTGTA-3′. Transfection was carried out as previously described [25 (link), 26 (link)]. Briefly, cells were plated (3 × 10⁵ cells per well) on 6-well culture plates and allowed to attach overnight. Cells were treated with HiPerFect® (Qiagen) alone, HiPerFect® plus 20nM Negative Control siRNA (1027310, Qiagen), or HiPerFect® plus FAK siRNA [Hs_PTK2_10 FlexiTube siRNA (NM_005607, Qiagen)] according to manufacturer’s protocol. Cells were incubated for 24 to 48 hours following transfection and then used for experiments. FAK inhibition by siRNA was confirmed using immunoblotting.

Gillory L.A., Stewart J.E., Megison M.L., Waters A.M, & Beierle E.A. (2015). FAK and p53 Synergistically Decrease Neuroblastoma Cell Survival. The Journal of surgical research, 196(2), 339-349.

+ Open protocol

+ Expand

Silencing E11 in MC3T3 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

E11 siRNA and scrambled siRNA stocks (Qiagen) were diluted to 10nM. MC3T3 cells were plated at 8 × 10³ cells/cm² and maintained in reduced serum medium. Cells were transfected as per manufacturer's instructions with complexes of E11siRNA with HiPerFect (Qiagen), while control cells were transfected with either complexes of scrambled siRNA, with HiPerFect; or HiPerFect alone. After 24 hr incubation at 37°C/5%CO₂, FGF‐2 (10 ng/ml) was added for a further 24 hr to the cells containing the siRNA/HiPerFect complexes or the HiPerFect alone.

Ikpegbu E., Basta L., Clements D.N., Fleming R., Vincent T.L., Buttle D.J., Pitsillides A.A., Staines K.A, & Farquharson C. (2018). FGF‐2 promotes osteocyte differentiation through increased E11/podoplanin expression. Journal of Cellular Physiology, 233(7), 5334-5347.

+ Open protocol

+ Expand

Knockdown of Focal Adhesion Kinase via siRNA

Check if the same lab product or an alternative is used in the 5 most similar protocols

Small interfering RNAs (siRNA) were obtained from Qiagen (Qiagen Inc., Valencia, CA) for the following FAK target sequence: 5′-CCGGTCGAATGATAAGGTGTA-3′. Cells were plated (3 × 10⁵ cells per well) and allowed to attach overnight. Cells were treated with HiPerFect® (Qiagen) alone, HiPerFect® plus 20nM Negative Control siRNA (1027310, Qiagen), or HiPerFect® plus FAK siRNA [Hs_PTK2_10 FlexiTube siRNA (NM_005607, Qiagen)] according to manufacturer’s protocol, incubated for 24 to 48 hours following transfection and then used for experiments. FAK inhibition by siRNA was confirmed using immunoblotting.

Megison M.L., Gillory L.A., Stewart J.E., Nabers H.C., Mrozcek-Musulman E, & Beierle E.A. (2014). Inhibition of Focal Adhesion Kinase (FAK) Leads to Abrogation of the Malignant Phenotype in Aggressive Pediatric Renal Malignancies. Molecular cancer research : MCR, 12(4), 514-526.

+ Open protocol

+ Expand

Plasmid expression and siRNA/miRNA transfection

Check if the same lab product or an alternative is used in the 5 most similar protocols

For plasmid expression, cells were transfected with pMT-p53-dsRedXP (originally given by Dr G Lahav, Harvard University, Cyan Fluorescent protein was replaced by dsRedXP) and pMDM-2-MDM-2-YFP (from Dr G Lahav, Harvard University) using Fugene HD (Roche, UK) at 4∶2 reagent per µg DNA ratio for 24 hours. For siRNA transfection, cells were transfected with siRNA directed for p53 [100 nM] or with non-specific siRNA as a negative control [nM] using HiPerfect (Qiagen) for 48 hours expression. For miR-34a mimic expression, cells were transfected with miR-34a mimics [100 nM] or with a house keeping (GAPDH) transfection control using HiPerfect (Qiagen) and expressed for 72 hours before MTS assay or 48 h before harvesting for western-blot.

