ON-TARGETplus Rat Becn1 (114558) siRNA - SMARTpool

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	The rat UMR-106 osteosarcoma cell line75 was maintained in Dulbecco modified Eagle's medium (Lonza, BE12-604) supplemented with 10% Hyclone fetal calf serum (Thermo Scientific, SH30071.03). The UMR-106 GFP-LC3 clone 4 cells correspond to the UMR-106 cell line stably transfected with the pGFP-LC3 construct. These cells are maintained in the same medium in the presence of G418 (800 μg/ml; Sigma-Aldrich, G8168). For mineralization, the cells were cultured in α-MEM (Lonza, BE02-002) supplemented with 10% Hyclone fetal calf serum, CaCl2 (1.4 mM; Merck 2382), ascorbic acid (50 μg/mL; Sigma-Aldrich A4034) and dexamethasone (20 μg/mL; Sigma-Aldrich, D8893) for 3 d, and then for 3 d in the same medium in the presence of β-glycerophosphate (50 mg/mL; Sigma-Aldrich G9891). ON-TARGET plus SMARTpool siRNA (Dharmacon RNAi Technologies, Abgene Ltd) for rat Atg7 (L095596-01) and Becn1 (L-099237-01) were transfected in the UMR-106 cell line using nucleofection and Amaxa solution V (Lonza, VCA-1003).

Protocol tips

The rat UMR-106 osteosarcoma cell line75 was maintained in Dulbecco modified Eagle's medium (Lonza, BE12-604) supplemented with 10% Hyclone fetal calf serum (Thermo Scientific, SH30071.03). The UMR-106 GFP-LC3 clone 4 cells correspond to the UMR-106 cell line stably transfected with the pGFP-LC3 construct. These cells are maintained in the same medium in the presence of G418 (800 μg/ml; Sigma-Aldrich, G8168). For mineralization, the cells were cultured in α-MEM (Lonza, BE02-002) supplemented with 10% Hyclone fetal calf serum, CaCl2 (1.4 mM; Merck 2382), ascorbic acid (50 μg/mL; Sigma-Aldrich A4034) and dexamethasone (20 μg/mL; Sigma-Aldrich, D8893) for 3 d, and then for 3 d in the same medium in the presence of β-glycerophosphate (50 mg/mL; Sigma-Aldrich G9891). ON-TARGET plus SMARTpool siRNA (Dharmacon RNAi Technologies, Abgene Ltd) for rat Atg7 (L095596-01) and Becn1 (L-099237-01) were transfected in the UMR-106 cell line using nucleofection and Amaxa solution V (Lonza, VCA-1003).

Publication protocol

The rat UMR-106 osteosarcoma cell line75 was maintained in Dulbecco modified Eagle's medium (Lonza, BE12-604) supplemented with 10% Hyclone fetal calf serum (Thermo Scientific, SH30071.03). The UMR-106 GFP-LC3 clone 4 cells correspond to the UMR-106 cell line stably transfected with the pGFP-LC3 construct. These cells are maintained in the same medium in the presence of G418 (800 μg/ml; Sigma-Aldrich, G8168). For mineralization, the cells were cultured in α-MEM (Lonza, BE02-002) supplemented with 10% Hyclone fetal calf serum, CaCl2 (1.4 mM; Merck 2382), ascorbic acid (50 μg/mL; Sigma-Aldrich A4034) and dexamethasone (20 μg/mL; Sigma-Aldrich, D8893) for 3 d, and then for 3 d in the same medium in the presence of β-glycerophosphate (50 mg/mL; Sigma-Aldrich G9891). ON-TARGET plus SMARTpool siRNA (Dharmacon RNAi Technologies, Abgene Ltd) for rat Atg7 (L095596-01) and Becn1 (L-099237-01) were transfected in the UMR-106 cell line using nucleofection and Amaxa solution V (Lonza, VCA-1003).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing siRNA / miRNA gene silencing Rat - UMR‐106 Becn1 using ON-TARGETplus Rat Becn1 (114558) siRNA - SMARTpool from Horizon Discovery Ltd..

Manufacturer protocol

Download the product protocol from Horizon Discovery Ltd. for ON-TARGETplus Rat Becn1 (114558) siRNA - SMARTpool below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing siRNA / miRNA gene silencing Rat - UMR‐106 Becn1 using ON-TARGETplus Rat Becn1 (114558) siRNA - SMARTpool from Horizon Discovery Ltd.. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.