Transintro el transfection reagent

Manufactured by Transgene

Sourced in China

TransIntro EL Transfection Reagent is a lipid-based transfection agent designed to facilitate the introduction of nucleic acids, such as DNA or RNA, into mammalian cells. It is formulated to provide efficient and reliable transfection performance.

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Lab products found in correlation

Opti mem medium, by Thermo Fisher Scientific (3 mentions) Pmirglo, by Promega (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (2 mentions) Ecori, by New England Biolabs (2 mentions) Sirna duplex, by Thermo Fisher Scientific (1 mentions) Opti mem, by Thermo Fisher Scientific (1 mentions) 25 cm2 cell culture flask, by Corning (1 mentions) Alexa fluor 488 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Endofree plasmid kit, by Tiangen Biotech (1 mentions) Viafect, by Promega (1 mentions) Penicillin, by Transgene (1 mentions) Streptomycin, by Transgene (1 mentions) Fugene hd transfection reagent, by Promega (1 mentions) Pcmv3 vector, by Sino Biological (1 mentions) Mir 204 5p mimic, by GenePharma (1 mentions) Lv tug1 rnai, by Genechem (1 mentions) Sirna3, by GenePharma (1 mentions) Sirna2, by GenePharma (1 mentions) Hitransg a, by Genechem (1 mentions)

Spelling variants of this product

EL Transfection Reagent (6 citations) Transfection reagent (2 citations)

62 protocols using transintro el transfection reagent

Lentiviral-mediated L1 Overexpression and Knockdown

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Overexpression and knockdown of L1 were induced by a lentiviral system. Briefly, hL1CAM, shL1CAM, and the control vector were designed and purchased from VectorBuilder (Suzhou, China). The corresponding plasmids were transfected into HEK293T cells by using Transintro EL Transfection Reagent (FT201‐01, TransGen Biotech, Beijing, China) to generate lentiviral particles. Thereafter, the supernatant containing viruses was harvested by centrifugation at 4000 × g for 10 min. The stable overexpression (L1‐OE) or knockdown (shL1) of L1 and vector control (VC) groups in U87, T98, and GBM1 glioma cell lines were established by infecting the virus‐containing pellets at 37 °C for 48 h and subsequently selected with 3 μg·mL⁻¹ puromycin for 7 days.
To overexpress or knockdown miR‐143‐3p, the miR‐143‐3p mimic (miR‐143‐3p) and its negative control (NC), miR‐143‐3p inhibitor (in‐miR‐143‐3p) and its negative control (in‐NC), which were designed and purchased from RiboBio (Guangzhou, China), were transfected into different glioma cell lines by using Transintro EL Transfection Reagent (TransGen Biotech). In addition, to inhibit HK2 expression in glioma cells small interfering RNAs (siRNAs) targeting HK2 (siHK2‐1 and siHK2‐2) and its scrambled siRNA control (siNC) were transfected with Transintro EL Transfection Reagent (TransGen Biotech). All primer sequences are listed in Table S1.

Huang Y., Zhu C., Liu P., Ouyang F., Luo J., Lu C., Tang B, & Yang X. (2023). L1CAM promotes vasculogenic mimicry formation by miR‐143‐3p‐induced expression of hexokinase 2 in glioma. Molecular Oncology, 17(4), 664-685.

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siRNA Knockdown of IRF1 in H1299 and Vero Cells

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The siRNA duplexes for IRF1 were purchased from Sangon Biotech (Shanghai, China). The sequence of the siRNA sense strand: GGAAAUUACCUGAGGACAUdTdT; negative control was provided by Sangon Biotech. Transfection of siRNA was carried out using transIntroTM EL Transfection Reagent (Transgen biotech) as follows. H1299 or Vero cells were plated to a 12-well plate the day before transfection, 5 μl of 20 μM siRNA duplex and 2.5 μl TransIntro EL were diluted with 100 μl Opti-MEM (Gibco) per well and incubated for 20 min. Cells were replenished with 400 μl Opti-MEM containing 5% FBS, and the transfection mixture was added to each well dropwise. Virus infection was performed at 24 h post-transfection.

Liu S.Y., Huang M., Fung T.S., Chen R.A, & Liu D.X. (2023). Characterization of the induction kinetics and antiviral functions of IRF1, ISG15 and ISG20 in cells infected with gammacoronavirus avian infectious bronchitis virus. Virology, 582, 114-127.