Fan Y.N., Meley D., Pizer B, & Sée V. (2014). Mir-34a Mimics Are Potential Therapeutic Agents for p53-Mutated and Chemo-Resistant Brain Tumour Cells. PLoS ONE, 9(9), e108514.

+ Open protocol

+ Expand

Ectopic RPA2 Variant Expression and Knockdown

Check if the same lab product or an alternative is used in the 5 most similar protocols

Endogenous RPA2 was replaced with ectopic variants and silenced as previously described (Anantha et al., 2007 (link)). In brief, the desired U2-OS clone was grown for 24 h in medium lacking doxycycline to allow expression of the ectopic RPA2. Knockdown of endogenous RPA2 was accomplished by siRNA transfection using HiPerFect (QIAGEN), with cells tested 72 h after transfection. Custom-synthesized siRNAs (GE Healthcare) used were RPA2, sense 5′-AACCUAGUUUCACAAUCUGUU-3′ and antisense 5′-CAGAUUGUGAAACUAGGUUUU-3′ (targeting the 3′-untranslated region); and PP4R2, sense 5′-UAUACUGAGAGGUCUAAUAUU-3′ and antisense 5′-UAUUAGACCUCUCAGUAUAUU-3′. For RPE cells, cells were first transfected with the siRNA targeting endogenous RPA2 and, 24 h later, transiently transfected with the RPA2 expression plasmids. The RPA2 variants, synthesized by GenScript, were expressed with a C-terminal Myc tag from the pEF6/Myc expression plasmid (Vassin et al., 2004 (link)). Cells were analyzed 48 h after DNA transfection. PALB2-null EUFA1341 cells were rescued for PALB2 by transfecting with a PALB2 expression plasmid. Commercial reagents for the siRNA against PALB2 (catalog no. sc-93396; Santa Cruz Biotechnology, Inc.) and control siRNA (catalog no. 1027280; AllStars Negative Control siRNA; QIAGEN) were used. DNA plasmids were transfected using Effectene (QIAGEN), whereas siRNA transfection used HiPerFect.

Murphy A.K., Fitzgerald M., Ro T., Kim J.H., Rabinowitsch A.I., Chowdhury D., Schildkraut C.L, & Borowiec J.A. (2014). Phosphorylated RPA recruits PALB2 to stalled DNA replication forks to facilitate fork recovery. The Journal of Cell Biology, 206(4), 493-507.

+ Open protocol

+ Expand

Transfection of miRNA and siRNA

Check if the same lab product or an alternative is used in the 5 most similar protocols

One day before transfections, cells (3.0 x10⁴ cells/ml) were plated in 10-cm Petri dishes. Mature miRNA and siRNAs sequences are shown in Supplementary Table S1. OMMs or siRNAs (Sigma-Aldrich) (100 nM final concentration) were mixed with HiPerfect (Qiagen, Germantown, MD) on a 1:2 volume ratio (siRNA/OMM: HiPerfect) and incubated for 15–20 min in serum and antibiotic-free Opti-MEM medium at room temperature. The cell culture media of the cells was replaced with Opti-MEM, and the transfection mixture was added dropwise. Transfected cells were incubated overnight, and the next day cell pellets were collected for subsequent experiments.

Quiñones-Díaz B.I., Reyes-González J.M., Sánchez-Guzmán V., Conde-Del Moral I., Valiyeva F., Santiago-Sánchez G.S, & Vivas-Mejía P.E. (2020). MicroRNA-18a-5p Suppresses Tumor Growth via Targeting Matrix Metalloproteinase-3 in Cisplatin-Resistant Ovarian Cancer. Frontiers in Oncology, 10, 602670.

