Human SH-SY5Y neuroblastoma and human embryonic kidney cells (HEK293) were grown at 37°C in a humidified incubator chamber under an atmosphere of 7.5% CO2 in DMEM supplemented with 10% (v/v) heat-inactivated FCS, 2 mM Glutamax, and 1% (v/v) penicillin/streptomycin. Cells were passaged 1–2 times per week, and plated for treatment when they reached 80–90% confluence. SH-SY5Y cells were stably transfected with DNA constructs harboring human wild-type APP695 (APP) or the expression vector pCEP4 (Invitrogen, Saint Aubin, France) alone (control vector, Co) [34 (link)]. Transfected APP cells were grown in DMEM standard medium supplemented with 300 μg/ml hygromycin. HEK cells overexpressing Swedish APP (HEK SWE APP) cells were stably transfected with DNA constructs harboring human wild-type APP695 (APP) and were grown in DMEM standard medium supplemented with 300 μg/ml G418.
Control vector
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Control vector is a plasmid DNA construct that serves as a reference standard in molecular biology experiments. It contains essential genetic elements for DNA propagation and expression, allowing for the verification of experimental procedures and the evaluation of target gene expression levels.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (5 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (2 mentions)
Pcdna β catenin, by Addgene (2 mentions)
Pcdna β catenin, by Thermo Fisher Scientific (2 mentions)
β catenin sirna, by Thermo Fisher Scientific (2 mentions)
Control sirna, by Thermo Fisher Scientific (2 mentions)
Pcep4, by Thermo Fisher Scientific (1 mentions)
Lacz vector, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000 kit, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Sartorius (1 mentions)
Streptomycin, by Sartorius (1 mentions)
Cell incubator, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Control sirna sc 37007, by Santa Cruz Biotechnology (1 mentions)
Bovine pituitary extract, by Thermo Fisher Scientific (1 mentions)
Human recombinant epidermal growth factor, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
9 protocols using control vector
1
Stable Transfection of APP in Neuroblastoma and Kidney Cells
2
Sirt1 Modulation of Astrocyte Activation
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To evaluate the potential effect of Sirt1 on activation of astrocytes in vitro, the astrocytes were transfected with control or Sirt1 for 24 h and then stimulated with 1 ng/ml IL-1β. The Sirt1-recombined adenoviral expression vector and the control vector were constructed (Invitrogen, Carlsbad, CA, USA) and performed at a multiplicity of infection (MOI) of 20 [35 (link), 36 (link)]. The lacZ vector (Invitrogen) was added to control groups.
Cells (1 × 106) grown to 60–80% confluence in 24 wells were transfected with adenoviral vectors using a Lipofectamine 2000 kit (Invitrogen, cat. 11668–019) according to the procedure provided by the manufacturer. The cells were observed under a fluorescence microscope and harvested 24 h after transfection.
Cells (1 × 106) grown to 60–80% confluence in 24 wells were transfected with adenoviral vectors using a Lipofectamine 2000 kit (Invitrogen, cat. 11668–019) according to the procedure provided by the manufacturer. The cells were observed under a fluorescence microscope and harvested 24 h after transfection.
3
Transfection of SW620 cells with Wnt 5 and β-catenin
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SW620 cell line was purchased from ATCC. SW620 cells were cultured with DMEM (#06-1170-87-1A, Biological Industries, Israel) containing 10% fetal bovine serum (#04-011-1A, Biological Industries, Israel), 100 U/ml penicillin and 100 mg/mL streptomycin (#03-034-1B, Biological Industries, Israel). SW620 cells were incubated in a cell incubator (Thermo Scientific, USA) containing 5% CO2, at 37 °C.
Wnt 5 (#sc-41112) and β-catenin (#sc-29209) siRNA and control (#sc-37007) siRNA were purchased from Santa Cruz Biotechnology (CA, USA). pcDNA-Wnt 5 and pcDNA-β-catenin and control vector were purchased from Addgene (Cambridge, UK). SW620 cells were cultured in DMEM medium at a density of 105 cells in 6-well plates. According to the manufacturer's instructions, lipofectamine 3000 reagent (#L3000015, Invitrogen, CA, USA), pcDNA-Wnt 5 (2 μg), pcDNA-β-catenin (2 μg), control vector (2 μg) or wnt 5 siRNA, β-catenin siRNA (50 nM) and control siRNA (50 nM) were used to transiently transfect into cells. After 6 h of transfection, complete culture medium was added to the transfection medium, continue culturing for 12 h. Then the cells are collected for inoculation.
Wnt 5 (#sc-41112) and β-catenin (#sc-29209) siRNA and control (#sc-37007) siRNA were purchased from Santa Cruz Biotechnology (CA, USA). pcDNA-Wnt 5 and pcDNA-β-catenin and control vector were purchased from Addgene (Cambridge, UK). SW620 cells were cultured in DMEM medium at a density of 105 cells in 6-well plates. According to the manufacturer's instructions, lipofectamine 3000 reagent (#L3000015, Invitrogen, CA, USA), pcDNA-Wnt 5 (2 μg), pcDNA-β-catenin (2 μg), control vector (2 μg) or wnt 5 siRNA, β-catenin siRNA (50 nM) and control siRNA (50 nM) were used to transiently transfect into cells. After 6 h of transfection, complete culture medium was added to the transfection medium, continue culturing for 12 h. Then the cells are collected for inoculation.
