Hiperfect transfection reagent

Manufactured by Qiagen

Sourced in Germany, United States, China, Italy, United Kingdom, France, Spain, Netherlands, Japan, Canada

HiPerFect Transfection Reagent is a lipid-based transfection reagent designed for efficient delivery of nucleic acids into mammalian cells. It facilitates the uptake of DNA, RNA, and other molecular constructs into the target cells.

Automatically generated - may contain errors

Lab products found in correlation

1 417 protocols using hiperfect transfection reagent

Transfection Assay for miRNA Targeting

Cited 1 time

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

TargetScan [9 ] was used to predict biological targets of micro RNAs (miRNAs) by searching for the presence of conserved sequenced corresponding to miRNA binding site. P_CT (probability of conserved targeting) estimates the probability of the site being preferentially conserved because it is targeted by the cognate miRNA [10 (link)].
For the transfection assay, 6 × 10⁵ skin fibroblasts were seeded in each well of 4-well plates (21.8 cm²/well). Transfection was performed using Hiperfect Transfection Reagent (Qiagen S.A., Courtaboeuf, France), 150 ng miScript miRNA Mimic (Syn-hsa-miR-9-5) (Qiagen S.A., Courtaboeuf, France) or 1500 ng miScript miRNA Inhibitor (Anti-hsa-miR-9-5) (Qiagen S.A., Courtaboeuf, France), according to the manufacturer’s recommendation. For controls, transfection control (Hiperfect Transfection Reagent only) and a negative control (1500 ng miScript Inhibitor Negative Control; Qiagen S.A., Courtaboeuf, France) had been tested. Each condition had been tested in three biological replicates.
Total RNA were extracted 72 h after transfection with the miRNeasy kit^® according to the manufacturer’s instructions. After purification, RNAs were eluted in 15 µL of RNase-free water and the concentrations of RNA were estimated by measuring absorbance at 260 nm, using Nanodrop 2000/2000 c system. Reverse transcription was performed as above.

Benarroch L., Aubart M., Gross M.S., Jacob M.P., Arnaud P., Hanna N., Jondeau G, & Boileau C. (2018). Marfan Syndrome Variability: Investigation of the Roles of Sarcolipin and Calcium as Potential Transregulator of FBN1 Expression. Genes, 9(9), 421.

+ Open protocol

+ Expand

Evaluation of GPNMB and STAT1 siRNA Knockdown

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

Three GPNMB siRNA sequences were examined for their effects on the downregulation of GPNMB expression, and all the three sequences were shown to have ability to interfere GPNMB mRNA (Supplementary Fig. S2). However, the siRNA no. 21 was used for the subsequent analysis because it was found to be the most effective. NHEKs and NHEMs were transfected with 5 nM GPNMB siRNA (No. 21) and a negative control siRNA (FlexiTube siRNA; Qiagen, CA, USA) using HiPerFect transfection reagent (Qiagen, CA, USA) according to the manufacturer’s instructions. The knockdown was verified by western blotting with an antibody specific for GPNMB. The siRNA sequences (No. 21) of GPNMB were as follows: 5′-GGAGCUGAGUAGGAUUCCUGAUGAA-3′ (forward) and 5′-UUCAUCAGGAAUCCUACUCAGCUCC-3′ (reverse). On the other hand, the STAT1 siRNA at 5 nM level and a negative control siRNA (FlexiTube siRNA; Qiagen, CA, USA) were transfected into NHEKs using HiPerFect transfection reagent (Qiagen, CA, USA) according to the manufacturer’s instructions.

Biswas K.B., Takahashi A., Mizutani Y., Takayama S., Ishitsuka A., Yang L., Yang F., Iddamalgoda A., Katayama I, & Inoue S. (2020). GPNMB is expressed in human epidermal keratinocytes but disappears in the vitiligo lesional skin. Scientific Reports, 10, 4930.

+ Open protocol

+ Expand

MicroRNA Transfection in Liver Cells

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

The miR mimics, inhibitors and negative control (NC) were purchased from Genepharma (Shanghai, China), as were the siRNA-JUN and the non-specific siRNA (NC). The miR and siRNA transfections were performed using the HiPerfect transfection reagent (QIAGEN) as previously described [44 (link)]. Briefly, 1 × 10⁶ cells in 2 ml of L-DMEM culture medium supplemented with serum and antibiotics were seeded in 6-well plates. At the same time, the miR mimics, inhibitors or NC were mixed with HiPerfect transfection reagent (QIAGEN, Duesseldorf, Germany) and incubated at room temperature for 10 min. The NCTC1469 and Hep1-6 cells were incubated with the complex for 48 h.

Guo J., Fang W., Sun L., Lu Y., Dou L., Huang X., Sun M., Pang C., Qu J., Liu G, & Li J. (2016). Reduced miR-200b and miR-200c expression contributes to abnormal hepatic lipid accumulation by stimulating JUN expression and activating the transcription of srebp1. Oncotarget, 7(24), 36207-36219.

