Pen strep

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Pen/Strep is a sterile solution containing the antibiotics Penicillin and Streptomycin. It is commonly used in cell culture applications to prevent bacterial contamination.

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Lab products found in correlation

3 363 protocols using pen strep

Cell Line Culture Conditions Across Disciplines

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Cell lines are listed in Supplementary Table S1 along with their characteristics, genetic information, origin, and source. RPE1 cells were cultured in Dulbecco’s Modified Eagle Medium Nutrient Mixture F12 (Life technologies) supplemented with 10% FBS (Sigma) and 1% pen/strep (ThermoFisher). CPA and CPD cells were cultured in Keratinocyte-SFM supplemented with 2.5 µg prequalified human Epidermal Growth Factor 1–53 (EGF) (Life technologies), 25 mg Bovine Pituitary Extract (BPE) (Life technologies) and 0.5% penicillin/streptomycin (pen/strep) (ThermoFisher). FLO cells were cultured in Dulbecco’s Modified Eagle Medium (D MEM) (Sigma) supplemented with 10% Fetal Bovine Serum (FBS) (Sigma) and 1% pen/strep (ThermoFisher). JH-Eso-AD1 cells were cultured in Minimum Essential Medium (MEM) (Sigma) supplemented with 10% FBS and 1% pen/strep (ThermoFisher). OE33 and OE19 cells were cultured in Roswell Park MEMorial Institute (RPMI) Medium (Life technologies) supplemented with 10% FBS (Sigma) and 1% pen/strep (ThermoFisher). All cells were cultured in a humidified atmosphere with 5% CO₂ at 37 °C.

Scott S.J., Li X., Jammula S., Devonshire G., Lindon C., Fitzgerald R.C, & D’Avino P.P. (2021). Evidence that polyploidy in esophageal adenocarcinoma originates from mitotic slippage caused by defective chromosome attachments. Cell Death and Differentiation, 28(7), 2179-2193.

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Cell Culture Conditions for Various Cell Lines

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RPE1 cells were obtained from ATCC and cultured in Dulbecco's Modified Eagle Medium Nutrient Mixture F12 (Life technologies) supplemented with 10% FBS (Sigma) and 1% pen/strep (ThermoFisher). CPA and CPD cells were cultured in Keratinocyte-SFM supplemented with 2.5 µg prequalified human Epidermal Growth Factor 1-53 (EGF) (Life technologies), 25 mg Bovine Pituitary Extract (BPE) (Life technologies) and 0.5% penicillin/streptomycin (pen/strep) (ThermoFisher). FLO cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) (Sigma) supplemented with 10% Fetal Bovine Serum (FBS) (Sigma) and 1% pen/strep (ThermoFisher). JH-Eso-AD1 cells were cultured in Minimum Essential Medium (MEM) (Sigma) supplemented with 10% FBS and 1% pen/strep (ThermoFisher). OE33 and OE19 cells were cultured in Roswell Park MEMorial Institute (RPMI) Medium (Life technologies) supplemented with 10% FBS (Sigma) and 1% pen/strep (ThermoFisher).
All cells were cultured in a humidified atmosphere with 5% CO2 at 37°C.

Scott S.J., Li X., Jammula S., Devonshire G., Lindon C., Fitzgerald R.C., & D’Avino P.P. (2020). Evidence that polyploidy in esophageal adenocarcinoma originates from mitotic slippage caused by defective chromosome attachments.

