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1918 fn 02m

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The 1918-FN-02M is a lab equipment product. It is designed for use in research and laboratory settings. The core function of this product is to perform specific tasks related to laboratory procedures and experimentation.

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Lab products found in correlation

Neurobasal medium, by Thermo Fisher Scientific (2 mentions) Cultrex, by R&D Systems (2 mentions) Fab1354p, by R&D Systems (2 mentions) N2 supplement, by Thermo Fisher Scientific (2 mentions) B27 supplement, by Thermo Fisher Scientific (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Celltracker green cmfda, by Thermo Fisher Scientific (2 mentions) Celltracker orange cmra, by Thermo Fisher Scientific (2 mentions) L2020, by R&D Systems (2 mentions) Lithium chloride (licl), by Merck Group (1 mentions) Lucifer yellow ch dilithium salt, by Merck Group (1 mentions) Zcl278, by Bio-Techne (1 mentions) Resveratrol, by Merck Group (1 mentions) Fnr01 a, by Cytoskeleton (1 mentions) Matrigel, by BD (1 mentions) Human igg1 fc, by R&D Systems (1 mentions) Human vegf a165, by R&D Systems (1 mentions) Ni nta bead, by Qiagen (1 mentions) Hiload 16 60 superdex 200 prep grade column, by GE Healthcare (1 mentions) Growth factor reduced matrigel, by BD (1 mentions)

10 protocols using 1918 fn 02m

1

Enrichment and Maintenance of Spinal Cord Progenitors

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Briefly, SCPs were purified with Fluorescence Activated Cell Sorting (FACS) after 21–23 days of differentiation using a PE-conjugated antibody to the alpha4 integrin, CD49d (R&D Systems, FAB1354P).41 (link) After FACS, SCPs were replated onto tissue culture plates treated with fibronectin (R&D Systems, 1918-FN-02M) and laminin (Cultrex, R&D Systems, 3400–010-1) coated plates. (Plates were coated with 1ug/mL laminin and 2ug/mL fibronectin in PBS for 24 hours). Cells were maintained in culture for up to 80 days post-FACS using Neurobasal medium (Life Technologies, 21103–049) supplemented with L-glutamine (Life Technologies, 25030–081), B-27 supplement (Life Technologies, 12587–070), N2 supplement (Life Technologies, cat. no. 17502–048), and 1% fetal bovine serum (FBS) (Hyclone, cat. no. SH30070.03).
Mukherjee-Clavin B., Mi R., Kern B., Choi I.Y., Lim H., Oh Y., Lannon B., Kim K.J., Bell S., Hur J.K., Hwang W., Hyun Che Y., Habib O., Baloh R.H., Eggan K., Brandacher G., Hoke A., Studer L., Jun Kim Y, & Lee G. (2019). Comparison of three congruent patient-specific cell types for the modelling of a human genetic Schwann-cell disorder. Nature biomedical engineering, 3(7), 571-582.
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2

Investigating Cell Adhesion Molecules

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ZCL278 was purchased from Tocris and diluted in DMSO at 50 mM. ICAM-1 recombinant protein (#ADP4-050), and fibronectin isolated from human plasma (1918-FN-02M) were purchased from R&D Systems. The type I solution of rat collagen, Lucifer Yellow CH dilithium salt, Resveratrol, and Lithium Chloride were purchased from Sigma-Merck. The recombinant human CCL2/MCP-1 Protein was obtained from R&D Systems and Miltenyi Biotec.
Ayala-Nunez N.V., Follain G., Delalande F., Hirschler A., Partiot E., Hale G.L., Bollweg B.C., Roels J., Chazal M., Bakoa F., Carocci M., Bourdoulous S., Faklaris O., Zaki S.R., Eckly A., Uring-Lambert B., Doussau F., Cianferani S., Carapito C., Jacobs F.M., Jouvenet N., Goetz J.G, & Gaudin R. (2019). Zika virus enhances monocyte adhesion and transmigration favoring viral dissemination to neural cells. Nature Communications, 10, 4430.
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3

Novec-7500 Oil Interfaces for Microfluidics

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In this study, we focused on interfaces formed with the fluorinated oil Novec-7500 (F051243; Fluorochem, UK), owing to its broad application in various microdroplet microfluidic technologies and its low cytotoxicity. To each well of 24-well plates (3526; Corning, NY, USA), 1 mL Novec-7500 was added (Fig. S1A). A 20 μg/mL fibronectin solution (1918-FN-02M; R&D Systems, Minneapolis, MN, USA) in phosphate-buffered saline (PBS) was slowly introduced into the culture vessel and incubated for 1 h at room temperature (32–37 °C), and then washed four-times with PBS. Subsequently, the surfaces were washed with E-MEM and replaced with E-MEM containing 10% FBS (173012-500ML; Sigma-Aldrich, St. Louis, MO, USA). Liquid–liquid interfaces were prepared under sterile conditions, immediately before the initiation of confluent culture.
Sonoi R, & Kamihira M. (2024). A novel strategy to facilitate uniform epithelial cell maturation using liquid–liquid interfaces. Scientific Reports, 14, 12314.
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4

