Reagentpack

Manufactured by Lonza

Sourced in Switzerland, United States, Australia

ReagentPack is a laboratory equipment product designed for the storage and transport of reagents. It provides a secure and controlled environment to maintain the integrity of various chemical substances used in scientific research and analysis.

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Lab products found in correlation

Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions) Begm bronchial epithelial cell growth medium bulletkit, by Lonza (1 mentions) Begm bullet kit, by Lonza (1 mentions) Horse serum, by Merck Group (1 mentions) Epidermal growth factor (egf), by Merck Group (1 mentions) Dmem ham s f12, by Merck Group (1 mentions) Hydrocortisone, by Merck Group (1 mentions) Cholera toxin, by Merck Group (1 mentions) Astrocyte basal medium, by Lonza (1 mentions) Cc 2565, by Lonza (1 mentions) Singlequots kit, by Lonza (1 mentions) Saos 2, by American Type Culture Collection (1 mentions) Singlequots, by Lonza (1 mentions) Bronchial epithelial growth medium (begm), by Lonza (1 mentions) Nhbe cells, by Lonza (1 mentions)

Spelling variants of this product

ReagentPack™ Subculture Reagents (18 citations) ReagentPack™ subculturing reagents (3 citations) ReagentPack subculture kit (2 citations) ReagentPack Subculture Reagent Kit (2 citations)

8 protocols using reagentpack

Hypoxia Induction and DBP Silencing in Bladder Smooth Muscle Cells

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Primary bladder smooth muscle cells (BdSMC) were provided by Lonza and cultured using Reagent Pack (Lonza, Basel, Switzerland) at 37 °C under 5% CO₂. To induce hypoxia in cell culture, CoCl₂ (Nacalai Tesque) was used at a final concentration of 500 mM in culture medium. Cells were incubated in CoCl₂-containing medium for 4–24 h at 37 °C in an atmosphere containing 5% CO₂.
To knockdown DBP expression, transfection of siRNA targeting DBP (cat. no. SI00359751; Qiagen) and siRNA negative control (cat. no. 452002; Invitrogen, CA. USA) was performed using Lipofectamine RNAiMAX (Invitrogen) for 24 h.

Ito S., Nomura T., Ueda T., Inui S., Morioka Y., Honjo H., Fukui A., Fujihara A., Hongo F, & Ukimura O. (2021). Gene expression profiles during tissue remodeling following bladder outlet obstruction. Scientific Reports, 11, 13171.

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Primary Bronchial Epithelial Cell Culture

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Primary normal human bronchial epithelial (NHBE) and DHBE cells were purchased from Lonza (Basel, Switzerland). Cell cultures were performed according to the manufacturer’s protocol. Cells were grown in bronchial epithelial cell basal media (BEGM™ Bronchial Epithelial Cell Growth Medium BulletKit™; Lonza), supplemented with 2 mL bovine pituitary extract, 0.5 mL hydrocortisone, 0.5 mL human epidermal growth factor (hEGF), 0.5 mL epinephrine, 0.5 mL transferrin, 0.5 mL insulin, 0.5 mL retinoic acid, 0.5 mL triiodothyronine, and 0.5 mL GA1000. Cells were maintained at 37°C in a 5% CO₂ incubator and passaged with Reagent-Pack (Lonza) containing trypsin–EDTA (EDTA, trypsin neutralizing solution, and HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid]-buffered solution). Cells were harvested for NGS analysis after cultivation from primary cells for one generation.

Chang W.A., Tsai M.J., Jian S.F., Sheu C.C, & Kuo P.L. (2018). Systematic analysis of transcriptomic profiles of COPD airway epithelium using next-generation sequencing and bioinformatics. International Journal of Chronic Obstructive Pulmonary Disease, 13, 2387-2398.

