siRNA for Beclin-1 (Rat) was purchased from Cell Signaling Techology (CST, #6246). rPT cells were seeded in 6-well plates and transfected with 100 nmol/L Beclin-1 siRNA in 1 mL of medium containing Lipofectamine 3000 (Invitrogen) according to the manufacturer’s protocol, for 60 h prior to cell lysis.
Beclin1 sirna
Manufactured by Thermo Fisher Scientific
Sourced in United States
Beclin1 siRNA is a small interfering RNA (siRNA) designed to target the Beclin1 gene, which is involved in the regulation of autophagy, a cellular process that recycles damaged or unwanted components. This siRNA can be used for research purposes to study the role of Beclin1 in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Opti mem, by Thermo Fisher Scientific (2 mentions)
Rad001, by Merck Group (1 mentions)
Mouse anti β actin antibody, by Merck Group (1 mentions)
Rabbit anti cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Annexin 5 apoptosis detection kit, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
Matrigel, by BD (1 mentions)
Beclin1 sirna, by GenePharma (1 mentions)
Opti mem 1 medium, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Sirnas, by Merck Group (1 mentions)
Non specific scrambled sirna, by Merck Group (1 mentions)
Edta free trypsin, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Lipofectamine rnaimax reagent, by Thermo Fisher Scientific (1 mentions)
5 protocols using beclin1 sirna
1
Beclin-1 siRNA Knockdown in Rat Cells
2
Targeting Beclin1 Modulates Autophagy in Cancer Cells
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LNCaP and PC3 cell lines were obtained from the American Type Culture Collection (ATCC); FBS, RPMI medium 1640, Dulbecco's modified Eagle's medium (DMEM), sodium pyruvate, L-glutamine, Penicillin, and Streptomycin were obtained from Hyclone; Rad001 and ATO were from Sigma; CellTiter assay was from Promega; Caspase-3/7 Assay Kit was from Anaspec; Annexin V apoptosis detection Kit was from eBioscience; Dharmafect transfection reagent was from Thermo Scientific Life Science; Lipofectamine 2000 transfection reagent was from Invitrogen; Beclin1-siRNA (5′-rGrGrArArUrGrGrArArUrGrArrGrArUrUrArA-3′, 5′-rArGrCr ArGrCrArUrUrArArUrCrUrCrArUrU-3′) and control-siRNA (5′-rGrArArArArArCrUrCrArUrArUrArArArUr Cr-3′, 5′-rGrUrGrGrGrGrCrGrA rUrUrUrArUrArUrGrA-3′) were from IDT; RT-PCR primers for Beclin1, 5′-GGCC AATAAGATGGGTCTGA-3′ and 5′-CTGCACACAG TCCAGGAAAG-3′; for GAPDH 5′-CATGGGTGTGA ACCATGAGA-3′ and 5′-CAGTGATGGCATGGACT GTG-3′, were from Valuegene. Rabbit anti-LC-3 polyclonal antibody was from GenScript; Rabbit anti-Beclin1, Rabbit anti-cleaved Caspase-3 and Rabbit anti-PARP1 polyclonal antibodies were from Cell Signaling; Rabbit anti-Atg5-Atg12 monoclonal antibody was from Epitomics; Mouse anti-β-Actin antibody was from Sigma; Matrigel was from BD Biosciences.
3
Beclin1 Knockdown and GFP-LC3 Assay
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Beclin1 siRNA (sense 5′-GGAGCCAUUUAUUGAAACUTT-3′ and antisense 5′-AGUUUCAAUAAAUGGCUCCTT-3′) was synthesized by Shanghai GenePharma Co., Ltd. (Shanghai, China). Transfection of cells with green fluorescent protein (GFP)-LC3 plasmid (provided by Dr. Ye Wei) or Beclin1 siRNA was carried out using Lipofectamine 2000 (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) according the manufacturer's protocol. Briefly, 4 µg GFP-LC3 plasmid, 100 pmol Beclin1 siRNA and 5 µl Lipofectamine 2000 were diluted in 50 µl Opti-MEM I medium (Invitrogen; Thermo Fisher Scientific, Inc.), respectively, and were incubated for 5 min at room temperature. The diluted GFP-LC3 plasmid and Beclin1 siRNA were mixed with Lipofectamine 2000, separately, and were incubated for 20 min at room temperature. The complexes were then added to 6-well plates, 2×105 cells/well for transfection. Cabazitaxel was added to the cells at 5 µg/ml/well 48 h post-transfection, and the cells were incubated for a further 24 h at 37°C. LC3 puncta was visualized by fluorescence microscopy, and the protein expression levels of Beclin1 were determined by western blotting.
4
Silencing Autophagy Genes in Osteosarcoma
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Small interfering RNAs (siRNAs) against ATG5 (sense: 5′-GUGAGAUAUGGUUUGAAUAdTdT-3′ and antisense: 3′-UAUUCAAACCAUAUCUCACdTdT-5′), Beclin 1 (sense: 5′-GUUUGGAGAUCUUAGAGCAdTdT-3′ and antisense: 3′-UGCUCUAAGAUCUCCAAACdTdT-5′) and a nonspecific scrambled siRNA were purchased from Sigma-Aldrich (Merck KGaA). HOS and U2OS cells (5 × 105) were seeded in six-well plates and transfected with Lipofectamine 3000 (Invitrogen, Rockville, MD, USA) mixed with a serum-free medium containing ATG5 siRNA, Beclin 1 siRNA, and scrambled siRNA. Transfection was performed under 5%CO2 at 37 °C for 24 h.
5
ESCC Cell Culture and Beclin1 Knockdown
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We used human KYSE-150 and KYSE30 cells (purchased from the Cell Bank of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) as the ESCC cell line, which were cultured in 10% FBS supplemented RPMI-1640 (Gibco) at 37°C in an incubator with 95% air and 5% carbon dioxide[27 (link)], The cell culture medium was replaced once every 2 days and treated with EDTA-free trypsin (Gibco) when the density of ESCC cells was 70–80%, The supernatant was obtained and centrifuged at 150 g for 5 min to collect cells. Cells were then seeded in a 6-well plate at a density of 1 × 105 cells per well and 70–80% confluency the next day. Lipofectamine RNAiMAX Reagent (Invitrogen) was diluted with Opti-MEM™ (Gibco) (dilution ratio1:16) and Beclin1 siRNA (Invitrogen) was diluted to 30 pmol with Opti-MEM™ according to the manufacturer’s instructions. Diluted siRNA was added to diluted Lipofectamine® RNAiMAX Reagent (dilution ratio1:1) and incubated for 5 min at room temperature. The siRNA-Lipofectamine complex was added to cells for 24 h, and the knockdown efficiency was detected by Western blotting.
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