Pen strep

Manufactured by Merck Group

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Pen/Strep is a sterile solution containing a combination of penicillin and streptomycin antibiotics. It is commonly used in cell culture applications to prevent bacterial contamination.

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Close spelling variants of this product

Pen/Strep/Amphotericin (2 citations) Streptomycin (pen/strep, (4 citations) Pen-Strep solution (19 citations) Glutamine-Pen-Strep (2 citations) Penicillin-Streptomycin (Pen-Strep) (51 citations) Pen-Strep-Glut (2 citations) Pen/strep/neo antibiotics (1 citations) Pen/strep/myc (2 citations) Pen/Strep antibiotics (14 citations) Penicillin-streptomycin (pen/strep) solution (10 citations)

599 protocols using pen strep

Cultivation of Diverse Breast Cancer Cell Lines

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The MDA-MB-231 cells were cultured in DMEM/F12 (Sigma Aldrich, St. Louis, MO, USA; D8437) with 10% FBS (Sigma Aldrich, St. Louis, MO, USA; F2442) and 1% Pen/Strep (Sigma Aldrich, St. Louis, MO, USA; P4333). BT549 cells were cultured in RPMI-1640 (Sigma Aldrich, St. Louis, MO, USA; R8758) with 10% FBS and 1% Pen/Strep. MDA-MB-157 cells were cultured in DMEM (Sigma Aldrich, St. Louis, MO, USA; D6429) with 10% FBS and 1% Pen/Strep. The HMT-3522 S1 cells were cultured in DMEM/F12 with 250 ng/mL insulin, 10 μg/mL transferrin, 2.6 ng/mL sodium selenite, 10⁻¹⁰ M β-estradiol, 1.4 μM hydrocortisone, 5 μg/mL prolactin, and 10 ng/mL EGF. MCF-10A cells were cultured in DMEM/F12 with 5% horse serum, 20 ng/mL EGF, 0.5 mg/mL hydrocortisone, 100 ng/mL cholera toxin, 10 μg/mL insulin, and 1% Pen/Strep. THP-1 cells were cultured in RPMI-1640 with 10% FBS, 1% Pen/Strep, and 0.05 mM 2-mercaptoethanol. HEK293 FT cells (A kind gift from Dr. Mina J Bissell, Lawrence Berkeley National Laboratory) were cultured in DMEM with 10% FBS, 0.1 mM Non-Essential Amino Acids (Sigma Aldrich, St. Louis, MO, USA; M7145), 6 mM L-glutamine (VWR, Atlanta, GA, USA; 20J1956675), 1 mM Sodium Pyruvate (Sigma Aldrich, St. Louis, MO, USA; S8636), and 1% Pen/Strep. The cells were grown in a humidified incubator at 37℃ with 5% CO₂. All the cells were tested for mycoplasma contamination every two months.

Mao W., Xiong G., Wu Y., Wang C., St. Clair D., Li J.D, & Xu R. (2021). RORα Suppresses Cancer-Associated Inflammation by Repressing Respiratory Complex I-Dependent ROS Generation. International Journal of Molecular Sciences, 22(19), 10665.

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Lung Cancer Cell Culture Protocols

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A549 (NCI), SK-MES-1 and Calu-1 (both ATCC) were cultured in RMPI-1640, supplemented with 10% FBS, 1%Pen/Strep and 1%l-Glutamine (all from Sigma). NIC-H23 cells were cultured in RMPI-1640, supplemented with 10% FBS, 1%Pen/Strep, 1%l-Glutamine and 1 mM sodium pyruvate (Sigma). LL/2, Ladi 3.1 and Ladi 2.1 cells were cultured in DMEM (Sigma) supplemented with 10% FBS, 1%Pen/Strep and 1%l-Glutamine. Human cells were STR-profiled, used between passages 3 and 15, examined for mycoplasma and maintained in Plasmocin (Invivogen) to prevent contamination.

Siddiqui M.A., Gollavilli P.N., Ramesh V., Parma B., Schwab A., Vazakidou M.E., Natesan R., Saatci O., Rapa I., Bironzo P., Schuhwerk H., Asangani I.A., Sahin O., Volante M, & Ceppi P. (2020). Thymidylate synthase drives the phenotypes of epithelial-to-mesenchymal transition in non-small cell lung cancer. British Journal of Cancer, 124(1), 281-289.

