Hi fbs

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom, Canada, Australia

The HI-FBS is a laboratory equipment product offered by Thermo Fisher Scientific. It is designed to perform high-intensity filtration and separation tasks. The core function of the HI-FBS is to efficiently filter and separate materials in a laboratory setting, though its intended use is not specified.

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Lab products found in correlation

302 protocols using hi fbs

Maintenance of Cell Lines for Viral Transduction

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Ba/F3 cells were purchased from DSMZ and maintained in 90% RPMI (Gibco), 10% HI-FBS (Gibco), 1% penicillin/streptomycin (Gibco), and 10ng/ml IL-3 (Fisher), and incubated at 37°C with 5% CO2. Ba/F3 cells were passaged at or below 1.0E6 cells/ml in order to avoid acquired IL-3 resistance, and regularly checked for IL-3 addiction by performing 3x PBS (Gibco) washes and outgrowth in the absence of IL-3 to confirm cell death in the parental, empty cell line.
Plat-E cells stably expressing retroviral envelope and packaging plasmids were originally gifted by Dr. Wendell Lim. Plat-E cells were maintained in 90% DMEM+HEPES (Gibco), 10% HI-FBS (Gibco), 1% penicillin/streptomycin (Gibco), 10ug/ml blasticidin, 1ug/ml puromycin, and incubated at 37°C with 5% CO2. Plat-E cells were maintained under blasticidin and puromycin antibiotic pressure unless being transfected.
HEK293 cells were maintained in DMEM (Gibco) supplemented with 10% FBS (Gibco) and 1% penicillin/streptomycin (Gibco) at 37°C in 5% CO2.
Human MET knockout HeLa cells were purchased from Abcam and maintained in 90% DMEM+HEPES (Gibco), 10% HI-FBS (Gibco), 1% penicillin/streptomycin (Gibco), and incubated at 37°C with 5% CO2

Estevam G.O., Linossi E.M., Macdonald C.B., Espinoza C.A., Michaud J.M., Coyote-Maestas W., Collisson E.A., Jura N, & Fraser J.S. (2023). Conserved regulatory motifs in the juxtamembrane domain and kinase N-lobe revealed through deep mutational scanning of the MET receptor tyrosine kinase domain. bioRxiv.

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CRISPRi Validation in K562 Cells

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K562 cells with a doxycycline-inducible CRISPRi were a gift of the Lander lab and identical to those used in previous studies^{17 (link)}. Cells were grown in RPMI 1640 GluteMAX (Gibco) with 10% heat inactivated FBS (HI-FBS, Gibco). Cells were induced with CRISPRi for 24 h with a final concentration of 1 μg/ml doxycycline (VWR). In tests of guides for FADS, GATA1, and MYC, doxed and undoxed distributions were highly correlated (Spearman ρ = 0.91, 0.80, and 0.86, respectively). Non lenti-library infected cells were periodically sorted on BFP signal when BFP/CRISPRi expression dropped below 80%. Jurkat and GM12878 cells were grown in RPMI 1640 GluteMAX with 15% HI-FBS, 293T cells were grown in DMEM (Gibco) with 10% HI-FBS, SK-N-SH cells were grown in EMEM (ATCC) with 10% HI-FBS, and TF1 cells were grown in RPMI 1640 GluteMAX (Gibco) with 10% heat-inactivated FBS, 2 mM L-Glutamine and 2 ng/ml recombinant human Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) (Peprotech). All cells were grown at 37 °C and 5% CO₂. Cells were not synchronized prior to performing HCR-FlowFISH.

Reilly S.K., Gosai S.J., Gutierrez A., Mackay-Smith A., Ulirsch J.C., Kanai M., Mouri K., Berenzy D., Kales S., Butler G.M., Gladden-Young A., Bhuiyan R.M., Stitzel M.L., Finucane H.K., Sabeti P.C, & Tewhey R. (2021). Direct characterization of cis-regulatory elements and functional dissection of complex genetic associations using HCR-FlowFISH. Nature genetics, 53(8), 1166-1176.

