Fetal calf serum (fcs)

Manufactured by Leibniz Institute DSMZ

Sourced in Germany

The FCS (Fluorescence-activated Cell Sorting) is a specialized laboratory instrument used for the analysis and sorting of cells or particles within a fluid sample. It utilizes laser technology and electronic detection to identify and separate specific cell types based on their physical and fluorescent characteristics.

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Lab products found in correlation

Du145, by Leibniz Institute DSMZ (2 mentions) Pa tu 8902, by Merck Group (1 mentions) Dulbecco s mem high glucose, by Merck Group (1 mentions) Mcf 7, by Merck Group (1 mentions) Mcf 7, by Leibniz Institute DSMZ (1 mentions) Pa tu 8902, by Leibniz Institute DSMZ (1 mentions) Nci h460, by Leibniz Institute DSMZ (1 mentions) Rpmi 1640 medium, by Leibniz Institute DSMZ (1 mentions) Ab2770, by Abcam (1 mentions) Ab3568, by Abcam (1 mentions) Ab33586, by Abcam (1 mentions) Fetal calf serum (fcs), by Harvard Bioscience (1 mentions) αmem glutamax, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) β mercaptoethanol, by Merck Group (1 mentions) Stemline 2 hematopoietic stem cell expansion medium, by Merck Group (1 mentions) Mccoy s 5a, by Leibniz Institute DSMZ (1 mentions) Mononuclear cell medium, by PromoCell (1 mentions) Magnetic activated cell sorting (macs), by Miltenyi Biotec (1 mentions) Tryptose phosphate broth, by Thermo Fisher Scientific (1 mentions)

5 protocols using fetal calf serum (fcs)

Culturing and Conditioning of Cancer Cell Lines

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Human breast carcinoma cell lines MCF-7 and HCC1143, pancreas adenocarcinoma cell line PA-TU-8902, prostate >carcinoma cell line DU145 and lung carcinoma cell line NCI-H460 were obtained from DSMZ (German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany).
In general RPMI 1640 medium, supplemented with 2 mM L-Glutamine and 1% Penicillin-Streptomycin and FCS (either 10% or 1% according to the assay) was used for monocyte/macrophage, NCI-H460, HCC1143 and DU145 cell culture. For MCF-7 cells we used Eagle´s MEM, supplemented with 10% FCS, 2 mM L-Glutamine, 1% Penicillin/Streptomycin and 1 mM Sodium Pyruvate and for PA-TU-8902 cells Dulbecco´s MEM High Glucose supplemented with 1 mM Sodium Pyruvate, 2 mM L-Glutamine, 1% Penicillin/Streptomycin and either 10% or 1% FCS was used (all reagents from Sigma, Buchs, Switzerland).
Cell lines were kept under standard culture conditions (37°C, 5% CO₂ and 95% humidity), tumour cell conditioned media were obtained by 24 h incubation of 90% confluent tumour cells in 10 ml of fresh culture medium in 25 cm² cell flasks. Cell free supernatants were carefully harvested, filtered through 0.5 μm filters and stored in aliquots at −80°C.

Estko M., Baumgartner S., Urech K., Kunz M., Regueiro U., Heusser P, & Weissenstein U. (2015). Tumour cell derived effects on monocyte/macrophage polarization and function and modulatory potential of Viscum album lipophilic extract in vitro. BMC Complementary and Alternative Medicine, 15, 130.

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Cytospin Immunostaining of Myeloid Cells

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Cytospin preparations of moDCs, LCs, MUTZ-3, THP-1 and U937 (~ 0.1 x 10 6 cells) were stained with monoclonal antibodies against CYP1A1 (ab3568), CYP1B1 (ab33586) and AhR (ab2770) (Abcam, Cambridge, UK) each for 1 h and protein expression was detected using a labeled streptavidin-biotin in Minimum Essential Medium alpha (MEM-α) + GlutaMAX™ (Gibco/Invitrogen, Darmstadt, Germany) supplemented with ribonucleosides and deoxyribonucleosides, 20% FCS (Biochrom, Berlin, Germany), 2 mM L-Glutamine (Life Technologies, Darmstadt, Germany), 50 µM β-mercaptoethanol (Sigma-Aldrich, Munich, Germany) and 10% of conditioned medium from the renal cell carcinoma cell line 5637 at 37°C in a 5% CO2 humidified incubator. Cells were subcultured at a split ratio of 1:2 twice per week. Cells were stimulated with benzo[a]anthracene on days 6 -7.
5637, a renal cell carcinoma cell line, was purchased from the German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany, ACC35) and was cultured in RPMI medium supplemented with 10% FCS. The cells were subcultured at a split ratio of 1:4 to 1:5 and passaged every 3-4 days.