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Culture and Maintenance of Primary Human Bronchial Epithelial Cells

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Primary normal human bronchial epithelial cells (NHBE) and asthmatic bronchial epithelial cells (DHBE) were purchased from Lonza Walkersville, Inc. The cell culture was performed following the manufacturer’s protocol. Cells were grown in Bronchial Epithelial Cell Basal Media (BEGM Bulletkit™; Lonza, Clonetics, San Diego, CA) supplemented with 2 ml BPE, 0.5 ml hydrocortisone, 0.5 ml hEGF, 0.5 ml epinephrine, 0.5 ml transferrin, 0.5 ml insulin, 0.5 ml retinoic acid, 0.5 ml triiodothyronine, and 0.5 ml GA-1000. Cells were maintained at 37°C in 5% CO₂ incubator, and passaged with ReagentPack™ (Lonza; Clonetics, San Diego, CA, USA) containing trypsin-EDTA, trypsin neutralizing solution, and HEPES buffered solution.

Sheu C.C., Tsai M.J., Chen F.W., Chang K.F., Chang W.A., Chong I.W., Kuo P.L, & Hsu Y.L. (2017). Identification of novel genetic regulations associated with airway epithelial homeostasis using next-generation sequencing data and bioinformatics approaches. Oncotarget, 8(47), 82674-82688.

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Culturing Diverse Breast Cell Lines

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HEK293T and the breast carcinoma cell lines BT549 and MDA-MB-231, were from the ATCC repository, thawed and cultured accordingly for fewer than 15 passages. Primary human epithelial mammary cells (HMEC) were purchased (CC-2551, Lonza, Australia), thawed and cultured using Mammary Epithelia Growth Medium and Bullet Kit™ (CC-3150, Lonza, Australia) and ReagentPack™ (CC-5034, Lonza, Australia) for fewer than 4 passages. MCF10A cells were purchased from ATCC^® repository (CRL-10317), thawed and cultured for fewer than 10 passages in 1:1 DMEM/Ham’s F12 (Sigma 51651C), 5% Horse serum (Sigma H1270), 20 ng/ml EGF (Sigma E9644), 0.5 ug/ml Hydrocortisone (Sigma H0888), 100 ng/ml Cholera Toxin (Sigma C8052), 0.25 U/ml Insulin (Protaphane Penfill, Novo Nordisk Pharmaceuticals PTY. LTD, NSW, Australia).

Brown C.Y., Dayan S., Wong S.W., Kaczmarek A., Hope C.M., Pederson S.M., Arnet V., Goodall G.J., Russell D., Sadlon T.J, & Barry S.C. (2018). FOXP3 and miR-155 cooperate to control the invasive potential of human breast cancer cells by down regulating ZEB2 independently of ZEB1. Oncotarget, 9(45), 27708-27727.

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Transduction of hABCs and hAECs for Bioluminescence

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hABCs and hAECs were grown on 75-cm² collagen-coated flasks, expanded to 75% confluency, treated with the LV-Luc vector at the chosen MOI of 10, and incubated at 37 °C and 5% CO₂ overnight in a humidified chamber. To confirm that cells expressed Luc prior to transplantation, a subset were examined by using bioluminescence imaging (IVIS Lumina XRMS, Xenogen Corporation, Alameda, CA, USA). Media was removed from the flasks, 2 mL of D-luciferin (30 mg/mL in PBS) was added, and imaging was performed. A circular region of interest (ROI) was used for quantification, and blank flasks containing no cells were imaged as controls.
On the following day, transduced cells (designated hABC-Luc and hAEC-Luc) were harvested by using HEPES-buffered saline, Trypsin/EDTA, and Trypsin neutralising solution (ReagentPack, cc-5034, Lonza) in accordance with the instructions of the manufacturer. Cells were re-suspended in PBS, cell counts were obtained, and 1.3 × 10⁵ cells per aliquot were held on ice for immediate delivery into mouse airways.

Farrow N., Cmielewski P., Donnelley M., Rout-Pitt N., Moodley Y., Bertoncello I, & Parsons D. (2018). Epithelial disruption: a new paradigm enabling human airway stem cell transplantation. Stem Cell Research & Therapy, 9, 153.

