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Hygromycin-B is a broad-spectrum antibiotic produced by the bacterium Streptomyces hygroscopicus. It inhibits protein synthesis in eukaryotic and prokaryotic cells by interfering with the translocation step in the ribosome.

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Lab products found in correlation

Hygromycin b, by Thermo Fisher Scientific (3 mentions) Hct116, by American Type Culture Collection (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Dmem high glucose media, by Thermo Fisher Scientific (2 mentions) Hek293 cell, by American Type Culture Collection (2 mentions) Mccoy s 5a modified media, by Thermo Fisher Scientific (2 mentions) Pen strep, by American Type Culture Collection (1 mentions) Rpmi 1640, by American Type Culture Collection (1 mentions) Rpmi 1640, by Promega (1 mentions) Mem nonessential amino acids, by American Type Culture Collection (1 mentions) Pen strep, by Thermo Fisher Scientific (1 mentions) Mem non essential amino acid, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Hek293t cell, by American Type Culture Collection (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Dithiothreitol (dtt), by Merck Group (1 mentions) Glutamine, by Thermo Fisher Scientific (1 mentions)

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Hygromycin (2 citations)

3 protocols using hygromycin b

1

Generating Stable IRE1α-Expressing INS-1 Cells

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INS-1s (rat insulinoma beta cells) were grown in RPMI, 10% FBS buffer (v/v), 1 mM sodium pyruvate, 10 mM HEPES, 2 mM glutamine and 50 μM β-mercaptoethanol on Poly-D-Lysine coated tissue culture flasks. IRE1α (murine) was cloned into a pcDNA5/FRT/TO. INS-1 FRT/TO cells were grown following the protocol above. INS-1 FRT/TO cells were transfected with 2 μg IRE1α-pcDNA5/FRT/TO and 2 μg pOG44 using Lipofectamine 2000 (Thermo Fisher). Cell media was exchanged the next day and cells were grown for another day before passaging. Selection was performed using 50 μg/mL Hygromycin-B (Thermo Fisher) over about two weeks until all untransfected cells have died and colonies have appeared in transfected cells. Stably integrated cells were maintained in RPMI, 10% FBS buffer (v/v), 1 mM sodium pyruvate, 10 mM HEPES, 2 mM glutamine and 50 μM β-mercaptoethanol, 25 μg/mL Hygromycin-B. HEK293 cells (ATCC, # CRL-1573) were grown in DMEM High Glucose media (Gibco) supplemented with 10% FBS. HCT-116 (ATCC, #CCL-247) cells were maintained in McCoy’s 5A modified media (Gibco) supplemented with 10% FBS. Non-diabetic human islets were obtained from Prodo Labs (Irvine, CA) and cultured in supplemented Prodo Islet Medium (PIM(S) from Prodo Labs). All cells lines were maintained at 37°C with 5% CO2.
Feldman H.C., Ghosh R., Auyeung V.C., Mueller J.L., Kim J.H., Potter Z.E., Vidadala V.N., Perera B.G., Olivier A., Backes B.J., Zikherman J., Papa F.R, & Maly D.J. (2021). ATP-Competitive Partial Antagonists of IRE1α’s RNase Segregate Outputs of the UPR. Nature chemical biology, 17(11), 1148-1156.
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2

Generating Stable IRE1α-Expressing INS-1 Cells

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INS-1s (rat insulinoma beta cells) were grown in RPMI, 10% FBS buffer (v/v), 1 mM sodium pyruvate, 10 mM HEPES, 2 mM glutamine and 50 μM β-mercaptoethanol on Poly-D-Lysine coated tissue culture flasks. IRE1α (murine) was cloned into a pcDNA5/FRT/TO. INS-1 FRT/TO cells were grown following the protocol above. INS-1 FRT/TO cells were transfected with 2 μg IRE1α-pcDNA5/FRT/TO and 2 μg pOG44 using Lipofectamine 2000 (Thermo Fisher). Cell media was exchanged the next day and cells were grown for another day before passaging. Selection was performed using 50 μg/mL Hygromycin-B (Thermo Fisher) over about two weeks until all untransfected cells have died and colonies have appeared in transfected cells. Stably integrated cells were maintained in RPMI, 10% FBS buffer (v/v), 1 mM sodium pyruvate, 10 mM HEPES, 2 mM glutamine and 50 μM β-mercaptoethanol, 25 μg/mL Hygromycin-B. HEK293 cells (ATCC, # CRL-1573) were grown in DMEM High Glucose media (Gibco) supplemented with 10% FBS. HCT-116 (ATCC, #CCL-247) cells were maintained in McCoy’s 5A modified media (Gibco) supplemented with 10% FBS. Non-diabetic human islets were obtained from Prodo Labs (Irvine, CA) and cultured in supplemented Prodo Islet Medium (PIM(S) from Prodo Labs). All cells lines were maintained at 37°C with 5% CO2.
Feldman H.C., Ghosh R., Auyeung V.C., Mueller J.L., Kim J.H., Potter Z.E., Vidadala V.N., Perera B.G., Olivier A., Backes B.J., Zikherman J., Papa F.R, & Maly D.J. (2021). ATP-Competitive Partial Antagonists of IRE1α’s RNase Segregate Outputs of the UPR. Nature chemical biology, 17(11), 1148-1156.
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3

Cell Culture Protocols for Immune and Pancreatic Cell Lines

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PBMC used in this study were obtained from healthy volunteers (Immunocore employees). The Jurkat NFAT luciferase T cell line was purchased from Promega and maintained in RPMI-1640 supplemented with 10% heat-inactivated FBS, 2 mM L-glutamine, 0.1 mM MEM nonessential amino acids, 1 mM sodium pyruvate, pen/strep (50 U/mL penicillin and 50 µg/mL strep), and 200 μg/mL hygromycin B. The Raji B cell lymphoma line was purchased from ATCC and maintained in R10 growth media (RPMI-1640 supplemented with 10% heat-inactivated FBS, 2 mM L-glutamine, and pen/strep). HEK-293T cells were purchased from ATCC and maintained in DMEM (4.5 g/L glucose) supplemented with 10% heat-inactivated FBS and 2 mM L-glutamine. The immortalized human pancreatic β cell lines, ECN90 and EndoC-βH2, were purchased from Univercell Biosolutions and maintained in Optiβ3 and Optiβ1 media, respectively, in tissue culture vessels precoated with β-coat (Univercell Biosolutions).
All standard media (RPMI-1640, DMEM), FBS, glutamine, MEM nonessential amino acids, sodium pyruvate, pen/strep, PHA, and PBS were purchased from Thermo Fisher Scientific. hygromycin B was purchased from Invitrogen. Dithiothreitol was from Sigma-Aldrich.
Curnock A.P., Bossi G., Kumaran J., Bawden L.J., Figueiredo R., Tawar R., Wiseman K., Henderson E., Hoong S.J., Gonzalez V., Ghadbane H., Knight D.E., O’Dwyer R., Overton D.X., Lucato C.M., Smith N.M., Reis C.R., Page K., Whaley L.M., McCully M.L., Hearty S., Mahon T.M, & Weber P. (2021). Cell-targeted PD-1 agonists that mimic PD-L1 are potent T cell inhibitors. JCI Insight, 6(20), e152468.
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