Zeocin

Manufactured by InvivoGen

Sourced in United States, France, Japan, Germany, China

Zeocin is a selection antibiotic used for the stable transfection of mammalian and bacterial cells. It functions by inhibiting protein synthesis, leading to cell death in non-resistant cells.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Zeocin (ant-zn-1, (3 citations) Zeocin antibiotic (3 citations) Zeocin (ant-zn-05, (2 citations)

408 protocols using zeocin

HEK 293 Cell-Based Receptor Activation Assay

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

HEK 293 cells (Invivogen, San Diego, CA, USA) were initially cultured in DMEM with 10% (v/v) fetal bovine serum, 100 U/mL penicillin, 100 mg/mL streptomycin and 100 mg/mL Normocin™ (Invivogen, San Diego, CA, USA). After the 3rd passage, selective antibiotics were added to the growth medium as required: (1) null: 100 μg/mL of Zeocin^TM (Invivogen, San Diego, USA); (2) TLR2 and TLR4: 1X HEK-Blue^TM selection (Invivogen, San Diego, USA); (3) NOD2: 30 μg/mL of blasticidin (Invivogen, San Diego, USA) and 100 μg/mL of Zeocin (Invivogen, San Diego, USA). The cell cultures were renewed with the use of PBS and without centrifuging when confluency reached 80% of the bottle. Finally, the cells were kept at 37 °C with 5% CO₂ and appropriate humidity.
On the day of the stimulation, cells were detached with PBS, counted and dissolved in HEK-Blue^TM Detection (Invivogen, San Diego, CA, USA) (~140,000 cells/mL). Then, 190 μL of the cell suspension was added to each well in the 96-well plate and stimulated with the studied samples (10 µL). Since HEK 293 cells are co-transfected with SEAP (secreted embryonic alkaline phosphatase) genes, stimulation followed by the activation of analyzed receptors gave a colorimetric reaction that was developing over time and which was measured with an absorbance read at 490, 510 and 530 nm.

Kazana W., Jakubczyk D., Pacyga-Prus K., Leszczyńska K., Górska S., Siednienko J., Macała J., Piechowiak G, & Zabłocka A. (2022). A Novel Mechanism of Macrophage Activation by the Natural Yolkin Polypeptide Complex from Egg Yolk. International Journal of Molecular Sciences, 23(6), 3125.

+ Open protocol

+ Expand

STING Binding Assay for Mouse and Human Variants

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

Mouse STING binding kits, human STING WT binding kits, human AQ STING binding kits, human H232 STING binding kits were purchased from Cisbio. 293T mSTING (ISG/KI-IFNb) cells, 293T hSTING-R232 (ISG/KI-IFNb) cells, THP1-KI-hSTING-R232 cells, THP1-KO-STING cells were purchased from Invivogen. 293T mSTING cells and 293T hSTING-R232 cells were cultured using DMEM (Gibco, Waltham, MA, USA), 2 mM L-glutamine (Sigma-Aldrich, St. Louis, MO, USA), 4.5 g/L glucose (Sigma-Aldrich), 10% FBS (Gibco), Pen-Strep (100 U/mL-100 μM) (Gibco), 100 μM normocin (Invivogen, San Diego, CA, USA), and supplemented with selective antibiotics (Invivogen) blasticidin (10 µM), hygromycin (100 µM) and zeocin (100 µM). THP1-KO-STING cells and THP1-KI-hSTING-R232 cells were cultured using RPMI-1640 (Gibco), 2 mM L-glutamine (Sigma-Aldrich), 25 mM HEPES (Sigma-Aldrich), 10% heat-inactivated FBS (Gibco), Pen-Strep (100 U/mL-100 μM) (Gibco), 100 μM normocin (Invivogen), and supplemented with selective antibiotics (Invivogen) blasticidin (10 μM) and zeocin (100 μM). QUANTI-Blue solution, QUANTI-Luc, 2′,3′-cGAMP were purchased from Invivogen. H151 was obtained from Beijing Innochem Science and Technology Co., Ltd. (Beijing, China). CellTiter-Glo luminescent cell viability assay kit was obtained from Promega.

Chang J., Hou S., Yan X., Li W, & Xiao J. (2023). Discovery of Novel STING Inhibitors Based on the Structure of the Mouse STING Agonist DMXAA. Molecules, 28(7), 2906.

