Fetal calf serum (fcs)

Manufactured by Roche

Sourced in Switzerland

The FCS is a specialized laboratory equipment used for the analysis and characterization of biological samples. It utilizes flow cytometry technology to rapidly detect and measure various properties of cells or particles suspended in a fluid stream. The FCS provides quantitative data on factors such as size, granularity, and fluorescence intensity, enabling researchers to investigate cellular characteristics and population dynamics.

Automatically generated - may contain errors

Lab products found in correlation

8 protocols using fetal calf serum (fcs)

CHO Cell Culture and DAMI Cell Differentiation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Chinese hamster ovary (CHO) cells expressing GPIbα, GPIbβ and GPIX on their surface (CHO GPIb-IX) or CHO cells expressing only GPIbβ and GPIX and not GPIbα (CHO β9) (both kind gifts from J.A. Lopez, Puget Sound Blood Center, Seattle, WA) were cultured in α Minimum Essential Medium (Life Technologies, Carlsbad, CA) supplemented with 10% Fetal Calf Serum, 1% Penicillin-Streptomycin and in the presence of G418 (Roche, Indianapolis, IN) and/or methotrexate (Sigma-Aldrich, St. Louis, MO) as previously described [18 (link)]. Human megakaryoblastic DAMI cells were obtained from ATCC (Manassas, VA) and grown in RPMI1640 medium supplemented with 10% Fetal Calf Serum, 1% Penicillin-Streptomycin, 1% MEM NEAA and 1% sodium-pyruvate (all from Life Technologies) at 37°C and 5% CO₂. For differentiation experiments, 1μM PMA (Merck, Darmstadt, Germany) was added to DAMI growth medium (hereafter referred to as differentiation medium).

Thijs T., Broos K., Soenen S.J., Vandenbulcke A., Vanhoorelbeke K., Deckmyn H, & Salles-Crawley I.I. (2015). Artificial MiRNA Knockdown of Platelet Glycoprotein lbα: A Tool for Platelet Gene Silencing. PLoS ONE, 10(7), e0132899.

+ Open protocol

+ Expand

Isolation and Culture of Podocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Kidneys dissected from 10-day-old triple transgenic Mof^fl/fl;Nphs2-Cre^T/+;tomato^fl/+>eGFP and littermate control mice were minced into 1-mm³ pieces and treated with 3 ml enzymatic digestion buffer (300 U/ml collagenase, 1 mg/ml pronase E, 50 U/ml DNase I) at 37 °C for 15 min. The digested kidneys were pressed through cell strainers with decreasing pore sizes (100, 70 and 40 μm) and washed extensively with Hank's balanced salt solution. Cells were collected by centrifugation, re-suspended in 0.5 ml of Hank's balanced salt solution supplemented with 0.1% bovine serum albumin plus 4'6-diamidino-2-phenylindole (1 μg/ml) before fluorescence-activated cell sorting. Collected podocytes were cultured in collagen IV-coated tissue culture flasks (Nunc) in RPMI medium supplemented with penicillin–streptomycin, fetal calf serum and insulin–transferrin–sodium selenite supplement (Roche Applied Science, Basel, Switzerland) at 37 °C in 95% O₂ and 5% CO₂.

Sheikh B.N., Bechtel-Walz W., Lucci J., Karpiuk O., Hild I., Hartleben B., Vornweg J., Helmstädter M., Sahyoun A.H., Bhardwaj V., Stehle T., Diehl S., Kretz O., Voss A.K., Thomas T., Manke T., Huber T.B, & Akhtar A. (2015). MOF maintains transcriptional programs regulating cellular stress response. Oncogene, 35(21), 2698-2710.

+ Open protocol

+ Expand

Immunofluorescence and Microvessel Density Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunofluorescence and image analysis were performed as described previously.22 (link) Briefly, in order to stain blood vessels, frozen sections of tissues were fixed in paraformaldehyde for 10 min, washed in Phosphate-buffered saline (PBS), and blocked with fetal calf serum (dilution, 1:20; Roche) for 20 min. Specimens were incubated overnight at 4°C with mouse anti-human CD31 monoclonal antibodies (dilution, 1:100; Dako), then washed in PBS and stained with goat anti-mouse secondary antibody Fitc (dilution, 1:50; Jackson) for 1 hour at room temperature. Then, DAPI nuclear dye was used to stain for 10 min. The doxorubicin auto-fluorescence and blood vessels were observed by a fluorescence microscope (Olympus, Tokyo, Japan) and captured by a camera (Q-IMAGING, British Columbia, Canada).
Microvessel density (MVD) was calculated by the method developed by Weidner et al.30 (link) First, the most intense vascularization of each tumor section was detected under a low magnification field (×40 magnification), then microvessels were counted at three high magnification fields (×100 magnification). The final MVD was the mean value of three fields. The calculation method of doxorubicin was similar to the method of MVD calculation. All these procedures were conducted by Image-Pro Plus 6.0 (Media Cybernetics, Rockville, MD, USA).

