hUC-MSCs were transfected with 3 μg/mL of either the pcDNA3.1/CT-GFP-Lcn2 (Lcn2-MSCs) or the pcDNA3.1/CT-GFP empty vector (V-MSCs) using X-tremeGENE HP DNA transfection kit (Roche, Germany) according to the manufacturers’ protocol and as described previously.31 (link)
X tremegene hp dna transfection kit
Manufactured by Roche
Sourced in Germany
The X-tremeGENE HP DNA Transfection Kit is a laboratory product designed for the efficient transfection of a wide range of cell types with DNA. It provides a simple and effective method for introducing genetic material into cells.
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Lab products found in correlation
4 protocols using x tremegene hp dna transfection kit
1
Lcn2 Overexpression in hUC-MSCs
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hUC-MSCs were transfected with 3 μg/mL of either the pcDNA3.1/CT-GFP-Lcn2 (Lcn2-MSCs) or the pcDNA3.1/CT-GFP empty vector (V-MSCs) using X-tremeGENE HP DNA transfection kit (Roche, Germany) according to the manufacturers’ protocol and as described previously.31 (link)
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hUC-MSCs were transfected with 3 μg/mL of either the pcDNA3.1/CT-GFP-Lcn2 (Lcn2-MSCs) or the pcDNA3.1/CT-GFP empty vector (V-MSCs) using X-tremeGENE HP DNA transfection kit (Roche, Germany) according to the manufacturers’ protocol and as described previously.31 (link)
2
Optimizing KYSE30 Cell Transfection with X-treme GENE HP
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To optimize transfection with X-treme GENE HP
DNA transfection kit (Roche. Germany), freshly KYSE30
cells were first passaged approximately 24 hr before
transfection. Optimal cell density was 70 to 90%
confluence for KYSE30 transfection. We used 1:1, 2:1,
3:1 and 4:1 ratios of micro liter (µl) X-treme GENE HP
DNA Transfection Reagent to microgram (µg) pB-GFP
vector as control for transfection. According to
manufacturer's protocols, DNA was diluted with serumfree
medium (RPMI 1640) (PAA, Australia) to a final
concentration of 1 µg plasmid DNA per 100 µl medium
(0.01 µg/µl), and X-treme GENE HP DNA transfection
reagent was added directly into the medium containing
the diluted DNA, and incubated for 15 min at +15 to
+25°C, then the mix was added to the KYSE30 cells.
Finally, the transfected cells were selected using the
treatment with G418 antibiotic after 5 days.
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To optimize transfection with X-treme GENE HP
DNA transfection kit (Roche. Germany), freshly KYSE30
cells were first passaged approximately 24 hr before
transfection. Optimal cell density was 70 to 90%
confluence for KYSE30 transfection. We used 1:1, 2:1,
3:1 and 4:1 ratios of micro liter (µl) X-treme GENE HP
DNA Transfection Reagent to microgram (µg) pB-GFP
vector as control for transfection. According to
manufacturer's protocols, DNA was diluted with serumfree
medium (RPMI 1640) (PAA, Australia) to a final
concentration of 1 µg plasmid DNA per 100 µl medium
(0.01 µg/µl), and X-treme GENE HP DNA transfection
reagent was added directly into the medium containing
the diluted DNA, and incubated for 15 min at +15 to
+25°C, then the mix was added to the KYSE30 cells.
Finally, the transfected cells were selected using the
treatment with G418 antibiotic after 5 days.
3
Plasmid Transfection of AML Cells
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AML cells were inoculated into 6-well cell culture plates at a concentration of 5 × 105 /mL per well. When the cells growth density reached more than 70%, plasmids were transfected using X-tremeGENE HP DNA transfection kit (Roche) according to the manufacturer’s instructions [37 (link)].
4
Hippocampal Neuron Culture and Transfection
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Sprague–Dawley pregnant rats (Janvier Labs) were killed according to the European Directive rules (2010/63/EU). Dissociated hippocampal neurons from E18 embryos were prepared as described previously (Kaech and Banker, 2006 (link)) at a density of 300,000 cells per 60 mm dish on poly-L-lysine pre-coated coverslips. Neurons were transfected with the cDNA using Ca2+ method at 8–11 days in vitro. Proteins of interest are expressed for 3–6 days depending on the experiments. For cLTP experiments expression is started two days before the experiments by the addition of 500 nM of doxycycline in the media. All experiments were performed in accordance with the European guidelines for the care and use of laboratory animals, and the guidelines issued by the University of Bordeaux animal experimental committee (CE50; Animal facilities authorizations A3306940, A33063941).
COS-7 cells were maintained in Dulbecco′s Modified Eagle′s Medium (DMEM 4,5 G/L+GLUT & PYRUVATE 500, Eurobio) with 10% fetal bovine serum (Eurobio) and 1% L-glutamine (Gibco). Transfection was done with the Xtreme gene HP DNA transfection kit (Roche) following the manufacturer’s protocol.
COS-7 cells were maintained in Dulbecco′s Modified Eagle′s Medium (DMEM 4,5 G/L+GLUT & PYRUVATE 500, Eurobio) with 10% fetal bovine serum (Eurobio) and 1% L-glutamine (Gibco). Transfection was done with the Xtreme gene HP DNA transfection kit (Roche) following the manufacturer’s protocol.
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