Media a

Manufactured by STEMCELL

Sourced in United States

Media A is a cell culture medium designed for the growth and maintenance of various cell types. It provides the necessary nutrients and growth factors to support cell proliferation and viability. The composition of Media A is optimized to support the specific requirements of the targeted cell lines.

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Lab products found in correlation

Media e, by STEMCELL (1 mentions) Serum free media, by Thermo Fisher Scientific (1 mentions) Protein g column, by GE Healthcare (1 mentions) Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)

2 protocols using media a

Hybridoma Generation from PBMC B Cells

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Eight days following plating of PBMCs, wells identified to contain positive B cells by ELISA were selected for electrofusion to generate hybridomas as previously described (24 , 25 ). Hybridomas were plated in 384-well plates for HAT selection, and grown for 14 days at 37°C, 5% CO₂. Following screening by ELISA, hybridomas were single-cell sorted using a MoFlo Astrios cell sorter using live/dead staining by propidium iodide. The sorted hybridomas were cultured in 25% Media E (StemCell) + 75% Media A (StemCell) for two weeks, then subjected to another round of screening by ELISA. Hybridomas with the highest signal were grown in 250 mL serum-free media (Gibco) for approximately one month. Secreted mAbs were purified using a Protein G column (GE Healthcare) and concentrated for use in downstream assays.

Nagashima K., Dzimianski J.V., Han J., Abbadi N., Gingerich A.D., Royer F., O’Rourke S., Sautto G.A., Ross T.M., Ward A.B., DuBois R.M, & Mousa J.J. (2022). The pre-existing human antibody repertoire to computationally optimized influenza H1 hemagglutinin vaccines. Journal of immunology (Baltimore, Md. : 1950), 209(1), 5-15.

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Culturing Insect and Mammalian Cell Lines

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ExpiSf9 cells (Invitrogen, Waltham, MA, USA) were maintained in suspension culture in ExpiSf^TM CD Medium (Thermo Fisher Scientific, Carlsbad, CA, USA) at 28 °C in a non-humidified, non-CO₂ incubator JSSI-200C (JS Research Inc., Gongju, Korea) with shaking at 120 rpm. ExpiSf9 cells were subcultured at a density of 0.5 × 10⁶ cells/mL every 4 days. The Sp2/0-Ag14 cell line (ATCC, Manassas, VA, USA) was maintained in Media A (STEMCELL Technologies, Cambridge, MA, USA) and Dulbecco’s Modified Eagle Medium supplemented with 10% fetal bovine serum (FBS, Hyclone, Logan, UT, USA) and 1% antibiotic–antimycotic (Gibco, Rockville, MD, USA) at 37 °C in a 5% CO₂ incubator.

Jang J.H., Kim S., Kim S.G., Lee J., Lee D.G., Jang J., Jeong Y.S., Song D.H., Min J.K., Park J.G., Lee M.S., Han B.S., Son J.S., Lee J, & Lee N.K. (2022). A Sensitive Immunodetection Assay Using Antibodies Specific to Staphylococcal Enterotoxin B Produced by Baculovirus Expression. Biosensors, 12(10), 787.

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