+ Open protocol

+ Expand

Macrophage Differentiation and Stimulation

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

THP-1 cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and were cultured in RPMI 1640 medium (Gibco by Life Technologies, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS) (Hyclone, Logan, UT). THP-1 XBlue cells were purchased from InvivoGen (San Diego, CA). Zeocin (200 μg/ml; InvivoGen) selection antibiotic was added to Zeocin-resistant THP-1 XBlue cells in culture. THP-1 and THP-1 XBlue cells were differentiated into macrophage-like cells by culturing cells in RPMI medium +10% FBS supplemented with phorbol 12-myristate 13-acetate (PMA, 10 ng/ml; BioShop Canada Inc., Burlington, ON) for 48 hours, followed by a wash with RPMI + 10% FBS and subsequent 48 hour incubation in fresh culture medium. Recombinant human IL-27 was purchased from R&D Systems (Minneapolis, MN). LPS from Escherichia coli 0111:B4 (LPS-E) and LPS from Salmonella enterica serotype Typhimurium (S. typhimurium) (LPS-S) were purchased from Sigma-Aldrich (St. Louis, MO). Flagellin (Ultrapure) from S. typhimurium was purchased from InvivoGen.

Petes C., Odoardi N., Plater S.M., Martin N.L, & Gee K. (2018). IL-27 amplifies cytokine responses to Gram-negative bacterial products and Salmonella typhimurium infection. Scientific Reports, 8, 13704.

+ Open protocol

+ Expand

Zika Virus ZIKV Infection Protocol

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

The clinical isolate PF-25013-18 (PF13) of ZIKV has been previously described [10 (link)]. Cell lines used in this study included the A549-Dual™cell line (InvivoGen, a549d-nfis), referred to hereafter as “A549 cells”, and the HEK-Blue™ IFN-α/β cell line (InvivoGen, San Diego, CA, USA) which possesses the HEK-293A backbone and is referred to hereafter as “HEK-293A cells”. Both cell lines were cultured at 37 °C with 5% CO₂ in MEM Eagle medium, supplemented with 10% heat inactivated fetal bovine serum and 2 mmol·L⁻¹l-Glutamine, 1 mmol·L⁻¹ sodium pyruvate, 100 U·mL⁻¹ of penicillin, 0.1 mg·mL⁻¹ of streptomycin and 0.5 µg·mL⁻¹ of fungizone (PAN Biotech, Aidenbach, Germany). Additionally, A549 cellular growth medium was supplemented with 10 µg·mL⁻¹ blasticidin and 100 μg·mL⁻¹ zeocin (InvivoGen) and HEK-293A cell growth medium was supplemented with 30 µg·mL⁻¹ blasticidin and 100 μg·mL⁻¹ zeocin (InvivoGen). Cells were harvested and stored as frozen pellets for further protein or mRNA analysis.

El Kalamouni C., Frumence E., Bos S., Turpin J., Nativel B., Harrabi W., Wilkinson D.A., Meilhac O., Gadea G., Desprès P., Krejbich-Trotot P, & Viranaïcken W. (2018). Subversion of the Heme Oxygenase-1 Antiviral Activity by Zika Virus. Viruses, 11(1), 2.

+ Open protocol

+ Expand

Cell Culture and Maintenance of Immune Cell Lines

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

Parental THP-1 cells and THP1-XBlue^™-CD14 reporter cells (carrying an NF-κB/AP-1-inducible SEAP reporter construct) were obtained from InvivoGen (USA) and cultured in RPMI medium (GE Healthcare, USA) supplemented with 10% fetal calf serum (FCS, Thermo Scientific, USA), 50 U/ml penicillin, 50 μg/ml streptomycin, 2 mM glutamine, and 0.1 M NaHCO3 (all PanEco, Russia) at 37°C with 5% CO₂. 200 μg/ml Zeocin, and 250 μg/ml G418 (both InvivoGen, USA) was included in the culture medium for the THP1-XBlue^™-CD14 reporter cell line.
HEK-Blue-hTLR4, HEK-Blue-hNOD2 and control HEK-Blue-Null2 cells were obtained from Invivogen (USA) and maintained in DMEM medium (GE Healthcare, USA) supplemented with 10% fetal calf serum (Thermo scientific, USA), 50 U/ml penicillin, 50 μg/ml streptomycin, 2 mM glutamine, 0.1 M NaHCO3 (all PanEco, Russia), and 200 μg/ml Zeocin (InvivoGen, USA) at 37°C with 5% CO₂.

