The following reagents were used: U0126 (Promega), epidermal growth factor (EGF; Millipore), mimosine (Sigma), rhTRAIL (Millipore), rhTNFα (Life Tech), TriplePrep Kit (GE-Healthcare). ON-TARGET plus SMARTpools siRNAs, NC (non-coding negative control oligos) and individual oligos constituting the pools were purchased from Dharmacon. RNAiMax and Lipofectamine LTX transfection reagents were from Invitrogen. Antibodies: αR-TRIB1 (Millipore), αR-CCND1, αM-CCNA2, αR-CDC25A, αR-IER2, αR-pCDK2, αR-pIKKa (Santa Cruz Biotechnology), αM-FLIP (Enzo Life Sciences), αM-BID (BD Biosciences), αM-DR5 (R&D Systems), all other antibodies were from Cell Signaling Technologies.
On target plus smartpools sirnas
Manufactured by Horizon Discovery
ON-TARGET plus SMARTpools siRNAs are a collection of predesigned, pooled small interfering RNA (siRNA) reagents designed to target specific gene transcripts. The core function of these siRNAs is to induce gene knockdown by triggering the RNA interference (RNAi) pathway within cells.
Automatically generated - may contain errors
Lab products found in correlation
Tripleprep kit, by GE Healthcare (1 mentions)
Rhtnf α, by Thermo Fisher Scientific (1 mentions)
Lipofectamine ltx, by Thermo Fisher Scientific (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Rnaimax, by Thermo Fisher Scientific (1 mentions)
Mimosine, by Merck Group (1 mentions)
U0126, by Promega (1 mentions)
On targetplus non targeting pool sirna, by Horizon Discovery (1 mentions)
Dharmafect 1, by Horizon Discovery (1 mentions)
2 protocols using on target plus smartpools sirnas
1
Investigating Cellular Signaling Pathways
2
Knockdown of KAT2A/GCN5 and KAT2B/PCAF in U2OS Cells
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For KAT2A/GCN5 and KAT2B/PCAF knockdowns, U2OS cells at 30% confluence were transfected using DharmaFECT 1 (Dharmacon) according to the manufacturer’s instructions, with 50 pmol each of ON-Targetplus SMARTpools siRNAs targeting KAT2A (Dharmacon L-009722-02-0005) and KAT2B (Dharmacon L-005055-00-0005) in serum-free DMEM. As a negative control, 100 pmol of ON-TARGETplus non-targeting pool siRNAs (Dharmacon D001810-10-05) were used. The serum-free medium was replaced with DMEM supplemented with 10% FBS at 24 hr after transfection, and after further 48 hr incubation, the cells were collected by trypsinisation (total of 72 hr siRNA exposure).
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