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Cell Line Culturing Protocols

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SW780 cells were purchased from ATCC (#CRL-2169) and RT112 cells were purchased from ECACC (#85061106). RT4 cells were kindly provided by Dr. Darryl Martin’s lab (Yale University, New Haven, CT, USA). UM-UC-14 cells were a generous gift from Dr. Sharada Mokkapati’s lab (MD Anderson Cancer Center, Huston, TX, USA). TRT-HU-1 cells were kindly provided by Dr. Rosalyn M. Adam (Harvard Medical School, Boston, MA, USA). SW780 cells were cultured in DMEM media (Gibco) supplied with 10% heat-inactivated FBS (HIFBS, Gibco) and 1% antibiotic-antimycotic (A/A, Gibco). RT112 cells were maintained in DMEM media supplied with 10% HIFBS, 1% A/A, 1% non-essential amino acid (NEAA, Gibco), and 2 mM L-glutamine (Gibco). RT4 cells were cultured in McCoy’s 5A (ATCC) media supplied with 10% heat-inactivated FBS (HIFBS) and 1% antibiotic-antimycotic (A/A). UM-UC-14 cells were maintained in DMEM media supplied with 10% HIFBS, 1% A/A, 1% NEAA, 2 mM L-glutamine, and 1 mM sodium pyruvate (Gibco). TRT-HU-1 cells were cultured in DMEM media supplied with 15% non-heat inactivated FBS (Gibco), 1% A/A, 1% NEAA, 110 mg/L sodium pyruvate (Gibco), 1.15 mM 1-thioglycerol (Sigma), and 2 mM L-glutamine (Gibco). All cells were cultured at 37 °C in a humidified atmosphere with 5% CO₂ and tested to be mycoplasma negative.

Wang Z., Muthusamy V., Petrylak D.P, & Anderson K.S. (2023). Tackling FGFR3-driven bladder cancer with a promising synergistic FGFR/HDAC targeted therapy. NPJ Precision Oncology, 7, 70.

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Cultivation of Leishmania infantum Promastigotes

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We used promastigotes of L.
infantum lines: (i) JPC-M5 (MCAN/ES/98/LLM-877) (LJPC)
as a genomic reference line and (ii) LLM2070, LLM2165, LLM2255, and
LLM2221 lines (L2070, L2165, L2255, and L2221), isolated from TF HIV
patients with VL and unsuccessfully treated with liposomal AmB (from
the WHO Collaborating Center for Leishmaniasis, Instituto de Salud
Carlos III; Dr. F. Javier Moreno). All these L. infantum lines were grown at 28 °C in the RPMI 1640-modified medium
(Invitrogen) supplemented with 10% hiFBS (Invitrogen), as described.^{45 (link)} Human myelomonocytic cells THP-1 were grown
at 37 °C and 5% CO₂ in the RPMI-1640 medium supplemented
with 10% hiFBS, 2 mM glutamate, 100 U/mL penicillin, and 100 mg/mL
streptomycin, as described.^{46 (link)}

Perea-Martínez A., García-Hernández R., Manzano J.I, & Gamarro F. (2022). Transcriptomic Analysis in Human Macrophages Infected with Therapeutic Failure Clinical Isolates of Leishmania infantum. ACS Infectious Diseases, 8(4), 800-810.

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Cultivation of Trypanosoma Parasites and Enterococcus Bacterium

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‘Single marker’ (S16) T. brucei (Lister 427, antigenic type MiTat 1.2, clone 221a) BSF and T. brucei 449 PCF were cultured as previously described (Wirtz et al., 1999 (link); Cabello-Donayre et al., 2016 (link)). T. brucei rhodesiense (EATRO3 ETat1.2 TREU164) and T. brucei gambiense (ELIANE strain) BSF were grown at 37 °C, 5% CO₂ in HMI-9 medium supplemented with 20% heat-inactivated fetal bovine serum (hiFBS, Invitrogen). To generate clathrin depleted cell lines, the plasmid p2T7TiCLH was transfected into the T. b. brucei S16 cell line (Allen et al., 2003 (link)). The human MRC-5 cell line (fibroblasts derived from lung tissue) was cultured in DMEM (Invitrogen) medium plus 10% hiFBS. The producer AS-48 Enterococcus faecalis UGRA-10 strain (Cebrián et al., 2012 (link)) was growth in Brain Hearth Infusion (BHI) or Brain Hearth Agar (BHA) (Merck) media at 37 °C without aeration. For AS-48 production, UGRA-10 strain was cultured at 28 °C in Esprion 300 (E−300, DMV Int., Veghel, Netherland) plus 1% glucose (E−300-G) following the conditions established by Ananou et al. (2008) (link).

Martínez-García M., Bart J.M., Campos-Salinas J., Valdivia E., Martínez-Bueno M., González-Rey E., Navarro M., Maqueda M., Cebrián R, & Pérez-Victoria J.M. (2018). Autophagic-related cell death of Trypanosoma brucei induced by bacteriocin AS-48. International Journal for Parasitology: Drugs and Drug Resistance, 8(2), 203-212.