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Transfection and Virus Propagation in SSN-1 Cells

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6.0 × 10⁵ B7GG cells were seeded in 25 cm² cell culture flask (Costar, Washington, DC, USA) and grown to 80% confluence (9.0 × 10⁵) at 37 °C in 5% CO₂. Cells were co-transfected with the supporting plasmids pRGNNV-T7-RNA2-CMV-eGFP (25 μg) together with pRGNNV-T7-RNA1-CMV-eGFP (25 μg) or the mutant plasmids (25 μg) (pRGNNV-T7-RNA1-B2-M1-CMV-eGFP or pRGNNV-T7-RNA1-B2-M2-CMV-eGFP) using TransIntro^TM EL transfection reagent (FT201-01, TransGen Biotech, Beijing, China) according to the manufacturer’s instructions. At 12 h post-transfection, the medium was replaced by 30 mL of DMEM with 5% FBS and cells were cultured at 27 °C for 3 days. Then cells were freeze-thawed three times and then centrifuged at 10,000× g for 5 min. The supernatant was subsequently incubated with fresh SSN-1 cells at 27 °C for three blind passages (3 days/passage) and the cytopathic effect (CPE) was observed daily.

Lei Y., Xiong Y., Tao D., Wang T., Chen T., Du X., Cao G., Tu J, & Dai J. (2022). Construction of Attenuated Strains for Red-Spotted Grouper Nervous Necrosis Virus (RGNNV) via Reverse Genetic System. Viruses, 14(8), 1737.

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Efficient Knockdown of Targets Using ASOs and siRNAs

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ASOs and siRNAs were synthesized by Sangon Biotech (Shanghai, China). Before transfection, 2 × 10⁵ cells were seeded into a 24-well plate and incubated at 37 °C and 5% CO₂ for overnight. Next, 200 nM of ASO or siRNA were transfected into the cells using TransIntroTM EL Transfection Reagent (Transgene, FT201-01) and cells were collected to extract RNA. For primary cells, ASO or siRNA were transfected with NEON system.

Zhou T., Zhu X., Ye Z., Wang Y.F., Yao C., Xu N., Zhou M., Ma J., Qin Y., Shen Y., Tang Y., Yin Z., Xu H., Zhang Y., Zang X., Ding H., Yang W., Guo Y., Harley J.B., Namjou B., Kaufman K.M., Kottyan L.C., Weirauch M.T., Hou G, & Shen N. (2022). Lupus enhancer risk variant causes dysregulation of IRF8 through cooperative lncRNA and DNA methylation machinery. Nature Communications, 13, 1855.

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DTMUV Infection Protein Analysis in Transfected Cells

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Transfection of plasmid DNA into cells was performed with the TransIntroTM EL Transfection Reagent (Transgen, Beijing, China), as previously described [26 (link)]. At 24 h post-transfection, cells were infected with DTMUV at an MOI of 1 or mock-treated with UV-DTMUV and cultured in FBS-free media until harvest for the extraction of proteins and/or RNA at the indicated time post-infection.
Proteins in the total cell lysates were separated by SDS-PAGE and transferred to a nitrocellulose membrane. The membrane was probed by incubation with primary antibodies and a secondary antibody conjugated with fluorescein isothiocyanate, and protein bands were detected with an Azure c600 imager. Densitometric quantification of band density was performed using the NIH programme Image J (https://imagej.nih.gov/ij/) (accessed on 18 October 2021). Each experiment was performed three times with comparable findings, and a single typical result is displayed.

Xiang C., Yang Z., Xiong T., Wang T., Yang J., Huang M., Liu D, & Chen R. (2022). Avian IRF1 and IRF7 Play Overlapping and Distinct Roles in Regulating IFN-Dependent and -Independent Antiviral Responses to Duck Tembusu Virus Infection. Viruses, 14(7), 1506.

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Plasmid Transfection and Capsid Protein Expression

Cited 1 time

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Plasmids pVAX-C and pVAX were prepared using the EndoFree Plasmid Kit (Tiangen, Beijing, China) and were transfected into COS7 cells using the TransIntro^TM EL Transfection Reagent (TransGen Biotech, Beijing, China) when the cells were growing at around 80% confluent in six-well plates. The expression of the capsid protein in the cells post 48 h transfection was confirmed by indirect immunofluorescence as described previously [21 (link)]. Briefly, transfected cells were washed with phosphate-buffered-saline (PH 7.2) (PBS), then fixed with 4% paraformaldehyde and permeabilized with 0.2% Triton X-100 in PBS and blocked with 5% BSA in PBS (BSA-PBS). After that, the cells were incubated with rabbit anti-capsid protein polyclonal antibody (prepared by our lab) as the primary antibody diluted 1:100 in BSA-PBS for 2 h, followed by incubation with the Alexa Fluor 488-conjugated goat anti-rabbit IgG (Thermo Fisher, Lafayette, CO, USA) as the secondary antibody was diluted 1:2000 in BSA-PBS for 2 h at room temperature. The cell nuclei were counterstained with 4′, 6-diamidino-2-phenylindole (DAPI) for 10 min at room temperature. The cells were examined by fluorescence microscopy (Nikon, Tokyo, Japan).

