Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
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MID1-3 siRNA: 5′-GUGUGAUACUAGGAUGCGG-3′
siRNA final concentration- 20 to 40 nM
Add diluted siRNA to diluted Lipofectamine Reagent
Incubate for 5 min at RT and add to cells.
Incubate cells for 1–3 days at 37°C. |
|
Protocol tips |
MID1-3 siRNA: 5′-GUGUGAUACUAGGAUGCGG-3′
siRNA final concentration- 20 to 40 nM
Add diluted siRNA to diluted Lipofectamine Reagent
Incubate for 5 min at RT and add to cells.
Incubate cells for 1–3 days at 37°C. |
Publication protocol
Cells were transfected with 20 to 40 nM final concentration of MID1, α4, AR or control siRNA oligonucleotides using Lipofectamin 2000 or Nanofectin. AR siRNA: 5′-GAAAGCUCCUCGGUAGGUC-3′; α4-4 siRNA: 5′-UUGAGAUGCCAUAGCAACGAG-3′; MID1-3 siRNA: 5′-GUGUGAUACUAGGAUGCGG-3′; ns siRNA: 5′-AAGAGGCUUGCACAGUGCA-3′ (Dharmacon or MWG). After 48 h of incubation the medium was replaced and after 72–120 h cells were harvested and analyzed by Western blotting or real-time PCR
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Papers
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Paper title
A hormone-dependent feedback-loop controls androgen receptor levels by limiting MID1, a novel translation enhancer and promoter of oncogenic signaling
Manufacturer protocol
Download the product protocol from Dharmacon for ON-TARGETplus Human MID1 (4281) siRNA - Individual below.
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