S2 cells from Invitrogen were grown at 25°C in HyClone SFX-Insect Cell Culture Media with 1x penicillin and streptomycin (Sigma-Aldrich). About 20 million cells were harvested for each ChIP-seq or ChIP-nexus experiment. S2 sells were cross-linked with 1% formaldehyde for 10 minutes at room temperature. Formaldehyde was quenched by 0.125 M glycine for 5 minutes. Cells were washed with PBS, re-suspended in Orlando and Paro’s Buffer A (0.25% triton X-100, 10 mM EDTA, 0.5 mM EGTA, 10 mM Tris-HCl, pH 8.0) and rotated for 10 minutes at room temperature. Nuclei were spun down and re-suspended in RIPA buffer (10 mM Tris-HCl, pH 8.0; 140 mM NaCl; 0.1% SDS; 0.1% sodium deoxycholate; 0.5% sarkosyl; 1% Triton X-100). The chromatin was fragmented with a Bioruptor by two rounds of 15 min sonication at high power. Chromatin was cleared by centrifugation and the supernatant was used for ChIP.
Hyclone sfx insect cell culture media
Manufactured by Merck Group
HyClone SFX-Insect Cell Culture Media is a serum-free, protein-free, and chemically defined medium designed for the growth and maintenance of insect cell lines. The media provides a balanced formulation of essential nutrients, vitamins, and growth factors required for the optimal growth and proliferation of insect cells in culture.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Merck Group (2 mentions)
Streptomycin, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Hek293t cell, by American Type Culture Collection (2 mentions)
Nci h211, by American Type Culture Collection (1 mentions)
Nci h526, by American Type Culture Collection (1 mentions)
Nci h1963, by American Type Culture Collection (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Sclc cell lines, by American Type Culture Collection (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
4 protocols using hyclone sfx insect cell culture media
1
ChIP-seq and ChIP-nexus of Drosophila S2 Cells
2
Cell Culture Protocols for HEK293T, SCLC, and Drosophila S2 Cells
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HEK293T cells were obtained from American Type Culture Collection (ATCC) and then maintained with Dulbecco’s modified Eagle’s medium (Gibco, Gaithersburg, MD) containing 10% fetal bovine serum (FBS; Sigma-Aldrich). The SCLC cell lines NCI-H526, NCI-H211, NCI0H510, and NCI-H1963 were obtained from ATCC and were maintained with ATCC-formulated RPMI 1640 medium containing 10% FBS (Sigma-Aldrich). The Drosophila S2 cells were maintained in HyClone SFX-Insect Cell Culture Media containing 10% FBS (Sigma-Aldrich).
3
ChIP-seq and ChIP-nexus of Drosophila S2 Cells
4
Cell Culture Protocols for Various Cell Lines
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HEK293T cells were obtained from ATCC, and then maintained with DMEM (Gibco, Gaithersburg, MD) containing 10% FBS (Sigma). The SCLC cell lines were obtained from ATCC. NCI-H748, NCI-H1963, and NCI-H510 cells were maintained with ATCC-formulated RPMI-1640 medium containing 10% FBS (Sigma). The Drosophila S2 cells were maintained in HyClone™ SFX-Insect Cell Culture Media containing 10% FBS (Sigma).
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