U2OS cell lines were grown in DMEM (Invitrogen) supplemented with 10% calf serum (Atlanta Biologicals). Plasmid transfections were performed in OPTI-MEM media (Invitrogen) with Lipofectamine 2000 (Invitrogen). siRNA transfections used RNAmax (Invitrogen). Cells were analyzed 24 hr and 72–80 hr post-transfection for DNA and RNAi respectively. Cells were treated with MitoTracker Red CMXRos (Invitrogen) at 100 nM in DMEM 20 min prior to fixation. Chemical inhibitor treatments were 50 µM blebbistatin (Sigma-Aldrich), 200 µM CK666 or CK689 (Calbiochem), or 0.5 µM Latrunculin B (Calbiochem). Further details are in Supplementary Information .
Rnamax
Manufactured by Thermo Fisher Scientific
Sourced in United States
RNAmax is a high-performance RNA extraction kit designed for efficient and consistent isolation of total RNA from a variety of sample types, including cells, tissues, and biofluids. The kit utilizes a proprietary extraction method to ensure the preservation of RNA integrity and yield, making it a reliable tool for downstream applications such as gene expression analysis, RT-qPCR, and NGS.
Automatically generated - may contain errors
Lab products found in correlation
Opti mem media, by Thermo Fisher Scientific (1 mentions)
Calf serum, by Atlanta Biologicals (1 mentions)
Blebbistatin, by Merck Group (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Mitotracker red cmxros, by Thermo Fisher Scientific (1 mentions)
Evos m5000, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
2 protocols using rnamax
1
Cell Line Maintenance and Transfection
2
3D Co-culture of Glioblastoma and Microglia
Check if the same lab product or an alternative is used in the 5 most similar protocols
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U251 VCT/PDIA5 and U251 siNC/siPDIA5 were co-cultured with HMC3 GFP in 3D condition. In brief, PDIA5 over expression and Vector (VCT) plasmids were transfected via Lipofectamine 3000 (Invitrogen, US). Simultaneously. siNC and siPDIA5 RNA transfections were performed via RNA Max (Invitrogen, US). Two days post-transfection, tumor, and HMC3 GFP cells were genteelly digested and counted at 5x10’/each, then mixing in 200 µl organoids medium. U251 VCT/PDIA5-HMC3 GFP and U251 siNÇ/siPDIA5-HMC3 GFP in organoids medium were divided and planted 40 ul/droplet. Three days post-plantation, droplets were monitored and imaged by EVOS M5000 (Invitrogen, US). The second timepoint of monitoring was scheduled at 10 days post-plantation. Diameter and region of interest (ROI) (ImageJ, US) of organoids were measured.
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