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Culturing Normal Human Astrocytes

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The Normal Human Astrocyte (NHA; Lonza, CC-2565) cell line was implemented in this study because its neuroglial lineage and species origin present a practical in vitro system for studying the effects of hydrogel biomaterials on the human brain. Furthermore, because NHA are a commercially available cell line, they can be used in follow-up and confirmatory experiments by our research team as well as those of other investigators. NHA from two different biological donors (lots #000080982, #000022529; referred to hereafter as Donor A and Donor B) were cultured according to the manufacturer’s specifications. All cytokines, growth factors, and supplements from the SingleQuots™ kit (Lonza; CC-4123) were added to Astrocyte Basal Medium (Lonza; CC-3187). We omitted the manufacturer-recommended gentamicin from cell media because the aseptic techniques used in our laboratory enable NHA culture in an antibiotic-free environment. Frozen ampules of cells were thawed and plated into four T-25 flasks (passage 0) and incubated at 37°C, 5% CO₂, 275 mOsm. Media were replenished within 24 h of thawing cells, and every 48 h that followed. NHA were subcultured by partial digestion with ReagentPack™ subculturing reagents (Lonza; CC-5034) when cultures reached 80% confluence, five days after plating (passage 1).

Knight V.B, & Serrano E.E. (2017). Hydrogel scaffolds promote neural gene expression and structural reorganization in human astrocyte cultures. PeerJ, 5, e2829.

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Culturing Human Osteosarcoma Cells on Bone Surfaces

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The human osteosarcoma cells MG63, Hos and Saos-2 were obtained from the American Type Culture Collection (Manassas, VA). MG63 and Hos cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM), while Saos-2 cells were cultured in RPMI-Medium 1640. Cell culture media and supplements for the above cell lines were purchased from Gibco (Waltham, MA). All media were supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin/100 μg/ml streptomycin, and 1% L-glutamine. Cells were maintained at 37ºC in a humidified 5% carbon dioxide atmosphere, and sub-cultured at a 1:5 ratio. Two lots (426160 and 435102) of primary human osteoblast (termed HOB1 and HOB2) were procured from Lonza (Basel, Switzerland) and cultured in OBM Basal Medium (Lonza), supplemented with SingleQuots (Lonza), at 37ºC in a humidified 5% carbon dioxide atmosphere. Sub-cultivation was carried out at 80% confluency, using the recommended ReagentPack (Lonza), following the manual´s instructions.
To study cells on mineralized or demineralized bone surfaces, cells were seeded onto the osseous surface at a density of 200,000 cells/cm² and cultivated for 72 hours. For harvest, cells were mechanically removed using a cell scraper, and collected in their appropriate medium through centrifugation (500 g for 5 minutes at RT).

Wischmann J., Lenze F., Thiel A., Bookbinder S., Querido W., Schmidt O., Burgkart R., von Eisenhart-Rothe R., Richter G.H., Pleshko N, & Mayer-Kuckuk P. (2018). Matrix mineralization controls gene expression in osteoblastic cells. Experimental cell research, 372(1), 25-34.

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Synchronization and IL-1β Treatment of Primary NHBE Cells

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Primary NHBE cells were purchased from Lonza (Walkersville, MD) and cultured as monolayers in serum-free bronchial epithelium growth medium supplemented with growth factors (BEGM, Lonza). Subculture reagents (Reagentpack, Lonza) were used according to the supplier’s instructions. Passage 3 cells were seeded in a 24-well plate (6 × 10⁵ cells/well) and allowed to attach overnight before synchronization with 50% FBS in BEGM (1 h). Cells were then treated 22 h after synchronization with 1 ng/ml IL-1β. 2 h later, RNA was extracted.

Gibbs J., Ince L., Matthews L., Mei J., Bell T., Yang N., Saer B., Begley N., Poolman T., Pariollaud M., Farrow S., Demayo F., Hussell T., Worthen G.S., Ray D, & Loudon A. (2014). An epithelial circadian clock controls pulmonary inflammation and glucocorticoid action. Nature medicine, 20(8), 919-926.

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