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Cell Culture Protocols for Diverse Cell Lines

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Vero E6 cells (CRL-1586, American Type Culture Collection, ATCC, Manassas, VA) and Calu-3 cells (HTB-55, ATCC) were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Thermo Fisher Scientific) supplemented with 20% v/v of filtered fetal bovine serum (FBS, Thermo Fisher Scientific), 1% v/v penicillin/streptomycin (Pen/Strep, Thermo Fisher Scientific) and 1% v/v of GlutaMAX supplement (Thermo Fisher Scientific). Human epithelial colorectal adenocarcinoma cell line Caco-2 (HTB-37, ATCC) was cultured in Minimum Essential Medium (Thermo Fisher Scientific) supplemented with 20% FBS, 1% Pen/Strep, and 1% GlutaMAX supplement. Primary human umbilical vein endothelial cells (HUVEC), pooled from multiple donors, were supplied by Invitrogen (Thermo Fisher Scientific) cryopreserved at the end of the primary culture stage. These cells were cultured in adhesion in Medium 200 (M200, Thermo Fisher Scientific), supplemented with 1% v/v Large Vessel Endothelial Supplement (LVES 50x, Thermo Fisher Scientific) and 1% v/v Pen/Strep. Peripheral blood mononuclear cells were purified by Ficoll-Paque PREMIUM (Merck) gradient from healthy blood donors and grown in RPMI 1640 Medium (Thermo Fisher Scientific) supplemented with 10% FBS, 1% Pen/Strep, 1% GlutaMAX and 1% Hepes. For the experiments, cells were seeded in 6-well or 12-well plates and maintained at 37°C in a humidified 5% CO2 incubator.

Giannella A., Riccetti S., Sinigaglia A., Piubelli C., Razzaboni E., Di Battista P., Agostini M., Dal Molin E., Manganelli R., Gobbi F., Ceolotto G, & Barzon L. (2022). Circulating microRNA signatures associated with disease severity and outcome in COVID-19 patients. Frontiers in Immunology, 13, 968991.

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Cell Culture Protocols for Diverse Cell Lines

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Human embryonic kidney (HEK)-derived 293T, HEK 293S N-acetylglucosaminyltransferase I-negative (GnTI-), and HeLa-derived TZM-bl cells were maintained in complete Dulbecco’s Modified Eagle Medium (herein referred to as cDMEM) containing high-glucose Dulbecco’s Modified Eagle Medium (DMEM, Thermo Fisher, Waltham, MA), 1× Penicillin-Streptomycin (Pen Strep, Thermo Fisher) and 10% fetal bovine serum (FBS, Gemini Bio Products, West Sacramento, CA) at 37°C/5% CO₂. Canine thymus-derived CF2Th.CD4.CCR5 cells were maintained in DMEM containing 1× Pen Strep, L-glutamine, 500 μg/ml G418, 150 μg/ml Hygromycin, and 10% FBS at 37°C/5% CO₂. FreeStyle 293F and Expi293F cells (both Thermo Fisher) were maintained in Freestyle 293 Expression Medium and Expi293 Expression Medium, respectively, at 37°C/10% CO₂ with shaking at 120 RPM. Human Burkitt’s B cell lymphoma-derived Ramos cells were maintained in either DMEM containing 15% FBS and 1× Pen Strep or Roswell Park Memorial Institute 1640 medium (RPMI 1640, Thermo-Fisher) containing 10% FBS and 1× Pen Strep at 37°C/5% CO_2. In some cases, Ramos cells were engineered to stably express mature B cell receptors (BCR) IgM versions of VRC01, PGT145, and PGT128.

Cale E.M., Gorman J., Radakovich N.A., Crooks E.T., Osawa K., Tong T., Li J., Nagarajan R., Ozorowski G., Ambrozak D.R., Asokan M., Bailer R.T., Bennici A.K., Chen X., Doria-Rose N.A., Druz A., Feng Y., Joyce M.G., Louder M.K., O’Dell S., Oliver C., Pancera M., Connors M., Hope T.J., Kepler T.B., Wyatt R.T., Ward A.B., Georgiev I.S., Kwong P.D., Mascola J.R, & Binley J.M. (2017). Virus-like Particles Identify an HIV V1V2 Apex-Binding Neutralizing Antibody that Lacks a Protruding Loop. Immunity, 46(5), 777-791.e10.

