Starfect high efficiency transfection reagent

Manufactured by GenStar

Sourced in China

StarFect High-efficiency Transfection Reagent is a laboratory product designed for the delivery of nucleic acids, such as plasmid DNA or RNA, into a variety of cell types. It is formulated to facilitate efficient and gentle transfection, enabling effective gene expression or gene silencing experiments.

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Lab products found in correlation

Cfa f5881, by Merck Group (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Polyhigh transfection, by Sangon (1 mentions) Histrap excel column, by Cytiva (1 mentions) Amicon ultrafiltration device, by Merck Group (1 mentions)

7 protocols using starfect high efficiency transfection reagent

Lentiviral Transduction of HFF Cells

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To generate the stable HFF-C and HFF-UL23-HA lines, PCDH or PCDH-UL23-HA and psPAX2 packaging plasmid, Pmd2-G envelope plasmid were transfected into Lenti × 293T using StarFect High-efficiency Transfection Reagent (GenStar, Beijing, China) according to the manufacturer’s instructions. After 48 h and 72 h transfection, the lentiviral particles were collected, clarified through an 0.45 µM filter, and stored at −80 °C. At this point, the lentiviral particles were added to HFF cells. After 24 h infection, the media was removed, and the HFF cells were cultured in complete media, as described above. The infected HFF cells were selected with puromycin (2 µg/mL).
To overexpress or silence APOL1, CMPK2, and LGALS9, Lenti × 293T were transfected with two packaging plasmids (psPAX2, Pmd2.G) together with a pLenti-LUC-3 × Flag-P2A-C-tRFP or pLenti-APOL1-3 × Flag-P2A-C-tRFP, pLenti-CMPK2-3 × Flag-P2A-C-tRFP, pLenti-LGALS9-3 × Flag-P2A-C-tRFP, shAPOL1, shCMPK2, and shLGALS9 plasmid using StarFect High-efficiency Transfection Reagent (GenStar, Beijing, China) as previously described. The lentiviral particles were collected and then infected with 2 × 10⁵ HFF cells per well. After 48 h, cells were incubated with new media to perform additional experiments.

Wang H., Peng W., Wang J., Zhang C., Zhao W., Ran Y., Yang X., Chen J, & Li H. (2023). Human Cytomegalovirus UL23 Antagonizes the Antiviral Effect of Interferon-γ by Restraining the Expression of Specific IFN-Stimulated Genes. Viruses, 15(4), 1014.

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Transfection and Autophagy Induction Assay

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We used StarFect High-efficiency Transfection Reagent (GenStar) to transfect plasmids into cells according to the manufacturer’s instructions. Cells were incubated in EBSS (Gibco) to induce autophagy. Puromycin (P9620), MG132 (C-2211-5MG), doxycycline (Doxy; D9891), BafA1 (H2714), 3-MA (M9281-100MG), LPS (L4391-1MG), cycloheximide from microbes (CHX, C1988-1 g), and Triton X-100 (T9284-500ML) were purchased from Sigma. Recombinant TNF-α (300-01A) was purchased from PeproTech. IOX1 (HY-12304) was purchased from MCE. LSB (50 mM Hepes, 150 mM NaCl, 1 mM EDTA, 10% glycerol, 1.5 mM MgCl₂, and 1% Triton X-100) was used. Hepes (15630-080) was purchased from Invitrogen. NaCl (7647-14-5) was purchased from Guangzhou Chemical Reagent Factory. EDTA (V900106-500G), glycerol (V900122-6X500ML) and MgCl₂ (V900020-6X500G) were purchased from Vetec. MOG(35–55) (T510219-0005) was purchased from Sangon Biotech. CFA (F5881) was purchased from Sigma. PTX (#180) was purchased from ListLabs.

Liu D., Zhao Z., She Y., Zhang L., Chen X., Ma L, & Cui J. (2022). TRIM14 inhibits OPTN-mediated autophagic degradation of KDM4D to epigenetically regulate inflammation. Proceedings of the National Academy of Sciences of the United States of America, 119(7), e2113454119.

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Isolation and Preparation of Toxoplasma gondii Biomaterials

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Toxoplasma gondii gDNA, RNA, and STAg were isolated. First, a 0.5 μm filter was used to avoid the influence of cell debris. Parasites were collected after filtration through centrifugation at 2,000 g for 10 min, and the lysate was incubated with buffer A (150 mM NaCl, 25 mM EDTA, 10% SDS, and protein kinase) overnight. gDNAs were isolated using phenol-chloroform extraction, and RNAs were isolated using TRIzol reagent (Invitrogen). As for STAg, the lysate was suspended in PBS, and repetitive freeze-thawing at 37°C and −196°C five times was performed to ensure the complete release of the Toxoplasma antigens. Then the mixture was centrifuged at 4°C, 12,000 g for 30 min. The supernatant was transferred to another new microcentrifuge tube and stored at −80°C. For gDNA, RNA, and STAg stimulation in cells, depurated T. gondii gDNA and RNA were transfected into cells through StarFect high-efficiency transfection reagent (GenStar) at the indicated time. For T. gondii infection in vitro, purified tachyzoites were administered at a multiplicity of infection (MOI) of 3.