Plasma and Cellular Fibronectin Protocols

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Human plasma FN (1918-FN-02M) was from R&D Systems, Minneapolis, MN, USA. Rhodamine-labelled bovine FN (FNR01-A) was from Cytoskeleton, Denver, CO, USA. Cellular ED-A FN isolated from human foreskin fibroblasts, (F2518) was from Sigma Aldrich.
Mana G., Clapero F., Panieri E., Panero V., Böttcher R.T., Tseng H.Y., Saltarin F., Astanina E., Wolanska K.I., Morgan M.R., Humphries M.J., Santoro M.M., Serini G, & Valdembri D. (2016). PPFIA1 drives active α5β1 integrin recycling and controls fibronectin fibrillogenesis and vascular morphogenesis. Nature Communications, 7, 13546.
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5

Matrigel-Based Vascular Mimicry Assay

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Cells were trypsinized, counted, washed to remove serum, and plated onto a thick (40 μl) layer of growth factor reduced Matrigel (BD Biosciences) in a 96-well plate. Cells were resuspended in 150 μl of serum-free Dulbecco’s modified Eagle’s medium for a total of 1.5 × 104 cells/well. Wells were imaged 18–24 h later at × 4 on EVOS-FL scope for the presence or absence of networks. Total network length was calculated with the neurite tracing function in the NeuronGrowth plug-in in ImageJ.59 (link) For co-culture assays, breast cancer cells and HUVECs were incubated for 30 min before trypsinization in serum-free media containing 1 μm Cell Tracker Green CMFDA or Cell Tracker Orange CMRA (Life Technologies), respectively. Cells were then plated on Matrigel in EndoGRO media (2% FBS) at a concentration of 1 × 104 HUVECs+0.5 × 104 cancer cells/well. Recombinant FN1 (1918-FN-02M, R&D Systems, Minneapolis, MN, USA) and SERPINE2 (2980-PI-010, R&D Systems) were added to VM assays at concentrations of 1 and 10 μg/ml, respectively. All VM experiments were repeated in multiple independent experiments as indicated in figure legends. Representative images are shown.
Langer E.M., Kendsersky N.D., Daniel C.J., Kuziel G.M., Pelz C., Murphy K.M., Capecchi M.R, & Sears R.C. (2017). ZEB1-repressed microRNAs inhibit autocrine signaling that promotes vascular mimicry of breast cancer cells. Oncogene, 37(8), 1005-1019.
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6

Recombinant Human WARS Isoforms Expression

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Human plasma fibronectin (R&D Systems, 1918-FN-02M, 3 µg/ml), human NRP1-Fc (R&D Systems, 10455-NI-050), human VE-Cadherin-Fc (R&D Systems, 938-VC-050) and human IgG1-Fc (R&D Systems, 110-HG-100); recombinant human VEGF-A165 (R&D Systems, 293-VE).
For the expression and purification of recombinant human WARS isoforms, DNA encoding human FL-WARS, mini-WARS, or T2-WARS with a N-terminal His6-SUMO tag was cloned into pET-28a (+) vector (Novagen). WARS expression was induced in E. coli BL21(DE3) cells with 1 mM isopropyl beta-d-thiogalactopyranoside. Following cell lysis, WARS was purified using Ni-NTA beads (Qiagen). The N-terminal His6-SUMO tag was cleaved with sumo protease ULP1. The resulting tag-free WARS proteins were further purified using HiLoad 16/60 Superdex 200 prep grade column (GE Healthcare).
Gioelli N., Neilson L.J., Wei N., Villari G., Chen W., Kuhle B., Ehling M., Maione F., Willox S., Brundu S., Avanzato D., Koulouras G., Mazzone M., Giraudo E., Yang X.L., Valdembri D., Zanivan S, & Serini G. (2022). Neuropilin 1 and its inhibitory ligand mini-tryptophanyl-tRNA synthetase inversely regulate VE-cadherin turnover and vascular permeability. Nature Communications, 13, 4188.
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7