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Cell Culture Conditions for Various Cell Lines

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SKW3 T cells (DSMZ) were cultured in RPMI-1640+GluMax (Thermo Fisher Scientific) complemented with 10% fetal bovine serum (FBS, Sigma-Aldrich), 10 mM HEPES and 50 U/mL Pen-Strep (Thermo Fisher Scientific) at 37 °C and 5% CO₂.
LentiX cells and 293T cells were cultured in DMEM (Thermo Fisher Scientific) supplemented with 10% FBS, 2 mM L-Glutamine, 10 mM HEPES and 50 U/mL Pen-Strep (Thermo Fisher Scientific) at 37 °C and 5% CO₂.
KG-1 cells (ATCC) were cultured in IMDM (Thermo Fisher Scientific) supplemented with 10% FBS and 50 U/mL Pen-Strep (Thermo Fisher Scientific) at 37 °C and 5% CO₂.
SF9 cells were cultured in SF900-III media (Thermo Fisher) supplemented with 10% FBS and 10 mg/mL gentamicin sulfate (Thermo Fisher) at 27 °C and atmospheric CO₂.
Hi5 cells were grown in insect cell culture medium (Expression Systems) supplemented with 10 mg/mL gentamicin sulfate (Thermo Fisher) at 27 °C and atmospheric CO₂.
Jurkat cell lines were cultured in RPMI 1640 supplemented with 10% FBS, 2 mM L-Glutamine, 50 U/mL Penicillin, 50 μg/mL Streptomycin, and 50 μM β-mercaptoethanol at 37 °C and 5% CO₂.
HEK293T cell line was cultured in DMEM supplemented with 10% FBS, 2 mM L-Glutamine, and 18 mM HEPES at 37 °C and 5% CO₂.

Zhao X., Kolawole E.M., Chan W., Feng Y., Yang X., Gee M., Jude K.M., Sibener L.V., Fordyce P.M., Germain R.N., Evavold B.D, & Garcia K.C. (2022). Tuning T cell receptor sensitivity through catch bond engineering. Science (New York, N.Y.), 376(6589), eabl5282.

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Culturing HEK293T and THP-1 Reporter Cells

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HEK293T wild type (ATCC CRL-11268) and THP-1 wild type (ATCC TIB-202) cells were purchased from Leibniz Institute DSMZ, Braunschweig, Germany. HEK293T cells were cultured in DMEM GlutaMAX™ (Thermo Fisher Scientific, Oberhausen, Germany) supplemented with 10% (v/v) heat-inactivated FBS (Thermo Fisher Scientific), 1% (v/v) pen-strep (10,000 U/mL, Thermo Fisher Scientific), and 10 µg/mL puromycin dihydrochlorid (Carl Roth, Karlsruhe, Germany) (reporter cells only) and incubated at 37°C in a humidified atmosphere of 5% CO₂ and 95% air. THP-1 cells were cultured in RPMI 1640 GlutaMAX™ medium (Thermo Fisher Scientific) supplemented with 10% (v/v) heat-inactivated FBS and 1% (v/v) pen-strep (10,000 U/mL), and incubated at 37 °C in a humidified atmosphere of 5% CO₂ and 95% air.
During screening, CEBPD::SEAP THP-1 reporter cells were cultured in RPMI 1640 medium without phenol red (Thermo Fisher Scientific), supplemented with 10% (v/v) heat-inactivated FBS, 1% (v/v) pen-strep (10,000 U/mL), and 2 mM glutamine (Thermo Fisher Scientific) and incubated at 37 °C in a humidified atmosphere of 5% CO₂ and 95% air.
The HEK293T and THP-1 reporter cells generated were tested for mycoplasma contamination using a mycoplasma detection kit (Lonza, Basel, Switzerland) after cell sorting.