Huang J., Shen H., Jia R., Wang M., Chen S., Zhu D., Liu M., Zhao X., Yang Q., Wu Y., Liu Y., Zhang L., Yin Z., Jing B, & Cheng A. (2018). Oral Vaccination with a DNA Vaccine Encoding Capsid Protein of Duck Tembusu Virus Induces Protection Immunity. Viruses, 10(4), 180.

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STAT3 Knockdown in SK-HEP-1 Cells

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SK-HEP-1 cells were seeded onto 6-well plates and transfected with siSTAT3 (siSTAT3 sense: 5-GAAUCAAGCAGUUUCUUCATT-3; siSTAT3 antisense: 5-UGAAGAAACUGCUUGAUUCTT-3, Tsingke Biotechnology, Beijing, China) by TransIntro^TM EL Transfection Reagent (TransGen Biotech) for 48 h. Thereafter, the cells were harvested for western blot and RT-qPCR.

Peng T., Wonganan O., Zhang Z., Yu J., Xi R., Cao Y., Suksamrarn A., Zhang G, & Wang F. (2021). A 2-Benzylmalonate Derivative as STAT3 Inhibitor Suppresses Tumor Growth in Hepatocellular Carcinoma by Upregulating β-TrCP E3 Ubiquitin Ligase. International Journal of Molecular Sciences, 22(7), 3354.

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Cell Culture and Transfection Protocols

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C2C12 myoblasts and NIH3T3 cells were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA) and CAS (Chinese Academy of Sciences, Beijing, China), respectively. HEK293T cells were kindly provided by Prof. Jianhua Wu (South China University of Technology, Guangzhou, China). The C2C12 myoblasts, NIH3T3 and HEK293T cells were all maintained in high-glucose DMEM, supplemented with 10% FBS (HyClone, Logan, UT, USA), 2 mM L-Glutamine, 100 U/mL penicillin and 100 μg/mL streptomycin (Transgen Biotech, Beijing, China) (growth medium, GM) in a 5% CO₂ incubator at 37 °C. The C2C12 myoblasts (passage 7~10) were transfected using Via-FectTM (Promega, Madison, WI, USA), while the NIH3T3 and HEK293T cells were transfected with TransIntroTM EL Transfection Reagent (Transgen Biotech, Beijing, China). For C2C12 myoblast differentiation, the cell-cultured medium was switched to DMEM containing 2% horse serum, 100 U/mL penicillin and 100 μg/mL streptomycin (differentiation medium, DM) once the C2C12 myoblasts became confluent.

Xiong Z., Wang M., You S., Chen X., Lin J., Wu J, & Shi X. (2022). Transcription Regulation of Tceal7 by the Triple Complex of Mef2c, Creb1 and Myod. Biology, 11(3), 446.

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Dual-luciferase Reporter Assay for miRNA

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The dual-luciferase reporter assay was performed as described previously (15 (link)). In brief, SSN-1 cells were co-transfected with NC mimic, miR-214 mimic, NC inhibitor, or miR-214 inhibitor, together with the luciferase reporter plasmids using TransIntroTM EL Transfection Reagent (TransGen Biotech, China). At 24 h post of transfection, the Renilla and firefly luciferase activities were measured, and the data were expressed as relative firefly luciferase activity normalized to Renilla luciferase activity.

Zhang C., Feng S., Zhang W., Chen N., Hegazy A.M., Chen W., Liu X., Zhao L., Li J., Lin L, & Tu J. (2017). MicroRNA miR-214 Inhibits Snakehead Vesiculovirus Replication by Promoting IFN-α Expression via Targeting Host Adenosine 5′-Monophosphate-Activated Protein Kinase. Frontiers in Immunology, 8, 1775.

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Cloning and Expressing Autophagy Genes

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Full-length of BmAtg3, BmAtg4, BmAtg7, and BmAtg8 were cloned from total cDNA obtained from prepupal B. mori fat body and fused with c-Myc, His, V5, HA, or FLAG, respectively. Then they were inserted into the multiple cloning sites of pIEX4 vectors (Novagen, Beijing, China, 71235-2). The HsATG4a, b, c, d fused with tag sequence were cloned and constructed into pcMV3 vectors (Sino Biological, Beijing, China, HG22147-UT, HG20407-CY, HG15081-CM, and HG15537-CY). BmN or HEK293 cells were transiently transfected with the respective plasmids using FuGENE® HD Transfection Reagent (Promega, WI, USA, E2311) or TransIntro^TM EL Transfection Reagent (TransGen Biotech, Beijing, China, FT201-01), according to the manufacturer’s instruction.

Wu W., Li K., Guo S., Xu J., Ma Q., Li S., Xu X., Huang Z., Zhong Y., Tettamanti G., Cao Y., Li S, & Tian L. (2021). P300/HDAC1 regulates the acetylation/deacetylation and autophagic activities of LC3/Atg8–PE ubiquitin-like system. Cell Death Discovery, 7, 128.

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