Skazik-Voogt C., Kühler K., Ott H., Czaja K., Zwadlo-Klarwasser G., Merk H.F., Amann P.M, & Baron J.M. (2016). Myeloid human cell lines lack functional regulation of aryl hydrocarbon receptor-dependent phase I genes. ALTEX, 33(1).

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Culturing Leukemic Cell Lines and Primary Samples

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SEM, HL60, KCL22, K562 leukemic cell lines were cultured in RPMI1640 supplemented with 10% FCS and maintained at 37°C with 5% CO₂, except for SUP-B15 (BCRABL1) BCP-ALL cell line which was cultured in McCoy’s 5A supplemented with 20% of FCS (DSMZ, Braunschweig, Germany). Mononuclear cells (MNC) were isolated by Ficoll density gradient centrifugation using standard procedures and later cultured in Mononuclear Cell Medium (PromoCell, Heidelberg, Germany). CD34+ cells were later sorted from these MNC using MACS (Miltenyi Biotec, Bergisch Gladbach, Germany). Primary patient samples were obtained from newly diagnosed patients or from relapse after informed consent approval of the local ethics committee and were cultured either in Stemline® II Hematopoietic Stem Cell Expansion Medium (Sigma-Aldrich) or in Mononuclear Cell Medium (PromoCell)

Bhatia S., Krieger V., Groll M., Osko J.D., Reßing N., Ahlert H., Borkhardt A., Kurz T., Christianson D.W., Hauer J, & Hansen F.K. (2018). Discovery of the first-in-class dual histone deacetylase-proteasome inhibitor. Journal of medicinal chemistry, 61(22), 10299-10309.

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Culturing Melanoma and Cell Lines

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B16‐OVA melanoma (kindly provided by D.M. Brown22) was cultured in DMEM (Lonza, Switzerland) with 10% FCS (PAA Laboratories, Austria), 2% glutamine (Gibco, Waltham, MA), 100 U/ml penicillin (Gibco), 0.1 mg/ml streptomycin (Gibco) and 0.5 μg/ml geneticin (G418) sulfate (Santa Cruz Biotechnology, Dallas, TX). L929 cells (DSMZ GmbH, Germany) were cultured treated with either (i) PBS 10% FCS, 2% glutamine, 100 U/ml penicillin and 0.1 mg/ml streptomycin. BHK‐21 cells (ATCC, Manassas, VA) were cultured in GMEM (Gibco) with 10% FCS, 5% tryptose phosphate broth (Gibco), 100 U/ml penicillin and 0.1 mg/ml streptomycin.

Koske I., Rössler A., Pipperger L., Petersson M., Barnstorf I., Kimpel J., Tripp C.H., Stoitzner P., Bánki Z, & von Laer D. (2019). Oncolytic virotherapy enhances the efficacy of a cancer vaccine by modulating the tumor microenvironment. International Journal of Cancer, 145(7), 1958-1969.

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Cell Line Characterization and Maintenance Protocol

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All cell lines were obtained between 2018 and 2021, expanded, and stored as cryopreserved aliquots in liquid nitrogen. The human renal cell carcinoma SKRC52 cell line was kindly provided by Professor E Oosterwijk (Radbound University Nijmegen Medical Center, Nijmegen, The Netherlands) and maintained in RPMI medium (Invitrogen) supplemented with fetal calf serum (10%, FCS; Invitrogen) and antibiotic-antimycotic (1%, AA; Invitrogen). Chinese hamster ovary (CHO) cells, human fibrosarcoma HT1080, and murine colon carcinoma CT26 were purchased from the American Type Culture Collection. Cells were cultured according to the supplier’s protocol (for HT1080: Dulbecco’s Modified Eagle Medium+10% fetal bovine serum (FBS)+1% AA for CT26: RPMI+10% FBS+1% AA) and kept for no longer than 10 passages. Authentication of the cell lines was performed by the cell bank before shipment. SKRC52-hFAP cells and HT1080-hFAP were prepared as previously reported.14 (link) NK-92 cells were purchased from DSMZ (code ACC48) and cultured within MEM complete medium (75% α-MEM medium, 12.5% FCS, 12.5% horse serum, 2 mM L-glutamine, and IX antibiotic mix).

Nadal L., Peissert F., Elsayed A., Weiss T., Look T., Weller M., Piro G., Carbone C., Tortora G., Matasci M., Favalli N., Corbellari R., Di Nitto C., Prodi E., Libbra C., Galeazzi S., Carotenuto C., Halin C., Puca E., Neri D, & De Luca R. (2022). Generation and in vivo validation of an IL-12 fusion protein based on a novel anti-human FAP monoclonal antibody. Journal for Immunotherapy of Cancer, 10(9), e005282.

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