+ Open protocol

+ Expand

Transfection of MSCs with miRNA Mimics

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

MSCs were transiently transfected with miR-200a, miR-200b, miR-200c, miR-141 and miR-429 mimics (20 μM, MiScript miRNA mimics, Qiagen) using the HiPerFect Transfection Reagent (Qiagen), according to the manufacturer’s protocol. Briefly, MSCs were seeded into 6-well plates at a density of 10,000 cells/cm². MiRNA mimics at a concentration of 20 nM were mixed with an appropriated volume of HiPerFect Transfection Reagent (Qiagen) and incubated at room temperature for 10 minutes. Allstars negative control siRNA (20 μM, Qiagen) was used as scrambled control. The transfection mix was added dropwise to the cells in FBS-depleted culture medium, in agreement with the manufacturer's instructions. After 24-hours of incubation, the transfection medium was removed, and cells were cultured in MSCBM for 7 days.

Chiabotto G., Bruno S., Collino F, & Camussi G. (2016). Mesenchymal Stromal Cells Epithelial Transition Induced by Renal Tubular Cells-Derived Extracellular Vesicles. PLoS ONE, 11(7), e0159163.

+ Open protocol

+ Expand

siRNA Knockdown of Transcription Factors

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

siRNA knockdown was performed by wet-reverse transfection using HiPerFect transfection reagent according to manufacturer’s protocol (Qiagen, HiPerFect transfection reagent, cat no 301704). In brief, siRNA (50 nM from Ambion: siRNA-Sp1 ID: 116546, siRNA-AP1/JUN ID: 145018, siRNA-AP-1/Fos ID: 115631; and Sigma: siRBA-E2F1 cat no. EHU070981) and HiPerFect Reagent were mixed and spotted into wells. After complex formation (15 min incubation), cells were plated on the mixture in their respective media and incubated for 72 h before assessing the efficiency of knock down. See Supplementary Fig. 3b for knockdown efficiency validation using immunoblot analysis of E2F1, Sp1 and AP-1 expression.

Rasmussen R.D., Gajjar M.K., Tuckova L., Jensen K.E., Maya-Mendoza A., Holst C.B., Møllgaard K., Rasmussen J.S., Brennum J., Bartek J J.r., Syrucek M., Sedlakova E., Andersen K.K., Frederiksen M.H., Bartek J, & Hamerlik P. (2016). BRCA1-regulated RRM2 expression protects glioblastoma cells from endogenous replication stress and promotes tumorigenicity. Nature Communications, 7, 13398.

+ Open protocol

+ Expand

Silencing Experiments in CaCo-2 Cells

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

Silencing experiments were performed with two different silencing mRNAs for HRS (Hs HRS 5 and Hs HRS 6), MyD88 (Hs MYD88 2 and Hs MYD88 8), TLR7 (Hs TLR7 6 and Hs TLR7 8) with similar results. Non-specific siRNA (MAPK1) was used for transfection efficiency (not shown) and scrambled mRNA sequences was used to test specificity of the silencing (All Stars Negative). All silencing mRNAs were all purchased from QIAGEN, Milan, Italy. Transfections were carried out using the HIPerFect Transfection Reagent following the manufacturer’s instructions (QIAGEN, Milan, Italy).
Briefly, CaCo-2 cells were incubated in standard growth conditions, and 6 μg of siRNA was diluted in 1 ml of culture medium without serum to give a final siRNA concentration of 50 nM. Twenty microliters of HIPerFect Transfection Reagent were added to the siRNA mixture with vortexing, and the transfection mixture was added drop wise onto the cells and incubated for 72 h. The cells were then processed for immunoprecipitation and for Western blot (WB) analysis.

Nanayakkara M., Lania G., Maglio M., Auricchio R., De Musis C., Discepolo V., Miele E., Jabri B., Troncone R., Auricchio S, & Barone M.V. (2018). P31–43, an undigested gliadin peptide, mimics and enhances the innate immune response to viruses and interferes with endocytic trafficking: a role in celiac disease. Scientific Reports, 8, 10821.

+ Open protocol

+ Expand

MiRNA Transfection in hDFC Cells

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

For transfection of miRNA, HiPerFect Transfection Reagent (QIAGEN) was used in this study, in accordance with the manufacturer's protocol. Briefly, hDFC were seeded at 3.1 × 10³ cells/cm² in GM. After 24 h (day 0), media were replaced with OIM, then the transfection mixture was added. The transfection mixture contained 0.6% HiPerFect Transfection Reagent, and 5 nM miR-204-5p mimic (QIAGEN) or 5 nM AllStars Negative Control siRNA (QIAGEN) in MSC basal medium. Transfection was performed on day 0 and day 3.

Ito K., Tomoki R., Ogura N., Takahashi K., Eda T., Yamazaki F., Kato Y., Goss A, & Kondoh T. (2020). MicroRNA-204 regulates osteogenic induction in dental follicle cells. Journal of Dental Sciences, 15(4), 457-465.