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In Vitro Calcification Assay

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For in vitro calcification, hVICs and pVICs were seeded on 12-well plates and cultured with DMEM (Thermo Fisher Scientific, #21885108), 5% FBS (Gibco, #A3160802), and 1% penicillin/streptomycin (pen/strep) (Merck, #516106), osteogenic medium (OM), or pro-calcifying medium (PCM). OM consisted of DMEM with 5% FBS, 1% pen/strep, 10 nmol/L β-glycerophosphate (Sigma-Aldrich, #G9422), 10 mmol/L dexamethasone (Sigma-Aldrich, #D4902), and 50 µg/ml L-ascorbic acid (Carl Roth, #3525.2). PCM consisted of DMEM with 5% FBS, 1% pen/strep, 2 mmol/L sodium dihydrogen phosphate (NaH₂PO₄; Merck, #71507), and 50 µg/ml L-ascorbic acid (Carl Roth, #3525.2). Cells were incubated for 7 days to analyze gene expression by RT-qPCR and for 21 days for calcium deposition staining with 2% alizarin red (Sigma-Aldrich, #A5533). Cells were fixed with 4% formaldehyde for 15 min and washed twice with distilled water. Calcium nodules were stained with alizarin red for 15 min at room temperature, followed by two additionally washed. Staining was quantified by incubating cells with 3.58% hexadecylpyridinium chloride monohydrate (CPC) dissolved in ddH₂O for 1 h. The absorbance of 100 µl supernatant was measured at 550 nm using an Infinite® M Plex microplate reader (Tecan).

Nehl D., Goody P.R., Maus K., Pfeifer A., Aikawa E., Bakthiary F., Zimmer S., Nickenig G., Jansen F, & Hosen M.R. (2023). Human and porcine aortic valve endothelial and interstitial cell isolation and characterization. Frontiers in Cardiovascular Medicine, 10, 1151028.

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Stable Keratinocyte Cell Lines and T-Cell Culture

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shC and shFLG HaCaT keratinocytes established by us previously as a stable line by small harpin interference with the use of a lentiviral system (25 (link), 30 (link)) were grown in Dulbecco’s Modified Eagle’s Medium (DMEM-high glucose, Sigma-Aldrich) with 10% FBS (Sigma-Aldrich), 2 mM L-Glutamine (Sigma-Aldrich) and 1% Pen/Strep (Sigma-Aldrich) with selection carried out by puromycin at concentration of 20 µg/ml. K562-CD1a cells (a kind gift from Prof. Branch Moody) were cultured in RPMI-1640 (Sigma-Aldrich) with the addition of 200 µg/ml G418 (Thermo Fisher Scientific), 1% Pen/Strep (Sigma-Aldrich), and 10% heat-inactivated FBS (Sigma-Aldrich) and cultured at 37°C and 5% CO₂. For EV isolation media containing sEV-depleted FBS, treatments were carried out when the cells reached 80%–90% confluence (with the conditioned media being collected at 100% cell confluence; cell count in a region of 25 × 10⁶ cells per T75 flask). T cell medium was prepared by supplementing RPMI-1640 (Sigma-Aldrich) with 5% human male heat-inactivated AB serum (Sigma-Aldrich), 1% Pen/Strep (Sigma-Aldrich), 10 mM HEPES (Sigma-Aldrich), 2 mM L-Glutamine (Sigma-Aldrich), 1% non-essential amino acids (Biowest), 50 µM 2-mercaptoethanol (Sigma-Aldrich), and 10 ng/ml IL-2 (PeproTech).

Kobiela A., Hewelt-Belka W., Frąckowiak J.E., Kordulewska N., Hovhannisyan L., Bogucka A., Etherington R., Piróg A., Dapic I., Gabrielsson S., Brown S.J., Ogg G.S, & Gutowska-Owsiak D. (2024). Keratinocyte-derived small extracellular vesicles supply antigens for CD1a-resticted T cells and promote their type 2 bias in the context of filaggrin insufficiency. Frontiers in Immunology, 15, 1369238.