Tukhvatulin A.I., Dzharullaeva A.S., Tukhvatulina N.M., Shcheblyakov D.V., Shmarov M.M., Dolzhikova I.V., Stanhope-Baker P., Naroditsky B.S., Gudkov A.V., Logunov D.Y, & Gintsburg A.L. (2016). Powerful Complex Immunoadjuvant Based on Synergistic Effect of Combined TLR4 and NOD2 Activation Significantly Enhances Magnitude of Humoral and Cellular Adaptive Immune Responses. PLoS ONE, 11(5), e0155650.

+ Open protocol

+ Expand

Lentiviral Transduction of Jurkat T Cells

Cited 1 time

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

A suspension of Jurkat T cells was prepared in fresh RPMI medium with 10% FBS at a concentration of 2 × 10⁶ cells/mL containing 16 µg/mL of polybrene (Sigma Aldrich, St. Louis, MO, USA, H9268) and plated in 24-well plates at 500 µL/well. Serial dilutions of lentiviral supernatant were prepared in fresh medium and added to each well at 500 µL/well, resulting in a final concentration of polybrene of 8 µg/mL [38 (link)]. Cells were incubated at 37 °C; 24 h later, another 1000 µL of fresh medium was added to each well. At 48 h post-transduction, half of the cells were analyzed for transduction efficiency based on GFP and CAR expression at the cell surface as measured by flow cytometry. The remaining cells were recovered by centrifugation and resuspended in fresh medium containing 200 µg/mL of zeocin (Invivogen, San Diego, CA, USA, #ant-zn) at a concentration of 0.2 × 10⁶ cells/mL. Cells were kept under zeocin selection for 3 to 4 weeks with regular medium exchange and cell dilution, until reaching 1 × 10⁶ cells/mL. Thereafter, CAR-Jurkat T cells were reanalyzed for CAR expression by flow cytometry and Western blotting.

Silva G., Rodrigues A.F., Ferreira S., Matos C., Eleutério R.P., Marques G., Kucheryava K., Lemos A.R., Sousa P.M., Castro R., Barbas A., Simão D, & Alves P.M. (2023). Novel scFv against Notch Ligand JAG1 Suitable for Development of Cell Therapies toward JAG1-Positive Tumors. Biomolecules, 13(3), 459.

+ Open protocol

+ Expand

Cell Line Culture and Maintenance Conditions

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

Cell lines were kept in a humidified atmosphere with 5% CO₂ at 37°C. All media contained 10% (v/v) fetal bovine serum (FBS, PAN biotech), penicillin (100 U/ml) and streptomycin (100 μg/ml). For culture of stable mGPR17-HEK293, rGPR17-HEK293 and hGPR17–1321N1 Dulbecco’s modified Eagle’s medium (DMEM) was supplemented with G418 (500 μg/ml or 800 μg/ml) (InvivoGen), for 3HA-hGPR17-HEK293 (later named hGPR17-HEK293) with zeocin (56 μg/ml) (InvivoGen), for 3HA-hGPR17-Rluc / GFP²-β-arrestin2-HEK293 with zeocin (56 μg/ml) and G418 (500 μg/ml).
Flp-In™ T-REx CHO cells stably expressing hGPR17 (hGPR17-CHO-FITR) were maintained in DMEM with nutrient mixture F-12 (DMEM/F12) supplemented with hygromycin B (500 μg/ml) and blasticidin (30 μg/ml). Expression from the Flp-In locus was induced by treatment with doxycycline (1 μg/ml) for 16–20 h. mGPR17-CHO-Flp-In and rGPR17-CHO-Flp-In were cultivated in Ham’s F-12 nutrient mix with GlutaMAX and 600 mg/ml hygromycin B. For hM1-CHO cells, G418 (200 μg/ml) was added to Ham’s F-12 nutrient mix with GlutaMAX.

Merten N., Fischer J., Simon K., Zhang L., Schröder R., Peters L., Letombe A.G., Hennen S., Schrage R., Bödefeld T., Vermeiren C., Gillard M., Mohr K., Lu Q.R., Brüstle O., Gomeza J, & Kostenis E. (2018). Repurposing HAMI3379 to Block GPR17 and Promote Rodent and Human Oligodendrocyte Differentiation. Cell chemical biology, 25(6), 775-786.e5.

