Neurofect transfection reagent

Manufactured by Genlantis

Sourced in United States

Neurofect is a transfection reagent designed for efficient delivery of nucleic acids, such as plasmid DNA, siRNA, and mRNA, into a variety of neuronal and glial cell types. It facilitates the uptake of these molecules into the target cells, enabling gene expression or gene silencing studies.

Automatically generated - may contain errors

Lab products found in correlation

Vernier stereotaxic instrument, by Leica (3 mentions) Shc002, by Merck Group (1 mentions) Plko 1 trc cloning vector, by Addgene (1 mentions) Siglo green, by Genlantis (1 mentions) Stereotaxic device, by Stoelting (1 mentions) B 002000 ub 100, by Horizon Discovery (1 mentions)

5 protocols using neurofect transfection reagent

Stereotaxic Delivery of siRNA in Mice

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

Stereotaxic injection of siRNA was performed as described previously (Choi et al., 2015). In brief, diluted siRNA (50 ng/μl) was mixed with Neurofect transfection reagent (T800075; Genlantis, San Diego, CA, USA). The siRNA mix (1.8 μl of 7.5 ng/μl) was injected into each CA3 (stereotaxic coordinate: AP, −1.9; ML, ±3.0; DV, −2.1 mm) using a stereotaxic injection system (Vernier Stereotaxic Instrument, Leica Biosystems, Wetzlar, Germany).

Choi J., Kwon H., Lee J., Lee Y., Seoh J, & Han P. (2019). Hyperoxygenation revitalizes Alzheimer’s disease pathology through the upregulation of neurotrophic factors. Aging Cell, 18(2), e12888.

+ Open protocol

+ Expand

Knockdown of Sp4 in Primary Neurons

Cited 2 times

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

To knockdown Sp4 expression, a vector-based shRNA approach was used, and two target sequences were chosen from the RNAi Consortium’s Public TRC Cloning Database at the Broad Institute (Table 4) and each cloned into the pLKO.1 TRC cloning vector (Plasmid 10878; Addgene, Cambridge, MA, USA) as reported previously [5 (link)]. To rule out the non-specific consequences of shRNA, the pLKO.1 non-mammalian shRNA control vector SHC002 (Sigma), a scrambled negative control, was also used.
Transfection of primary neurons was carried out 4 days post-plating with both Sp4 shRNA constructs (2 μg of each construct) or the pLKO.1 non-mammalian control (2 μg), using Neurofect transfection reagent per 6-well plate according to the manufacturer's instructions (Genlantis). TurboGFP (0.5 μg) vector was added to each well for transfection visualization and selection efficiency. Transfection efficiency was around 50–60% before selection. Puromycin selection, however, effectively yielded 100% transfected cells. Transfection efficiency was observed using green fluorescence. Primary neurons transfected with shRNA against Sp4 were further stimulated with KCl. A final concentration of 20 mM KCl was added to the culture medium for 5 h according to our published method [2 (link)]. After this period, cells were harvested for RNA and protein isolation.

Nair B., Johar K., Priya A, & Wong-Riley M.T. (2015). Specificity protein 4 (Sp4) transcriptionally regulates inhibitory GABAergic receptors in neurons. Biochimica et biophysica acta, 1863(1), 1-9.

+ Open protocol

+ Expand

Stereotaxic siRNA Delivery for Brain Research

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

Stereotaxic injection of siRNA was carried out as described previously [20 (link)30 (link)31 (link)]. Mice were anesthetized with a mixture of ketamine hydrochloride (50 mg/mL) and xylazine hydrochloride (23.3 mg/ml) at a dose of 2.5 µl per body weight (g). While the mouse head was held in the stereotaxic device (Stoelting Company, Wood Dale, IL, USA), 1.5 µl (18 ng of siRNA) of siRNA mixture per each side of the dorsal striatum (AP, +1.0; ML, ±1.5; DV, −3.6 mm) was injected at the rate of 0.2 µl/min using a 30 G needle. The siRNA mixture was prepared 20 min before injection by mixing 1 µl of control-siRNA or TrkB-siRNA with siGLO Green (1/19 of target gene-siRNA), 0.5 µl of 50% sucrose and 2.5 µl of Neuro-FECT transfection reagent (T800075; Genlantis, San Diego, CA, USA).
Behavioral tests were conducted at the indicated time points. After the behavioral tests, the injection sites were confirmed by a histological analysis, and the mice with the wrong injection site were excluded from the final data as described [20 (link)30 (link)31 (link)]. The following siRNAs were used: control-siRNA (SN-1012) and Trk-BsiRNA (1393919, NM_001025074.2) were purchased from Bioneer Co. (Deajun, Korea), and FAM-labeled RISK-independent siRNA transfection control siGLO Green (D-001630-01-05) was purchased from Dharmacon Inc. (Chicago, IL, USA).