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Cell Culture Conditions for Various Cell Lines

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The human embryonic kidney epithelial cell line HEK293T (ATCC, CRL-3216), and human lung epithelial cell line A549 (ATCC, CCL-185) were cultured in Dulbecco’s modified Eagle’s medium (DMEM; high glucose, Sigma-Aldrich) supplemented with 10% (v/v) fetal bovine serum (FBS) (Sigma-Aldrich) and 1% (v/v) PenStrep (Sigma-Aldrich) and maintained in a 5% CO₂ incubator at 37°C. THP1 (ATCC, CRL-TIB-202) cells were cultured in Roswell Park Memorial Institute (RPMI)–1640 medium (R8758, Sigma-Aldrich) supplemented with 10% (v/v) FBS (Sigma-Aldrich) and 1% (v/v) PenStrep (Sigma-Aldrich) and maintained in a 5% CO₂ incubator at 37°C. Calu3 cells (ATCC, HTB-55) were cultured in Eagle’s minimum essential medium (302003, ATCC) supplemented with 10% (v/v) FBS (Sigma-Aldrich) and 1% (v/v) PenStrep (Sigma-Aldrich) and maintained in a 5% CO₂ incubator at 37°C. All cell lines were authenticated using STR profiling and confirmed to be free from mycoplasma contamination by testing with a mycoplasma detection kit (Sigma-Aldrich).

Khan S., Shafiei M.S., Longoria C., Schoggins J.W., Savani R.C, & Zaki H. (2021). SARS-CoV-2 spike protein induces inflammation via TLR2-dependent activation of the NF-κB pathway. eLife, 10, e68563.

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Adipocyte Differentiation with Heparin

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Cells were seeded on a 24-well plate at a density of 30,000 cells/cm². Cells were allowed to grow to confluence for 48 h in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS containing FFA and 1% Pen-Strep (Sigma). At this point (Day 0), the media were removed, cells were washed with PBS, and differentiation media with or without heparin (100 μg/ml) were added. Differentiation media consisted of 0.1 μM dexamethasone, 450 μM 3-isobutyl-1-methylxanthine, 2 μM insulin, and 1 μM rosiglitazone in DMEM supplemented with 10% FBS containing FFA and 1× Pen-Strep (Sigma). On day 3 of differentiation, cells were washed with PBS and treated with insulin media, which consisted of 2 μM insulin, and 1 μM rosiglitazone in DMEM with 10% FBS containing FFA and 1× Pen-Strep (Sigma). The cellular cholesterol and triglyceride content was determined after lysing cells in 0.1 M NaOH. Total plasma cholesterol and plasma triglyceride levels (Sekisui Diagnostics) and protein levels (BCA protein assay) were determined using commercially available kits. All procedures were approved by the University of California San Diego Institutional Animal Care and Use Committee.

Trieger G.W., Pessentheiner A.R., Purcell S.C., Green C.R., DeForest N., Willert K., Majithia A.R., Metallo C.M., Godula K, & Gordts P.L. (2023). Glycocalyx engineering with heparan sulfate mimetics attenuates Wnt activity during adipogenesis to promote glucose uptake and metabolism. The Journal of Biological Chemistry, 299(5), 104611.

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Culturing human cell lines

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K562 human immortalised chronic myelogenous leukaemia bone marrow cells were grown in IMDM + Glutamax-I (Gibco), supplemented with 10% fetal bovine serum (Sigma) and 1x Pen./Strep. (Sigma). Human cervix adenocarcinoma HeLa S3 cells were cultured in Ham’s F-12 nutrient mix with 2 mM l-Glutamine (Gibco), supplemented with 10% fetal bovine serum (Sigma) and 1x Pen./Strep. (Sigma). Human mammary epithelial MCF10a cells were cultured in DMEM/F-12 (Sigma) supplemented with 5% horse serum (Thermofisher), 20 ng/ml EGF (Sigma), 0.5 µg/ml hydrocortisone (Sigma), 100 ng/ml cholera toxin (Sigma), 10 µg/ml insulin (Sigma), 2 mM l-Glutamine (Thermofisher) and 1x Pen./Strep. (Sigma)^{26 (link)}. All cells were grown at 37^oC, 5% CO2, and were regularly tested for mycoplasma presence.

Agirre E., Oldfield A.J., Bellora N., Segelle A, & Luco R.F. (2021). Splicing-associated chromatin signatures: a combinatorial and position-dependent role for histone marks in splicing definition. Nature Communications, 12, 682.