+ Open protocol

+ Expand

Transfection of miRNA Modulators in Immune Cells

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

The transfection of miRNA mimics, inhibitors, and controls (GenePharma) were performed with HiperFect Transfection Reagent (Qiagen). Briefly, 20 µM mimics or inhibitors were transfected into THP-1, THP-1-MA cells for 48 h. Subsequently, the THP-1 cells were stimulated using lipopolysaccharide (LPS; 1 µg/mL; Sigma), and incubated for a further 24 h. The cells were collected for RNA extraction, and the supernatants were collected for IP-10 detection. Transfection of primary CD14⁺ monocytes with mimics was performed with RNAiMAX (Invitrogen) according to the manufacturer’s protocol. The sequences of the mimics and inhibitors are listed in Table S1 in Supplementary Material. The forced reduction process of ISG15 was achieved by employing the 20 µM ISG15 siRNA for 24 h (Invitrogen). This was followed by THP-1 cell treatment with PMA for 48 h, into which was transfected 20 nM mimics by HiperFect Transfection Reagent (Qiagen). For the inhibition of ISG15 in THP-1 cells, IG15 siRNA was transfected to THP-1 cells for 24 h. The siRNA control was represented by non-specific Stealth RNAi^® Negative Control Duplexes.

Wu X., Zhang L.L., Yin L.B., Fu Y.J., Jiang Y.J., Ding H.B., Chu Z.X., Shang H, & Zhang Z.N. (2017). Deregulated MicroRNA-21 Expression in Monocytes from HIV-Infected Patients Contributes to Elevated IP-10 Secretion in HIV Infection. Frontiers in Immunology, 8, 1122.

+ Open protocol

+ Expand

miR-542-5p Overexpression Impacts Cell Viability

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

1×10⁵ cells/well were seeded into 24-well microplate with DMEM. 18 h after seeding, 1 μL HiPerFect transfection reagent (Qiagen, Shanghai, China) with 2 μL 20 nM negative control or hsa-miR-542-5p mimics (GenePharma, Shanghai, China) were used for transfecting, mixed with 60 μL DMEM for 10 min, then medium was removed from the culture container, and the mixture above added to 150 μL medium with cells. Cells were harvested at 48 h after transfection. The transfection efficiency of hsa-miR-542-5p was confirmed by RT-PCR.
For cell viability detection, 1×10⁴ cells/well were seeded into a 96-well microplate, 18 h after seeding, 0.3μL HiPerFect transfection reagent (Qiagen, Shanghai, China) with 0.6 μL 20 nM negative control or hsa-miR-542-5p mimics (GenePharma, Shanghai, China) were used for transfecting, mixed with 20 μL DMEM for 10 minutes, then removed medium from culture container, and the mixture above added to 100 μL medium with cells. After 24 h, cell activity was measured as described above, negative control was set as control.

Cheng S., Zhang Z., Hu C., Xing N., Xia Y, & Pang B. (2020). Pristimerin Suppressed Breast Cancer Progression via miR-542-5p/DUB3 Axis. OncoTargets and therapy, 13, 6651-6660.

+ Open protocol

+ Expand

Evaluating miRNA Transfection Efficacy in SLK Cells

Cited 1 time

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

HiPerFect Transfection Reagent (Qiagen, Hilden, Germany) and AllStars Hs Cell Death Control siRNA (Qiagen) were mixed according to the manufacturer's instructions. The mixture was then added to 96-well plates containing SLK cell cultures and an MTT Cell Proliferation Cytotoxicity Assay Kit (Sang Biotech, Shanghai, China) was used to measure the cell viability at 6, 24, 48, and 72 hours, respectively, by measuring the absorbance at 450 nm using a spectrophotometer (NanoDrop, NY). Six wells were measured for each time point and all experiment were done in triplicate. Cell viability (%) = (As – Ab)/(Ac – Ab) × 100%, with data expressed as the mean (Ab: blank control group; Ac: negative control group; As: experimental group).^{[6 (link)]}According to the above optimal transfection conditions, the miR-126 mimic (miR-126m, cat. no. MSY0000445), miR-126 inhibitor (miR-126i, cat. no. MIN0000445), AllStars Negative Control siRNA (miR-126NC, cat. no.1027272), and miScript Inhibitor Negative Control miR-126 (miR-126iNC, cat no.1027271) were purchased from Qiagen (Qiagen). Cells were transfected with HiPerFect Transfection Reagent (Qiagen) following the manufacturer's instructions. SLK cells were used as blank control to exclude the effect of small nucleotide fragments on SLK cells.

Lu G., Wu X., Zhao Z., Ding Y., Wang P., Wu C., Kang X, & Pu X. (2018). MicroRNA-126 regulates the phosphatidylinositol-3 kinase (PI3K)/protein kinase B (AKT) pathway in SLK cells in vitro and the expression of its pathway members in Kaposi's sarcoma tissue. Medicine, 97(35), e11855.

+ 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!