Adhesion Molecules Regulate Sphere Formation

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Six to eight individual spheres (100 μm diameter) were placed onto pre-coated wells with 25 μg/ml laminin (Sigma #L2020), human fibronectin (FN, R&D systems #1918-FN-02 M) or human vitronectin (VN, R&D systems #2349-VN-100), containing complete media, 1 μM cytosine β-D-arabinofuranoside (Ara-C, Sigma #C1768) and either 2 μg/ml IgG isotype control (Cambridge Biosciences #400101), 0.2 μg/ml anti-ADAM10 (Millipore #AB19026), 2 μg/ml anti-ADAM17 (Calbiochem #PC491) or anti-ADAM10 and ADAM17 combined. Spheres were imaged from 0 to 45 h; sphere area was calculated using Image J freehand tool.
Siney E.J., Holden A., Casselden E., Bulstrode H., Thomas G.J, & Willaime-Morawek S. (2016). Metalloproteinases ADAM10 and ADAM17 Mediate Migration and Differentiation in Glioblastoma Sphere-Forming Cells. Molecular Neurobiology, 54(5), 3893-3905.
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8

Vasculogenic Mimicry Assay for Cancer

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Cells were trypsinized, counted, washed to remove serum, and plated onto a thick (40μL) layer of growth factor reduced matrigel (BD Biosciences, San Jose, CA, USA) in a 96-well plate. Cells were resuspended in 150μL of serum free DMEM for a total of 1.5×104 cells per well. Wells were imaged 18–24 hours later at 4× on EVOS-FL scope for the presence or absence of networks. Total network length was calculated with the neurite tracing function in the NeuronGrowth plug-in in ImageJ(59 (link)). For co-culture assays, breast cancer cells and HUVECs were incubated for 30 minutes prior to trypsinization in serum free media containing 1μM Cell Tracker Green CMFDA or Cell Tracker Orange CMRA (Life Technologies), respectively. Cells were then plated on matrigel in Endogro media (2% FBS) at a concentration of 1×104 HUVECs + 0.5×104 cancer cells per well. Recombinant FN1 (1918-FN-02M, R&D Systems) and SERPINE2 (2980-PI-010, R&D Systems) were added to VM assays at concentrations of 1μg/mL and 10μg/mL, respectively. All VM experiments were repeated in multiple independent experiments as indicated in figure legends. Representative images are shown.
Langer E.M., Kendsersky N.D., Daniel C.J., Kuziel G.M., Pelz C., Murphy K.M., Capecchi M.R, & Sears R.C. (2017). ZEB1-repressed microRNAs inhibit autocrine signaling that promotes vascular mimicry of breast cancer cells. Oncogene, 37(8), 1005-1019.
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9

Enrichment and Maintenance of Spinal Cord Progenitors

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Briefly, SCPs were purified with Fluorescence Activated Cell Sorting (FACS) after 21–23 days of differentiation using a PE-conjugated antibody to the alpha4 integrin, CD49d (R&D Systems, FAB1354P).41 (link) After FACS, SCPs were replated onto tissue culture plates treated with fibronectin (R&D Systems, 1918-FN-02M) and laminin (Cultrex, R&D Systems, 3400–010-1) coated plates. (Plates were coated with 1ug/mL laminin and 2ug/mL fibronectin in PBS for 24 hours). Cells were maintained in culture for up to 80 days post-FACS using Neurobasal medium (Life Technologies, 21103–049) supplemented with L-glutamine (Life Technologies, 25030–081), B-27 supplement (Life Technologies, 12587–070), N2 supplement (Life Technologies, cat. no. 17502–048), and 1% fetal bovine serum (FBS) (Hyclone, cat. no. SH30070.03).
Mukherjee-Clavin B., Mi R., Kern B., Choi I.Y., Lim H., Oh Y., Lannon B., Kim K.J., Bell S., Hur J.K., Hwang W., Hyun Che Y., Habib O., Baloh R.H., Eggan K., Brandacher G., Hoke A., Studer L., Jun Kim Y, & Lee G. (2019). Comparison of three congruent patient-specific cell types for the modelling of a human genetic Schwann-cell disorder. Nature biomedical engineering, 3(7), 571-582.
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10

Dissociated GSC Sphere Adhesion Assay

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GSC spheres were dissociated and passed through a 40-μm filter prior to adding into a 48-well plate pre-coated with 25 μg/ml laminin (Sigma #L2020), human fibronectin (FN, R&D systems #1918-FN-02M) or human vitronectin (VN, R&D systems #2349-VN-100) at 5 × 103 cells per well in complete media. These cultures were maintained at 37 °C, 5 % CO2 for 14 days with a 50 % media change at days 5 and 10.
Siney E.J., Holden A., Casselden E., Bulstrode H., Thomas G.J, & Willaime-Morawek S. (2016). Metalloproteinases ADAM10 and ADAM17 Mediate Migration and Differentiation in Glioblastoma Sphere-Forming Cells. Molecular Neurobiology, 54(5), 3893-3905.
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