Lee Y, & Han P.L. (2019). Early-Life Stress in D2 Heterozygous Mice Promotes Autistic-like Behaviors through the Downregulation of the BDNF-TrkB Pathway in the Dorsal Striatum. Experimental Neurobiology, 28(3), 337-351.

+ Open protocol

+ Expand

Stereotaxic siRNA Silencing in Mouse Hippocampus

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

Stereotaxic injection of siRNA was carried out as previously described^{38 (link)}. Mice were anesthetized by intraperitoneal injection of a mixture (3.5:1) of ketamine hydrochloride (50 mg/ml) and xylazine hydrochloride (23.3 mg/ml) at a dose of 2.5 μl/g body weight. siRNA-control (SN-1012), siRNA-Ppp2ca (#1411897, NM_019411.4), and siRNA-SUV39H1 (#1433203, NM_011514.1) were purchased from Bionner Co. (Daejeon, Korea). One volume of each siRNA (50 ng/μl) was mixed with 2.5 volumes of Neurofect transfection reagent (T800075; Genlantis, San Diego, CA, USA) and 0.5 volume of 50% sucrose. The siRNA mix was incubated for 20 min prior to stereotaxic injection. A volume of 1.8 μl of the siRNA mix (7.5 ng/μl) was injected into each CA3 region (stereotaxic coordinate: AP, −1.9; ML, ± 2.1; DV,−2.1 mm) at a speed of 0.2 μl/min using a stereotaxic injection system (Vernier Stereotaxic Instrument, Leica Biosystems, Wetzlar, Germany) and a Hamilton syringe with a 30 G needle. After injection, mice were kept on a warm pad until they were awakened, and housed afterward in home cages. Behavioral tests were performed between 48 h and 72 h after siRNA injection. The subjects were sacrificed for tissue preparation at the indicated time point.

Lee J.E., Kwon H.J., Choi J., Seo J.S, & Han P.L. (2020). Aging increases vulnerability to stress-induced depression via upregulation of NADPH oxidase in mice. Communications Biology, 3, 292.

+ Open protocol

+ Expand

Stereotaxic siRNA Injection for Targeted Gene Silencing

Cited 1 time

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

Stereotaxic injection of siRNA was performed as described previously [22 (link), 32 (link)]. In brief, mice were anesthetized with a mixture (3.5:1) of ketamine hydrochloride (50 mg/ml) and xylazine hydrochloride (23.3 mg/ml) at a dose of 2.5 μl/g body weight. One portion of diluted siRNA (50 ng/μl) was mixed with 2.5 portions Neurofect transfection reagent (T800075, Genlantis, San Diego, CA, USA), and 0.5 portion of 50% sucrose 20 min prior to injection and incubated on ice.
The siRNA mix (each, 1.8 μl of 7.5 ng/μl) was injected into the CA3 region (stereotaxic coordinate: AP, -1.9; ML, ±3.0; DV, -2.1 mm) on both sides using a stereotaxic injection system (Vernier Stereotaxic Instrument, Leica Biosystems, Wentzler, Germany) and a Hamilton syringe with a 30-G needle. After 5 min, the needle was removed in three intermediate steps for 3 min each. Mice were kept on a warm pad until awakened. Tissue samples with siRNA injection were prepared 48 h after injection.
Control siRNA (siCON, SN-1012) and MeCP2-siRNA (siMeCP2, 1385135; NM_001081979.2) were purchased from Bioneer Co. (Daejeon, Korea). The siRNAs were resolved to 50 ng/μl in a siRNA dilution buffer (B-002000-UB-100, Dharmacon, Lafayette, CO, USA).

Choi J., Kwon H, & Han P.L. (2021). Hyperoxygenation Treatment Reduces Beta-amyloid Deposition via MeCP2-dependent Upregulation of MMP-2 and MMP-9 in the Hippocampus of Tg-APP/PS1 Mice. Experimental Neurobiology, 30(4), 294-307.

+ 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!