Hu Z., Wu D., Lu J., Zhang Y., Yu S.M., Xie Y., Li H., Yang J., Lai D.H., Zeng K., Jiang H., Lun Z.R, & Yu X. (2022). Inflammasome Activation Dampens Type I IFN Signaling to Strengthen Anti-Toxoplasma Immunity. mBio, 13(6), e02361-22.

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Lentiviral Transduction of Brpf1 Isoforms

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The full-length sequence of mouse Brpf1 isoform 4 (Brpf1a) was generated and cloned to a lentiviral expression vector pTSB02, and isoform 1 (Brpf1b) was generated by Bridge PCR and then cloned to same lentiviral expression vector.
Designing and cloning of shRNA expression cassette into pLKO lentiviral vector followed the protocol. The shRNA sequences against Brpf1a and Mn1 were designed and cloned into the pLKO lentiviral vector. To generate lentiviral particles, the constructed shRNA expression plasmid was co-transfected with packaging plasmids psPAX2 and pVSVG into human embryonic kidney 293T cells using StarFect High-efficiency Transfection Reagent (Genstar). All used plasmids were confirmed by sequencing.

He Q., Hong M., He J., Chen W., Zhao M, & Zhao W. (2019). Isoform-specific involvement of Brpf1 in expansion of adult hematopoietic stem and progenitor cells. Journal of Molecular Cell Biology, 12(5), 359-371.

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Overexpression and Silencing of DNAJA4 in EPC Cells

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When the DNAJA4 plasmid was overexpressed, GFP green fluorescent protein plasmid was used as the reference for transfection efficiency, and StarFect High-efficiency transfection Reagent (GenStar, Beijing, China) was used as the transfection reagent and was transfected according to the proportion specified in the instructions. The transfection efficiency was more than 90%.
Three siRNAs were designed at the front, middle, and back of the DNAJA4 ORF sequence, named siJA4-1, siJA4-2, and siJA4-3. A NC siRNA was designed as a negative control. A FAM-fluorescently labeled siRNA was selected as a reference for transfection efficiency. Synthesized by Shanghai Shenggong Biological Co., LTD. (Sangon Biotech, Shanghai, China). The sequence of DNAJA4 siRNA interference is shown in Table 1. Two siRNA with good effect were selected for the experiment. EPC cells were transfected using PolyHigh transfection (Sangon Biotech, Shanghai, China) at 25 nM. Cells were infected with CGSIV at an MOI of 0.1 and harvested at the indicated time points for RNA or DNA extraction. The targeted siRNA sequences of EPC DNAJA4 mRNA and those targeted by the control RNAi plasmid are shown in Table 1.

Liu Z., Xie D., He X., Zhou T, & Li W. (2022). DNAJA4 Promotes the Replication of the Chinese Giant Salamander Iridovirus. Genes, 14(1), 58.

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Purification of Recombinant RBD Protein

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Expi293F cells were diluted to 1.5 × 10⁶ cells/mL before transfection. Next, 50 μg of RBD constructs were transfected into 50 mL suspension Expi293F with StarFect high-efficiency transfection reagent (Genstar, Beijing, China). Following 4 days of culturing by shaking, media were collected and centrifuged to harvest the supernatant and cleared of debris by 0.45-μm filtration before loading onto a 1 mL HisTrap excel column (Cytiva). The column was washed for 5 column volumes (CVs) with wash buffer (0.2 mol/L PBS, pH = 8.0), and 5 CVs of wash buffer supplemented with 20 mmol/L imidazole to equilibrate the column. Next, samples were loaded in equilibration buffer (0.2 mol/L PBS, pH = 8.0, with 20 mmol/L imidazole), after that, protein was eluted with elution buffer (0.2 mol/L PBS, pH = 8.0, 500 mmol/L imidazole). Elution fractions containing the protein were collected for SDS-PAGE validation. Elution buffer was then changed with PBST (0.2 mol/L, 0.05% tween-20, pH = 8.0) for desalting and concentration using Amicon ultrafiltration devices (Millipore, 10 kDa). The protein concentrations of purified samples were determined based on BSA standard curve protein quantification method using BCA Protein Content Assay Kit (Beyotime, Shanghai, China). Protein samples were flash-frozen in liquid nitrogen and stored at −80 °C.

Wang K., Pan Y., Wang D., Yuan Y., Li M., Chen Y., Bi L, & Zhang X.E. (2023). Altered hACE2 binding affinity and S1/S2 cleavage efficiency of SARS-CoV-2 spike protein mutants affect viral cell entry. Virologica Sinica, 38(4), 595-605.

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Poly(I·C) Transfection in 293T-HEK Cells

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293T-HEK cells were transfected with each plasmid, and poly(I·C) was used for stimulation 24 h after transfection. Poly(I·C) was transfected by dropwise deposition of a mix containing 1 μg of poly(I·C) and StarFect high-efficiency transfection reagent (Genstar, Beijing, China). Transfected cells were harvested 12 h after transfection.

Zhao J., Sun L., Zhao Y., Feng D., Cheng J, & Zhang G. (2021). Coronavirus Endoribonuclease Ensures Efficient Viral Replication and Prevents Protein Kinase R Activation. Journal of Virology, 95(7), e02103-20.

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