For the depletion of UCHL1 and HIF‐1α in MDA‐MB‐436 cells, cells were plated in 6‐well or 24‐well tissue culture plates (Corning) at a concentration of 1.2 × 105 cells/mL and cultured in antibiotic‐free DMEM medium containing 10% of FBS. For transfection of 6‐well or 24‐well cultures, 10 or 50 pmol of siRNA (siUCHL1: silencer Select Validated siRNA, Cat# 4390824‐s14616; Thermo Fisher Scientific: 5′‐AAGUUAGUCCUAAAGUGUATT‐3′, 4390824‐s14617: 5′‐GCACAAUCGGACUUAUUCATT‐3′ siHIF‐1α: silencer Select Validated siRNA, Cat# 4390824‐s6539 and Cat# 4390824‐s6541) with Lipofectamine RNAiMAX or Lipofectamine 3000 (Thermo Fisher Scientific) was used, respectively. After 48‐hour transfection, the cells were lysed and subjected to western blotting analysis or immunofluorescent staining. For the negative control, scrambled siRNA (Cat# 12935‐300, sequence information not disclosed; Life Technologies) was used. For 3D culture, cells were first cultured and transfected with siRNA as indicated above, then collected after a 24‐hour incubation and cultured on 3D culture 96‐well plates (Corning).
Validated select sirna
Manufactured by Thermo Fisher Scientific
Validated select siRNA is a laboratory tool designed to facilitate gene silencing experiments. It provides pre-designed and validated small interfering RNA (siRNA) sequences targeting specific genes of interest. The core function of this product is to enable selective and efficient knockdown of target gene expression in cell-based assays, supporting research and investigations in various fields of study.
Automatically generated - may contain errors
Lab products found in correlation
Bmpr2, by Thermo Fisher Scientific (3 mentions)
Lipofectamine rnaimax reagent, by Thermo Fisher Scientific (3 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Scrambled sirna, by Thermo Fisher Scientific (1 mentions)
4 protocols using validated select sirna
1
Depletion of UCHL1 and HIF-1α in MDA-MB-436 Cells
2
BMPR2 Knockdown via siRNA
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Validated select siRNA was used to knockdown BMPR2 (Life Technologies). The ID numbers for the siRNA are: BMPR2 (s2044 and s2045). Silencer Select negative control siRNA (4390843) was used to evaluate selectivity. Transfections of the siRNA were performed in duplicate using Lipofectamine® RNAiMAX Reagent (Invitrogen, Carlsbad, CA, USA) according to manufacturer's protocol. Cells were transfected with 6 nM BMPR2 or 6 nM of siRNA control. The experiment was replicated three times in our laboratory.
3
BMPR2 Knockdown via siRNA
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Validated select siRNA was used to knockdown BMPR2 (Life Technologies). The ID numbers for the siRNA are: BMPR2 (s2044 and s2045). Silencer Select negative control siRNA (4390843) was used to evaluate selectivity. Transfections of the siRNA were performed in duplicate using Lipofectamine® RNAiMAX Reagent (Invitrogen, Carlsbad, CA, USA) according to manufacturer's protocol. Cells were transfected with 6 nM BMPR2 or 6 nM of siRNA control. The experiment was replicated three times in our laboratory.
4
BMPR2 Knockdown via siRNA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Validated select siRNA was used to knockdown BMPR2 (Life Technologies). The ID numbers for the siRNA are: BMPR2 (s2044 and s2045). Silencer Select negative control siRNA (4390843) was used to evaluate selectivity. Transfections of the siRNA were performed in duplicate using Lipofectamine® RNAiMAX Reagent (Invitrogen, Carlsbad, CA, USA) according to manufacturer's protocol. Cells were transfected with 6 nM BMPR2 or 6 nM of siRNA control. The experiment was replicated three times in our laboratory.
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