+ Open protocol

+ Expand

miR-146 Overexpression and Knockdown

Check if the same lab product or an alternative is used in the 5 most similar protocols

For miR-146 overexpression, cells were transfected with HiPerFect (Qiagen) according to the fast-forward protocol with miRNA Mimic (we used for miR-146 overexpression mimic MSY0002809; and for control, the all-star negative control siRNA SI03650318; Qiagen) at a final concentration of 50 nM. For miR-146 KD, cells were transfected with HiPerFect (Qiagen) according to the fast-forward protocol with the miRNA power family inhibitor at a final concentration of 100 nM (hsa-miR-146 miRCURY LNA microRNA Power family inhibitor; and as control, Negative Control A; Exiqon).

Tordonato C., Marzi M.J., Giangreco G., Freddi S., Bonetti P., Tosoni D., Di Fiore P.P, & Nicassio F. (2021). miR-146 connects stem cell identity with metabolism and pharmacological resistance in breast cancer. The Journal of Cell Biology, 220(5), e202009053.

+ Open protocol

+ Expand

Modulation of Astrocyte Glucose Response

Check if the same lab product or an alternative is used in the 5 most similar protocols

Astrocytes were cultured in either normal glucose (5 mM) or high glucose (25 mM) DMEM depending upon the experiment. Before transfection, 100,000 to 200,000 astrocytes were seeded in medium size petri dishes (Falcon cat. no. 35046) in 1.5ml of DMEM containing FBS and antibiotics and the appropriate concentration of glucose. Cells were transfected using HiPerfect (Qiagen, Cat. No. 301705) and the fast-forward protocol for transfection of adherent cells recommended by the manufacturer. Astrocytes grown in high glucose DMEM were treated with the miRNA205 inhibitor (Qigaen Cat. No. MIN0000878) or HiPerfect alone (mock-transfected), whereas astrocytes grown in normal glucose DMEM were treated with the miR-205 mimic (Qiagen, Cat. No. MSY0000878) or HiPerfect alone (mock-transfected). Briefly, 10 μl of 20 μM miR-205 inhibitor and 12μL of HiPerfect or 10 μl of 20 μM miR-205 mimic and 24μl of HiPerfect were diluted in 400μL of culture medium without serum. The mix was incubated for 10 minutes at room temperature to allow the formation of transfection complexes. The complexes were then added to the cells in a drop-wise fashion giving a final volume of 2mL and a final concentration of 100nM for the mimic and inhibitor. The plate was gently swirled to evenly distribute the transfection complexes and cells were incubated at 37°C for three days.

Rivera-Aponte D.E., Melnik-Martínez K.V., Malpica-Nieves C.J., Tejeda-Bayron F., Méndez-González M.P., Skatchkov S.N, & Eaton M.J. (2020). Kir4.1 potassium channel regulation via miR-205 in astrocytes exposed to hyperglycemic conditions.. Neuroreport, 31(6), 450-455.

+ Open protocol

+ Expand

Transfection of miRNA-125a-5p in Lung Cancer Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Routine transfections were carried out according to established protocols either with Hiperfect (Qiagen, USA) or with Turbofect (Thermofisher, USA). Hiperfect was used for transfection of miRNAs or siRNAs and Turbofect was used to transfect plasmids according to manufacturer’s protocol.
The miR-125a-5p mimics and antagomirs and the corresponding negative control (NC) were purchased from Exiqon (Woburn, MA). The mimics and inhibitors had Locked Nucleic Acid (LNA) technology to reduce off target effects and were fluorescently labeled to assess transfection efficiency.
A549 and H460 cells were seeded into 6-well plates at a density of 2 × 10⁵ cells/well and cultured at 37°C for 24 h. A final concentration of 10nM miR-125a-5p mimic, and 5nM miR-125a-5p inhibitor or NC was transfected into target cells using Hiperfect (Qiagen) and Opti-MEM^® I reduced serum medium (Thermo Fisher Scientific, Inc. USA), according to the manufacturer’s protocol.

Ghoshal-Gupta S., Kutiyanawalla A., Lee B.R., Ojha J., Nurani A., Mondal A.K., Kolhe R., Rojiani A.M, & Rojiani M.V. (2018). TIMP-1 downregulation modulates miR-125a-5p expression and triggers the apoptotic pathway. Oncotarget, 9(10), 8941-8956.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!