Ullmann T., Luckhardt S., Wolf M., Parnham M.J, & Resch E. (2021). High-Throughput Screening for CEBPD-Modulating Compounds in THP-1-Derived Reporter Macrophages Identifies Anti-Inflammatory HDAC and BET Inhibitors. International Journal of Molecular Sciences, 22(6), 3022.

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Pkd1 Knockout MEF Proliferation

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Immortalized Pkd1^+/+ and Pkd1^−/− MEFs were plated at a density of 150,000 cells/well in DMEM (Gibco) supplemented with 0.5% FBS (Gibco) and 1% Pen/Strep (Gibco). After 16 h, medium was changed to control medium (DMEM, Gibco), supplemented with 10% FBS (Gibco), 1% Pen/Strep (Gibco), sodium pyruvate (1 mM; Gibco), sodium bicarbonate (44 mM; Sigma Aldrich), 25 mM glucose (Sigma Aldrich), and 4 mM glutamine (Gibco); glucose starvation medium (DMEM supplemented with 1 mM sodium pyruvate, 44 mM sodium bicarbonate, 10% FBS, 1% Pen/Strep and 4 mM glutamine); glutamine starvation medium (DMEM supplemented with 10% FBS, 1% Pen/Strep, 1 mM sodium pyruvate, 44 mM sodium bicarbonate, and 25 mM glucose) or glucose and glutamine starvation medium (DMEM supplemented with 10% FBS and 1% Pen/Strep, 1 mM sodium pyruvate, 44 mM sodium bicarbonate). After 24 and 48 h, cells were trypsinized and counted with an automated cell counter (Countess cell counter, Invitrogen). Pictures from each sample at both time points were taken with a white field microscope, using a ×10 objective.

Podrini C., Rowe I., Pagliarini R., Costa A.S., Chiaravalli M., Di Meo I., Kim H., Distefano G., Tiranti V., Qian F., di Bernardo D., Frezza C, & Boletta A. (2018). Dissection of metabolic reprogramming in polycystic kidney disease reveals coordinated rewiring of bioenergetic pathways. Communications Biology, 1, 194.

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Cell Culture Protocols for Cancer and Immortalized Cell Lines

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Rabbit cells (cell line RK13; ATCC CCL-37) and MDA-435 (cell line MDA-MB-435; ATCC HTB-129) and PANC-1 (32 (link)) human cancer cells were cultured in Dulbecco minimum essential medium (DMEM; Invitrogen, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, USA), 2 mM glutamine (Invitrogen, USA), and 100 μg/mL of penicillin-streptomycin (Pen/Strep; Invitrogen, USA). Hare cells (cell line HN-R; Friedrich Loeffler Institut, Germany), a spontaneously immortalized cell line derived from kidney (41 (link)), were cultured in 50% Iscove’s modified Dulbecco’s medium (IMDM) and 50% Ham’s F-12 medium supplemented with 10% FBS, 2 mM glutamine, and 100 μg of Pen/Strep per mL. All cultures were maintained at 37°C in a humidified 5% CO₂ incubator (Thermo Fisher, USA). The RL-5 rabbit CD4⁺ T cell line was cultured in RPMI 1640 (obtained from Gibco), also supplemented with 10% FBS, 2 mM glutamine, and 100 g/mL of Pen/Strep.

Águeda-Pinto A., Kraberger S., Everts A., Gutierrez-Jensen A., Glenn H.L., Dalton K.P., Podadera A., Parra F., Martinez-Haro M., Viñuelas J.A., Varsani A., McFadden G., Rahman M.M, & Esteves P.J. (2022). Identification of a Novel Myxoma Virus C7-Like Host Range Factor That Enabled a Species Leap from Rabbits to Hares. mBio, 13(2), e03461-21.