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Osteogenic Differentiation of eMSCs on PCL Scaffolds

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To ensure effective removal of solvents all scaffolds used for in vitro cell experiments were washed for 6 h in 70 v/v% ethanol in water followed by 5 times washing with Mili-Q water for 24h. After washing, samples were sterilized for 2h under ultraviolent (UV) light and subsequently immersed in cell culture media supplemented with 1v/v% penicillin/streptomycin (Pen-Strep) (all Gibco, Thermo Fisher, USA) for 3 days to remove any remaining solvents from the printing process. Equine mesenchymal stem cells (eMSCs) were harvested from healthy bone marrow aspirates according to a protocol described elsewhere [31 (link)]. EMSCs were then first expanded 7 days in α-MEM supplemented with 10% (v/v) fetal bovine serum (FBS), 0.2 mM L-ascorbic-acid-2-phosphate (ASAP), and 1% (v/v) Pen-Strep at 37 °C in a humidified atmosphere containing 5% CO2, and then seeded (passage number = 3) onto scaffolds (PCL, MgPSr-PCL30, and MgP-PCL30) at a density of 30, 000 cells per cm². Cell-laden constructs were cultured in basal media for 7 days, then divided into two groups: samples cultured in 1) basal medium (α-MEM+10% FBS+0.2 mM ASAP+1% Pen-Strep, Sigma-Aldrich, Germany) and in 2) osteogenic medium (α-MEM+10% FBS+0.2 mM ASAP+1% Pen-Strep+ 10 nM Dexamethasone,+10 mM B-glycerophosphate, Sigma-Aldrich, Germany). Medium was changed every 3 days, and at least 3 scaffolds were tested per group.

Golafshan N., Vorndran E., Zaharievski S., Brommer H., Kadumudi F.B., Dolatshahi-Pirouz A., Gbureck U., Van Weeren R., Castilho M, & Maldaa J. (2020). Tough magnesium phosphate-based 3D-printed implants induce bone regeneration in an equine defect model. Biomaterials, 261, 120302.

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Cell Culture of HEK293T, A549, and THP1

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The human embryonic kidney epithelial cell line HEK293T (ATCC, CRL-3216), and human lung epithelial cell line A549 (ATCC, CCL-185) were cultured in Dulbecco’s Modified Eagle’s medium (DMEM; high glucose, Sigma) supplemented with 10% (v/v) FBS (Sigma) and 1% (v/v) PenStrep (Sigma) and maintained in a 5% CO₂ incubator at 37°C. THP1 (ATCC, CRL-TIB-202) cells were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (R8758, Sigma) supplemented with 10% (v/v) FBS (Sigma) and 1% (v/v) PenStrep (Sigma) and maintained in a 5% CO₂ incubator at 37°C. Each cell lines were confirmed free from mycoplasma contamination by testing with mycoplasma detection kit (Sigma).

Khan S., Shafiei M.S., Longoria C., Schoggins J., Savani R.C, & Zaki H. (2021). SARS-CoV-2 spike protein induces inflammation via TLR2-dependent activation of the NF-κB pathway. bioRxiv.

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Culturing Diverse Cell Lines for Research

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C2C12, HEK293T and RD cell lines were purchased from ATCC (Manassas, VA). JR1, Rh36, Rh30 and Rh41 cell lines were generously donated by Dr. Peter Houghton (Columbus, OH, USA). Mouse embryonic fibroblasts (MEFs) were isolated from E13.5 embryos of mixed C57BL/6 × 129/Sv 77 background (Jackson Laboratory, Maine) using the procedure approved by the Institutional Care and Use Committee (IACUC) at the American University of Beirut, and following the IACUC-approved guidelines. C2C12 cells were maintained in Dulbecco’s Modified Eagles Medium (DMEM) with 20% FBS, 1% glutamine, and 1% Pen/Strep (Sigma). Other cells were cultured in RPMI-1640 medium with 10% fetal bovine serum, 1% glutamine, and 1% Pen/Strep (Sigma). All cells maintained under standard conditions (humidified atmosphere, 95% air, 5% CO₂, 37 °C).

Ghamloush F., Ghayad S.E., Rammal G., Fahs A., Ayoub A.J., Merabi Z., Harajly M., Zalzali H, & Saab R. (2019). The PAX3-FOXO1 oncogene alters exosome miRNA content and leads to paracrine effects mediated by exosomal miR-486. Scientific Reports, 9, 14242.

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