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Sc 45469

Manufactured by Santa Cruz Biotechnology

Sc-45469 is a piece of lab equipment manufactured by Santa Cruz Biotechnology. It is designed for basic laboratory procedures. The core function of Sc-45469 is to facilitate sample preparation and analysis. Further details about its intended use are not available.

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2 protocols using sc 45469

1

Silencing NLRP3 to Investigate IL-22 Effects

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To reduce endogenous NLRP3 expression, siRNA oligonucleotides (80 pmol, sc-45469; Santa Cruz) was used according to the manufacturer’s instructions. The cells were transfected with either NLRP3 siRNA oligonucleotide or a non-targeted control siRNA oligonucleotide and incubated for 6h at 37°C under standard culture conditions. Then the cells were cultured with fresh culture media for further 14 h. In the next moment, the cells were treated with 20 ng/ml of IL-22 for another 24h. Thereafter, the cell lysates were harvested for immunoblotting, the supernatants for Elisa assay.
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2

siRNA Knockdown of NLRP3, EP2, EP4, and cPLA2α

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ON-TARGETplus SMART pool small interfering RNA (siRNA) (Dharmacon, Thermo Scientific, Lafayette, CO) against human NLRP3 (L-017367-00-0005), or sc-45469, Santa Cruz), EP2 (PTGER2) (L-005712-00-0005), EP4 (PTGER4) (L-005714-00-0005) and cPLA2α (PLA2G4A) (L-009886-00-005) together with ON-TARGETplus Control Non-targeting pool (D-00181970-05) were used to perform knockdown experiments in human primary MDM or THP-1 cells. Human primary MDM (0.5×106 cells per 100 μl cuvette) or THP-1 cells (5×106 per 100 μl cuvette) were transfected with siRNA pools (1 μM or 100 nM) using a P3 Primary Cell 4D-Nucleofector X Kit L (Amaxa, Cologne, Germany) according to the manufacturer's protocols for human macrophages and monocytes, respectively. The silencing of gene expression was confirmed by RT-PCR. All experiments on transfected cells were performed after 48 hours.
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