Xiong F., Cao Y., Guo X., Zhang H., Wang J., Xiong B., Liang B, & Zheng C. (2020). Improved ADM Penetration Distance and Therapeutic Efficiency in a Rabbit VX2 Liver Cancer Model by Relaxin Infusion Combined with Transcatheter Chemoembolization Through Hepatic Artery. Cancer Management and Research, 12, 3379-3388.

+ Open protocol

+ Expand

Isolation of Leukocytes from Murine and Human Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse uterus, spleen, liver, lung, PP, and lymph nodes were processed using enzymatic protocol. Minced tissues were incubated 2 × 15 min with HBSS (PAA), 10% FCS (Life Technologies), 5 mM EDTA (Sigma), 15 mM HEPES solution (Life Technologies) on a rotator at 37°C, then digested during 30 min with RPMI 1640 containing 2% FCS, 30 μg/ml DNase I (Roche), and 0.1 WU/ml Liberase DH (Roche). Digested tissues were filtered and smashed with a syringe plunger on a cell strainer to mechanically dissociate the remaining bits of tissues. Leukocytes were enriched on an 80%/40% Percoll (GE Healthcare Life Sciences) gradient. Human decidua and endometrium tissues were mechanically processed. Leukocytes were enriched by layering on Lymphoprep (Axis-Shield).

Doisne J.M., Balmas E., Boulenouar S., Gaynor L.M., Kieckbusch J., Gardner L., Hawkes D.A., Barbara C.F., Sharkey A.M., Brady H.J., Brosens J.J., Moffett A, & Colucci F. (2015). Composition, Development, and Function of Uterine Innate Lymphoid Cells. The Journal of Immunology Author Choice, 195(8), 3937-3945.

+ Open protocol

+ Expand

Isolation and Differentiation of Monocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Monocytes (Mo) were isolated in 6-well plates (Nuclon, Denmark) by adhesion of MNCs (3 × 10⁶ cells/mL) to the plastic in the presence of 5% human AB serum. Monocyte-derived macrophages (MDM) were generated by culturing adherent fraction of MNCs during 7 days in RPMI-1640 medium completed with 5% autoplasma, 2% fetal calf serum (FCS, Biolot, Russia), 2-mercaptoethanol (5 × 10⁻⁵ M, Serva, Germany), pyruvate Na (2 × 10⁻³ M, Sigma-Aldrich, USA), and 1% nonessential amino acid solution in the presence of GM-CSF (50 ng/mL, Sigma-Aldrich, USA). In 7 days macrophages were harvested using 0.25% trypsin/EDTA solution. Monocyte-derived dendritic cells (MDDC) were generated by culturing adherent fraction of MNCs during 4 days in RPMI-1640 medium with 5% FCS in the presence of GM-CSF (40 ng/mL) and IFN-α (1,000 U/mL, Roferon-A, Roche, Switzerland), followed by maturation over 24 hours in the presence of 10 μg/mL lipopolysaccharide (LPS E. coli 0111:B4, Sigma-Aldrich, USA).

Sakhno L.V., Shevela E.Y., Tikhonova M.A., Nikonov S.D., Ostanin A.A, & Chernykh E.R. (2015). Impairments of Antigen-Presenting Cells in Pulmonary Tuberculosis. Journal of Immunology Research, 2015, 793292.

+ Open protocol

+ Expand

Single Cell Isolation from Mouse Intestine

Check if the same lab product or an alternative is used in the 5 most similar protocols

