Human telomerase immortalized retinal pigment epithelium cells (hTERT-RPE1, ATCC) were cultured with Dulbecco’s modified Eagle’s Medium DMEM/F12 50/50 medium (Pan Biotech) supplemented with 10% fetal bovine serum (FBS, Life Technologies) and1% penicillin-streptomycin (Gibco). Human embryonic kidney (HEK293T, ATCC) and osteosarcoma epithelial (U2OS, ATCC) cells were cultured with DMEM medium (Pan Biotech) supplemented with10% FBS and 1% penicillin-streptomycin. All cell lines were authenticated by Multiplex Cell Line Authentication (MCA) and were tested for mycoplasma by MycoAlert Mycoplasma Detection Kit (Lonza). U2OS cells were transfected with the plasmids using Lipofectamine 2000 and according to the manufacturer’s instructions (Thermo Scientific). HEK293T cells were transfected with the plasmids using 1 mg/ml polyethylenimine, MW 25 kDa (PEI). For microtubule depolymerization experiments, cells were treated with 5 μg/ml nocodazole (Sigma-Aldrich) or vehicle (dimethyl sulfoxide) for 1 h at 37°C. For cell synchronization experiments, 5 μm (+)-S-trityl-L-cysteine (STLC) (Alfa-Aesar) was used for 16 h at 37°C.
Dmem f12 50 50 medium
Manufactured by PAN Biotech
DMEM/F12 50/50 medium is a cell culture medium used in laboratory settings. It is a basal medium that provides the necessary nutrients and components to support the growth and maintenance of a variety of cell types. The medium is a 1:1 mixture of Dulbecco's Modified Eagle's Medium (DMEM) and Ham's F-12 Nutrient Mixture, which combines the formulations of these two widely used media.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Nocodazole, by Merck Group (2 mentions)
S trityl l cysteine, by Thermo Fisher Scientific (2 mentions)
Mycoalert mycoplasma detection kit, by Lonza (2 mentions)
Dmem medium, by PAN Biotech (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Htert rpe1, by American Type Culture Collection (1 mentions)
Penicillin streptomycin, by PAN Biotech (1 mentions)
Fetal bovine serum (fbs), by PAN Biotech (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
2 protocols using dmem f12 50 50 medium
1
Cultivation and Transfection of RPE1, HEK293T, and U2OS Cells
2
Immortalized Retinal Epithelial Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human telomerase immortalized retinal pigment epithelium cells (hTERT-RPE1, ATCC, CRL-4000) were cultured with Dulbecco's modified Eagle's Medium DMEM/F12 50/50 medium (Pan Biotech,Cat. # P04-41250) supplemented with 10% Fetal Bovine Serum (FBS, Life Technologies, Ref. # 10270-106, Lot # 42Q5283K) and1% penicillin-streptomycin (GIBCO, Cat. # 1540-122). Human embryonic kidney (HEK293T, ATCC, CRL-3216), and osteosarcoma epithelial (U2OS, ATCC, HTB-96) cells were cultured with DMEM medium (Pan Biotech, Cat. # P04-03590) supplemented with10% FBS and 1% penicillin-streptomycin. All cell lines were authenticated by Multiplex Cell Line Authentication (MCA) and were tested for mycoplasma by MycoAlert Mycoplasma Detection Kit (Lonza). U2OS cells were transfected with the plasmids using Lipofectamine 2000 and according to the manufacturer's instructions (Thermo Scientific Scientific). HEK293T cells were transfected with the plasmids using 1mg/ml polyethylenimine, MW 25 kDa (PEI). For microtubule depolymerization experiments, cells were treated with 5 µg/ml nocodazole (Sigma-Aldrich, Cat. #M1404) or vehicle (dimethyl sulfoxide) for one hour at 37 C. For cell synchronization experiments, 5 µM (+)-Strityl-L-cysteine (STLC) (Alfa-Aesar, Cat. #2799-07-7) was used for 16 h at 37 C.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!