+ Open protocol

+ Expand

HEK293 Stable Cell Line Generation

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

Wild-type HEK293 T-Rex Flp-In cells (Thermo Fischer Scientific) were grown in Dulbecco’s modified Eagle’s medium (DMEM) (Thermo Fisher Scientific) supplemented with 10% (v/v) fetal bovine serum (FBS) (Thermo Fisher Scientific), 100 U ml⁻¹ Penicillin–Streptomycin (Thermo Fisher Scientific), 100 µg ml⁻¹ zeocin (InvivoGen), and 10 µg ml⁻¹ blasticidin (InvivoGen). Originating from these cells, transgenic cell lines with stable integration of constructs were generated by co-transfection of cloned FRT-plasmids (this study, see section “Plasmids”) with the Flp recombinase expression vector pOG44 (Thermo Fisher Scientific) as previously described^{62 (link)}. Positive clones of these cell lines were selected and cultured in the same media as described above except using 100 µg ml⁻¹ hygromycin (InvivoGen) instead of zeocin.

Sauer M., Juranek S.A., Marks J., De Magis A., Kazemier H.G., Hilbig D., Benhalevy D., Wang X., Hafner M, & Paeschke K. (2019). DHX36 prevents the accumulation of translationally inactive mRNAs with G4-structures in untranslated regions. Nature Communications, 10, 2421.

+ Open protocol

+ Expand

THP-1 Cell Culture and Maintenance

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

Human monocytic THP-1 cells were purchased from American Type Culture Collection (ATCC) and grown in RPMI-1640 culture medium (Gibco, Life Technologies, Grand Island, USA) supplemented with 10% fetal bovine serum (Gibco, Life Technologies, Grand Island, NY, USA), 2 mM glutamine (Gibco, Invitrogen, Grand Island, NY, USA), 1 mM sodium pyruvate, 10 mM HEPES, 100 ug/ml Normocin, 50 U/ml penicillin and 50 μg/ml streptomycin (P/S; (Gibco, Invitrogen, Grand Island, NY, USA). They were incubated at 37°C (with humidity) in 5% CO₂. TLR4-/- THP1 cells were purchased from InvivoGen (San Diego, CA, USA) and cultured in a complete RPMI medium containing Blasticidin and Zeocin antibiotics. THP1-defMyD (MyD88-/-) cells were purchased from InvivoGen (San Diego, CA, USA) and cultured in a complete RPMI medium containing Zeocin (200 μg/ml) and HygroGold (100 μg/ml). THP1 with control plasmid was purchased from InvivoGen (San Diego, CA, USA) and cultured in a complete RPMI medium containing Zeocin (200 μg/ml). Before stimulation, THP-1 cells were transferred into a normal medium.

Al-Rashed F., Haddad D., Al Madhoun A., Sindhu S., Jacob T., Kochumon S., Obeid L.M., Al-Mulla F., Hannun Y.A, & Ahmad R. (2023). ACSL1 is a key regulator of inflammatory and macrophage foaming induced by short-term palmitate exposure or acute high-fat feeding. iScience, 26(7), 107145.

+ Open protocol

+ Expand

Sertoli Cell NFkB Activation Assay

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

The pNifty2-SEAP plasmid (Invivogen) was transfected using Attractene reagent (Qiagen) for 48 hours in 1.10⁵ 15 P-1 Sertoli cells. Following 72 hours, the stimulated Sertoli cells were selected with Zeocin (250 μg/mL, Invivogen) for 5 days and subsequently cultured in the presence of 100 μg/mL Zeocin thereafter. Upon challenge with iE-DAP, MDP, or both, in separate experiments, nuclear factor kappa B (NFκB) activation was monitored live via the secreted alkaline phosphatase (SEAP) release in collected cultivation media using a colorimetric detection (Invivogen), at 405 nm (BMC FLOWStar Optima reader).

Hayrabedyan S., Todorova K., Jabeen A., Metodieva G., Toshkov S., Metodiev M.V., Mincheff M, & Fernández N. (2016). Sertoli cells have a functional NALP3 inflammasome that can modulate autophagy and cytokine production. Scientific Reports, 6, 18896.

+ 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!