To remove epithelial cells, the small intestine was incubated in HBSS (Gibco, Carlsbad, CA) containing 2 mM EDTA, 2% FCS (PAA Laboratories, Pasching, Austria), and 1 mM DTT (Sigma, St. Louis, MO). This treatment was repeated twice for 20 min at 37°C with gentle agitation. The tissue was washed once with HBSS to remove EDTA and then transferred to a digestion solution containing 10% FCS, 60 CDU/ml collagenase D (Roche), 60 kunitz units/ml DNase I (Roche, Basel, Switzerland), 0.28 CDU/ml Dispase (Roche), and 1 mM CaCl₂ in HBSS for 20 min at 37°C with gentle agitation. The contents were transferred to gentleMACS tubes (Miltenyi Biotech, Bergisch Gladbach, Germany) and disrupted on a gentleMACS dissociator (Miltenyi Biotech). After filtration, single cell suspensions were obtained. MLN and PP were transferred to 5 ml of the digestion solution, incubated at 37°C for 45 min with agitation and the digested organs were pipetted into single cell suspensions. Spleens were incubated with 1 ml of digestion solution for 30 min at 37°C and then pressed through a nylon mesh to obtain single cells. Erythrocytes were lysed with 2 ml of a hypotonic solution of NH₄Cl. All cell suspensions were washed once with HBSS and resuspended in sterile PBS. Cells were counted using a Sysmex KX-21N cell counter (Sysmex Corporation, Kobe, Japan).

Fernández-Santoscoy M., Wenzel U.A., Yrlid U., Cardell S., Bäckhed F, & Wick M.J. (2015). The Gut Microbiota Reduces Colonization of the Mesenteric Lymph Nodes and IL-12-Independent IFN-γ Production During Salmonella Infection. Frontiers in Cellular and Infection Microbiology, 5, 93.

+ Open protocol

+ Expand

Isolation of Immune Cells from Mouse Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse uterus, spleen, liver, lung, Peyer’s patches and lymph nodes were processed using enzymatic protocol. Minced tissues were incubated 2 × 15 min with HBSS (PAA), 10% FCS (Life Technologies), 5 mM EDTA (Sigma), 15 mM HEPES solution (Life Technologies) on a rotator at 37°C. Then digested during 30 min with RPMI containing 2% FCS, 30 μg/ml DNase I (Roche) and 0.1 WU/ml Liberase DH (Roche). Digested tissues were filtered and smashed with a syringe plunger on a cell strainer to mechanically dissociate the remaining bites of tissues. Leukocytes were enriched on a 80%/40% Percoll (GE Healthcare Life Sciences) gradient. Human decidua and endometrium tissues were mechanically processed. Leukocytes were enriched by layering on Lymphoprep (Axis-Shield).

+ Open protocol

+ Expand

Isolation of Lamina Propria Cells from Intestine

Check if the same lab product or an alternative is used in the 5 most similar protocols

Colon and small intestine were removed from mesenteric fat tissue, opened longitudinally and subsequently cleaned in cold phosphate-buffered saline (PBS) and Hank’s Balanced Salt Solution (HBSS) (Gibco) supplemented with fetal calf serum (FCS, 1%, Lonza) and Penicillin/Streptomycin (Gibco) (100 μg/mL) (= wash medium). Epithelial cells were removed by an 8-minute incubation on a magnetic stirrer in warm (37°C) wash medium supplemented with DTT (1 mM) and EDTA (1 mM). After a washing step, tissue was cut in pieces smaller than 3mm and digested by a 25- to 30-min incubation on a magnetic stirrer in warm (37°C) MEM α containing Penicillin/Streptomycin (100 μg/mL), β-mercaptoethanol (50 μM, Sigma-Aldrich®), FCS (5%), DNase (5 U/mL, Roche), Collagenase V (0.85 mg/mL, Sigma-Aldrich®), Collagenase D (1.25 mg/mL, Roche), Dispase (1 mg/mL, Gibco). After digestion, cells were filtered through a 70 μm cell strainer and lamina propria cells were purified using a Percoll (VWR) gradient (44 to 67%). Purified cells were washed and re-suspended in PBS supplemented with FCS (1%) and Penicillin/Streptomycin (100 μg/mL) and stained for Flow Cytometry.

Aguilera-Lizarraga J., Florens M.V., Viola M.F., Jain P., Decraecker L., Appeltans I., Cuende-Estevez M., Fabre N., Van Beek K., Perna E., Balemans D., Stakenborg N., Theofanous S., Bosmans G., Mondelaers S.U., Matteoli G., Martinez S.I., Lopez-Lopez C., Jaramillo-Polanco J., Talavera K., Alpizar Y.A., Feyerabend T.B., Rodewald H.R., Farre R., Redegeld F.A., Si J., Raes J., Breynaert C., Schrijvers R., Bosteels C., Lambrecht B.N., Boyd S.D., Hoh R.A., Cabooter D., Nelis M., Augustijns P., Hendrix S., Strid J., Bisschops R., Reed D.E., Vanner S.J., Denadai-Souza A., Wouters M.M, & Boeckxstaens G.E. (2021). Local immune response to food antigens drives meal-induced abdominal pain. Nature, 590(7844), 151-156.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!