4
Establishing RCC Cell Line Cultures
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Five RCC cell lines (ACHN, 786-O, SN12PM6, OS-RC-2 and CAKI-2) and human proximal tubule epithelial cell line HK-2 cell line were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). RCC cells were cultured in RPMI 1640 medium (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS, Gibco), 100 units/mL penicillin and 100 μg/mL streptomycin (Sigma, St-Louis, MO, USA). HK-2 cells were maintained in complete RPMI 1640 medium supplemented with keratinocyte serum-free medium, bovine pituitary extract, and human recombinant epidermal growth factor (Invitrogen, Carlsbad, CA, USA).
MiR-212 overexpressing vector, the control vector, miR-212 inhibitor and the negative control were bought from the company of Genecopoeia (Guangzhou, China). Plasmids expressing XIAP were purchased from Addgene (Cambridge, MA, USA). XIAP specific shRNAs were purchased from OriGene (Beijing, China). These vectors were transfected into RCC cells using Lipofectamine 2000 according to the manufacturer's instructions (Invitrogen, Carlsbad, CA, USA).
MiR-212 overexpressing vector, the control vector, miR-212 inhibitor and the negative control were bought from the company of Genecopoeia (Guangzhou, China). Plasmids expressing XIAP were purchased from Addgene (Cambridge, MA, USA). XIAP specific shRNAs were purchased from OriGene (Beijing, China). These vectors were transfected into RCC cells using Lipofectamine 2000 according to the manufacturer's instructions (Invitrogen, Carlsbad, CA, USA).
5
NEAT1 Knockdown and Overexpression in HepG2 Cells
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Small interfering RNAs (siRNAs) against NEAT1 (5′-UGGUAAUGGUGGAGGAAGAUU-3′, 5′-GUGAGAAGUUGCUUAGAAAUU-3′), a scrambled negative control siRNA (cat. no. siN05815122147-1-5), a miR-129-5p inhibitor (cat. no. miR20000242-1-5) and a corresponding negative control vector (cat. no. NC-miR20000242-1-5) were purchased from Guangzhou RiboBio Co., Ltd. (Guangzhou, China). A NEAT1-expressing vector and a control vector were purchased from Invitrogen (Thermo Fisher Scientific, Inc.). The aforementioned vectors were transfected into HepG2 cells with Lipofectamine 2000® (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions.
6
Modulating β-catenin in Lung Cancer Cells
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β-catenin siRNA and control siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). pcDNA-β-catenin and control vector were purchased from Addgene (Cambridge, MA, USA). A549 and H1299 cells were cultured in 6-well plates at a density of 105 cells in PRMI1640 medium. After incubation for 12 h, the cells were transiently transfected with pcDNA-β-catenin (2 μg), control vector (2 μg), or β-catenin siRNA (50 nM), and control siRNA (50 nM), using lipofectamine 3000 reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. After transfection for 4–6 h, cells were collected and plated in SFM for another 3 days with or without apatinib.
7
Evaluation of miR-146a Regulation of NOS1 3'UTR
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The 3′UTR of NOS1 genes were amplified by PCR, and the PCR products were inserted into the XhoI and XbaI sites of the pmirGlo plasmid (Promega, WI) according to standard protocol. The KOD-plus mutagenesis kit (Toyobo, Osaka, Japan) was used to construct the pmirGlo plasmids with mutated NOS1 3′UTR, in accordance with the manufacturer’s protocol. The mutagenesis was inserted into the same site of a control vector (Ambion, Cambridgeshire, UK) at the same time. Lipofectamine 2000 (Invitrogen, Life Technologies, CA) was used to transfect the PC-3 cells with miR-146a mimics (50 nM) and plasmid (5 ng/mL) in accordance with the manufacturer’s recommendation. The Dual-Glo luciferase assay kit (Promega, WI) was used to detect the luciferase activity based on the manufacturer’s instructions. All tests were performed 3 times.
8
MiR-300 Mimics: Cell Transfection
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MiR-300 mimics and scramble oligonucleotides (10nmol/l), ROCK1 vector and control vector were obtained from Ambion (Ambion, USA). Cell transfection was performed using Lipofectamine 2000 (Life Technologies, USA) according to manufacturer's information.
9
MiR-142 Mimics and HMGB1 Plasmid Transfection
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MiR-142 mimics (sense: 5′-UAGCAGCACAUCAU GGUUUACA-3′, antisense: 5′-UAAACAUGAUGUG CUGCUGUU-3′), control miRNAs (10nmol/l), HMGB1 plasmids and control vector were all obtained from Ambion (Ambion, USA). And then cell transfection was carried out using lipofectamine 2000 (Life Technologies, USA) according to manufacturer's instructions.
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