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Canine Neoplastic Cell Line Protocol

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Three canine neoplastic primary cell lines were used, representing round, mesenchymal and epithelial tumor types: mastocytoma C2 (University of California, San Fransisco, USA), mammary gland carcinoma CMT-12 (Auburn University, Alabama, USA), and osteosarcoma D17 (#CCL-183; ATCC, Manassas, VA, USA). The cell lines were grown on tissue culture-treated plates (Laboratory Product Sales [LPS], Rochester, NY, USA) with appropriate medium containing 10 % heat inactivated fetal bovine serum (HI-FBS; Invitrogen, Carlsbad, CA, USA) and 1 % antibiotic-antimycotic (Invitrogen). Cell lines were grown at 37 °C and 5 % CO₂ for all experiments and passage of cells, unless otherwise noted. Canine primary dermal fibroblasts (Applied Biological Materials [ABM], Richmond, BC, Canada) were used to investigate effects on normal cells and were propagated and maintained on PriCoat T25 flasks (ABM) in Prigrow II medium (ABM) containing 10 HI-FBS (Invitrogen) and 1 % penicillin/streptomycin (Invitrogen).

Levine C.B., Bayle J., Biourge V, & Wakshlag J.J. (2016). Effects and synergy of feed ingredients on canine neoplastic cell proliferation. BMC Veterinary Research, 12, 159.

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Recombinant Rotavirus Protein Expression

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MA104 cells (ATCC) were cultured in M199 media (Invitrogen) supplemented with 25 mM HEPES and 10% HI-FBS (Invitrogen). BSC-1 cells (ATCC) were cultured in DMEM (Invitrogen) supplemented with 10% HI-FBS (Invitrogen). For VP4 and VP7 expression, full-length genomic sequences from rhesus rotavirus (G3 serotype, NCBI:txid444185) were amplified by PCR and cloned into pFastbac (Thermo Fisher Scientific) expression vector. Mutations were introduced by quick-change mutagenesis in DH10α cells (Thermo Fisher Scientific). Purified plasmid constructs were transfected into DH10-Bac cells (Thermo Fisher Scientific). Purified bacmids were transfected into SF9 cells (ATCC) grown in SF900 II SFM media supplemented with 1% pen-strep.

Herrmann T., Torres R., Salgado E.N., Berciu C., Stoddard D., Nicastro D., Jenni S, & Harrison S.C. (2021). Functional refolding of the penetration protein on a non-enveloped virus. Nature, 590(7847), 666-670.

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Transient Transfection of Cell Lines

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Cells were cultured in six-well plates at 37°C in 5% CO₂. RKO (CRL-2577; American Type Culture Collection [ATCC]) cells were grown in DMEM (Life Technologies) supplemented with 10% heat-inactivated fetal bovine serum (hi-FBS; Invitrogen). MCF7 (HTB-22; ATCC) cells were grown in DMEM:F-12 (Life Technologies) supplemented with 10% hi-FBS (Invitrogen). Reverse transfections of siRNA complexes was performed in cells seeded at 50% confluence with Lipofectamine RNAiMAX (ThermoFisher Scientific) in Opti-MEM (Life Technologies), according to the manufacturer’s instructions. Twenty-four hours after seeding, transfections of plasmid DNA was performed with Lipofectamine 3000 and P3000 reagent (ThermoFisher Scientific), according to manufacturer’s instructions. When required, the final amount of DNA used for transfection was kept constant by the addition of control vector DNA. All cells were harvested 48 h after transient DNA plasmid transfection for subsequent assays.

Hall E.T., Hoesing E., Sinkovics E, & Verheyen E.M. (2019). Actomyosin contractility modulates Wnt signaling through adherens junction stability. Molecular Biology of the Cell, 30(3), 411-426.

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Recombinant Rotavirus Protein Expression

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Culturing Murine and Human Cell Lines

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Murine brain microglial cells (BV-2) passage 18 (P:18) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Life technologies, Grand Island, NY) supplemented with 5% heat in activated fetal bovine serum (Hi-FBS, Invitrogen Corp., Carlsbad, CA) and 1% antibiotics (penicillin/streptomycin) (Invitrogen Corp., Carlsbad, CA). Human retinal pigment epithelial cells (ARPE-19) passage 21 (P: 21) were cultured in DMEM/F12(1:1) (Life technologies, Grand island, NY) supplemented with 10% Hi-FBS and 1% antibiotics. The above mentioned cell cultures were in a humidified incubator at 37 °C with 5% CO₂.

Kambhampati S.P., Mishra M.K., Mastorakos P., Oh Y., Lutty G.A, & Kannan R.M. (2015). Intracellular delivery of dendrimer triamcinolone acetonide conjugates into microglial and human retinal pigment epithelial cells. European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V, 95(Pt B), 239-249.

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