S2 and S2 R+ cell lines (DGRC) were cultured in M3 media (Sigma-Aldrich S8398) containing 10% insect medium supplement (Sigma-Aldrich I7267) at 25 °C. Media for S2 R+ cell lines contained additional yeast extract and bactopeptone. Transfection was performed with effectene transfection reagent (Qiagen). For P100 preparation, conditioned media were sequentially centrifuged at 240×g, 2000×g, 10,000×g and 100,000×g29 (link). After 100,000×g centrifugation, supernatant fraction (SNΔ) was concentrated with a filter column. The obtained P100 fraction contains EVs including exosomes, was washed with PBS, and then pelleted again by additional centrifugation at 100,000×g.
S8398
Manufactured by Merck Group
The S8398 is a general-purpose laboratory centrifuge capable of separating a variety of sample types. It features a compact design, programmable settings, and a robust construction to meet the needs of modern research and testing laboratories.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using s8398
1
Isolation and Purification of Extracellular Vesicles
2
Drosophila Cell Line Maintenance and Antibody Usage
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The ML-D17c3 cell line, stock 107, was obtained from the Drosophila Genomics Resource Center cell line repository (http://dgrc.cgb.indiana.edu/ ). Cells were grown on pretreated tissue culture dishes with extracellular matrix as previously described (Currie and Rogers 2011 (link)) in M3 media (S-8398, Sigma-Aldrich) containing 1 mg/mL of yeast extracts (Y-1000, Sigma-Aldrich) and 2.5 mg/mL of bactopeptone (211677, Difco), 10% FBS (SH30070.02, Hyclone), and 10 mg/mL of insulin (I0516-5ml Sigma-Aldrich). The HepG2 cell line was kindly provided by B. Graveley (Institute for Systems Genomics, UCONN Health Center, Farmington) and maintained in Dulbecco's modified Eagle's medium (DMEM) (SH30022.01, Hyclone) supplemented with 10% FBS and 1% penicillin/streptavidin (15140-163, Invitrogen).
Rabbit polyclonal anti-Histone H3 (ab1791) and mouse monoclonal anti-α-tubulin (clone DM1A) were obtained from Abcam and Sigma-Aldrich, respectively. Mouse monoclonal anti-KDel (ADI-SPA-827), mouse monoclonal anti-Shot (mABRod1), and mouse monoclonal anti-ninein (clone F-5) were purchased from Enzo Life Sciences, the Developmental Studies Hybridoma Bank and Santa Cruz, respectively.
Rabbit polyclonal anti-Histone H3 (ab1791) and mouse monoclonal anti-α-tubulin (clone DM1A) were obtained from Abcam and Sigma-Aldrich, respectively. Mouse monoclonal anti-KDel (ADI-SPA-827), mouse monoclonal anti-Shot (mABRod1), and mouse monoclonal anti-ninein (clone F-5) were purchased from Enzo Life Sciences, the Developmental Studies Hybridoma Bank and Santa Cruz, respectively.
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