Monocytes were washed and resuspended in 100 µl of nucleofector solution provided with the Amaxa Noclecfector Kit V and transfected separately with siRNA-n-SMase (Santa Cruz Biotechnology, INK. SC-106277), scramble (control) siRNA (30 nM; OriGene Technologies, Inc. MD, USA, USA), and pmaxGFP (0.5 ug; Amaxa Nucleofector Kit V for THP-1, Lonza). All transfection experiments were performed with Amaxa Cell Line Nucleofector Kit V for monocytic cells(Lonza, Germany) by using Amaxa Electroporation System (Amaxa Inc, Germany) according to the manufacturer's protocol37 (link). After 36 h of transfection, cells were treated with TNF-α for 2 h. For knock down of nSMase2 in human primary monocyte cultures, cells were transfected with 20 nM of the siRNA using Viromer Blue (lipocalyx, Halle, Germany) according to the manufacturer’s instructions. Cells were harvested for RNA isolation and staining study. nSMase gene knock down level was assessed by Real Time-PCR using SMPD3 gene-specific primer probes.
Sirna n smase
Manufactured by Santa Cruz Biotechnology
Sourced in United States
SiRNA-n-SMase is a laboratory product designed for the study of sphingomyelinase (SMase) gene expression. It contains a small interfering RNA (siRNA) molecule that targets the mRNA of the neutral sphingomyelinase (n-SMase) gene, allowing for the modulation of n-SMase expression in experimental settings.
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Lab products found in correlation
2 protocols using sirna n smase
1
Monocyte Transfection and nSMase2 Knockdown
2
Monocyte Transfection and nSMase2 Knockdown
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Monocytes were washed and resuspended in 100 ul of nucleofector solution provided with the Amaxa Noclecfector Kit V and transfected separately with siRNA-n-SMase (Santa Cruz Biotechnology, INK. SC-106277), scramble (control) siRNA (30nM; OriGene Technologies, Inc. MD, USA, USA), and pmaxGFP (0.5 ug; Amaxa Noclecfector Kit V for THP-1, Lonza). All transfection experiments were performed with Amaxa Cell Line Nucleofector Kit V for monocytic cells(Lonza, Germany) by using Amaxa
Electroporation System (Amaxa Inc, Germany) according to the manufacturer's protocol (13) . After 36 hours of transfection, cells were treated with TNF-α for 2 hours. For knock down of nSMase2 in human primary monocyte cultures, cells were transfected with 20 nM of the siRNA using Viromer Blue (lipocalyx, Halle, Germany) according to the manufacturer's instructions. Cells were harvested for RNA isolation and staining study. nSMase gene knock down level was assessed by Real Time-PCR using SMPD3 genespecific primer probes.
Electroporation System (Amaxa Inc, Germany) according to the manufacturer's protocol (13) . After 36 hours of transfection, cells were treated with TNF-α for 2 hours. For knock down of nSMase2 in human primary monocyte cultures, cells were transfected with 20 nM of the siRNA using Viromer Blue (lipocalyx, Halle, Germany) according to the manufacturer's instructions. Cells were harvested for RNA isolation and staining study. nSMase gene knock down level was assessed by Real Time-PCR using SMPD3 genespecific primer probes.
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