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Cell Line Characterization and Culture

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Human embryonic kidney cell line, 293 T and human CRC cell lines, CACO2, COLO320DM, SW480, SW620, COLO205, HT29, and RKO were purchased from the National Collection of Authenticated Cell Cultures (Shanghai, China). CACO2 and COLO320DM were KRAS/BRAF wild-type cell lines. SW480 and SW620 were KRAS mutant and BRAF wild-type cell lines. COLO205, HT29, and RKO were BRAF V600E-mutant cell lines. 293 T, HT29, and RKO cells were cultured in DMEM/high glucose medium (SH30022.01B, HyClone, Logan, UT, USA) supplemented with 10% FBS (16140071, Gibco, Paisley, UK) and 1% Pen/Strep (16140071, Gibco, Paisley, UK). CACO2 cells were cultured in DMEM/high glucose medium (HyClone) supplemented with 20% FBS (Gibco) and 1% Pen/Strep (Gibco). COLO320DM and COLO205 cells were cultured in RMPI 1640 (HyClone) supplemented with 10% FBS (Gibco) and 1% Pen/Strep (Gibco). 293 T, HT29, RKO, CACO2, COLO320DM, and COLO205 cells were cultured at 37 °C under 5% CO₂. SW480 and SW620 cells were cultured in Leibovitz’s L15 medium (11415064, Gibco, Paisley, UK) supplemented with 10% FBS (Gibco) and 1% Pen/Strep (Gibco) at 37 °C without CO₂. Short tandem repeat profiling and mycoplasma test were performed for all cell lines to confirm the cell identity and ensure the cells were free from contamination.

Liu M., Xu X., Peng K., Hou P., Yuan Y., Li S., Sun X., Shi Z., Zhang J., Dong Y., Liu Q., Ai L., Liang L., Gan L., Huang Q., Yu Y, & Liu T. (2022). Heparanase modulates the prognosis and development of BRAF V600E-mutant colorectal cancer by regulating AKT/p27Kip1/Cyclin E2 pathway. Oncogenesis, 11(1), 58.

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SARS-CoV-2 Isolation and Culture Protocols

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VeroE6 (ATCC CRL-1586) and Vero (ATCC CCL-81) cells were maintained in Dulbecco’s modified Eagle medium (DMEM; Sigma-Aldrich) supplemented with 10% FCS (Gibco, Gaithersburg, MD, USA), and 100 U/mL penicillin-streptomycin (Pen/Strep; Gibco) at 37°C and 5% CO₂. Rhileki cells were maintained in DMEM supplemented with 10% FCS, 1% nonessential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), and 100 U/mL Pen/Strep at 37°C and 5% CO₂.
SARS-CoV-2 isolation and culture was performed using Vero cells with DMEM containing 5% FCS, and 100 U/mL Pen/Strep, or using Rhileki cells with DMEM containing 0.5% bovine serum albumin (BSA; Gibco), 1% nonessential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), and 100 U/mL Pen/Strep. Isolation attempts were performed by incubating cells for up to 7 days (for Vero cells), or up to 15 days (for Rhileki cells) with filtrated (0.45 μm) combined nasopharyngeal/oropharyngeal swab samples from individual patients. For subsequent virus culture experiments of successfully isolated strains, inoculation was restricted to 1 h at 37°C and 5% CO₂, followed by a washing step of the cells with DMEM. Established Cambodian ancestral Wuhan SARS-CoV-2 isolates 1775 (GISAID: EPI_ISL_956384), 2018 (GISAID: EPI_ISL_956389), and 2310 (GISAID: EPI_ISL_956394) passaged up to six times in Vero cells were used for comparative cultivation experiments.

Auerswald H., Low D.H., Siegers J.Y., Ou T., Kol S., In S., Linster M., Su Y.C., Mendenhall I.H., Duong V., Smith G.J, & Karlsson E.A. (2022). A Look inside the Replication Dynamics of SARS-CoV-2 in Blyth’s Horseshoe Bat (Rhinolophus lepidus) Kidney Cells. Microbiology Spectrum, 10(3), e00449-22.

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