D1830

Manufactured by Thermo Fisher Scientific

Sourced in United States

The D1830 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It is a piece of equipment designed for use in various laboratory settings. The core function of the D1830 is to perform specific tasks or operations required in the laboratory environment. No further details or interpretations about the intended use of this product are provided.

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Lab products found in correlation

6 protocols using d1830

Fluorescent Probes and Cell Culture

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Fluo-4 (F14200), Fluo-4 AM (F14201), Dextran Texas Red, 3,000, 10,000, and 70,000 MW (D3329, D1828, and D1830, respectively) were purchased from Thermo Fisher Scientific. CT04 and CN03 were the products of Cytoskeleton Inc. Cytochalasin D (C2618), cytosine arabinofuranoside (Ara-C), poly-D-lysine, and paraformaldehyde were purchased from Millipore Sigma. NGF was purified from mouse salivary glands. Goat anti-mouse Alexa 488, 10% goat serum, and laminin (1 μg/mL) were products of Life Technologies. Mouse Tuj1 primary antibody was the product of Covance. DMEM high glucose (11,965,092), DMEM/F-12 (11,320,033), DMEM/F-12 phenol red free (21,041,025), DMEM high glucose, calcium free (21,068,028), 100X penicillin/streptomycin (10,378,016), and FBS (16,000,044) were brought from Gibco.

Yong Y., Gamage K., Cushman C., Spano A, & Deppmann C. (2020). Regulation of degenerative spheroids after injury. Scientific Reports, 10, 15472.

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Microglial Dextran Uptake Dynamics

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Primary microglia were incubated with 100 μg/ml Texas red-labelled Dextran-70 (D1830, Thermo) or Dextran-40 (D1829, Thermo) for 15 min. Then the cells were chased with fresh medium immediately for confocal imaging at 552 nm by Leica SP8.

Wang X., Chen G., Wan B., Dong Z., Xue Y., Luo Q., Wang D., Lu Y, & Zhu L. (2022). NRF1-mediated microglial activation triggers high-altitude cerebral edema. Journal of Molecular Cell Biology, 14(5), mjac036.

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Measuring Endothelial Cell Permeability

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Endothelial cell permeability was measured according to published protocols with minor modification [ 12 ]. Briefly, the human cerebral endothelial cells (5 × 10 4 /well) were seeded in the insert of a transwell (0.4 µm pore, 3413, Costar) for 5 days to generate an endothelial monolayer. Fluorescent-conjugated dextran (70 kDa, 0.5 mg/ml, D1830, Thermo Fischer Scientific, Waltham, Massachusetts, USA) was added into the inner chamber of the trans-well for 30 min. Fluorescent signals at the outer chamber (the bottom well) were measured at wavelengths of 595 and 615 nm using a plate reader (Packard, Packard Bioscience Company, Meriden, Connecticut, USA). The trans-endothelial permeability was calculated as (OD 30min -OD 0min ) experimental/(OD 30min -OD 0min ) control × 100% [ 13 ].

Teng H., Li C., Zhang Y., Lu M., Chopp M., Zhang Z.G., Melcher-Mourgas M, & Fleckenstein B. (2021). Therapeutic effect of Cerebrolysin on reducing impaired cerebral endothelial cell permeability. Neuroreport, 32(5).

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Dextran Uptake in Pericardial Nephrocytes

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Adult female fly hearts were dissected in artificial Drosophila haemolymph and were incubated with 0.1 mg/ml 10-kDa dextran (AlexaFluor568, Invitrogen) or with 0.1 mg/ml 70-kDa dextran (Thermo Fisher Scientific, D1830). After 15 min of incubation, the hearts were washed three times with PBS. Pericardial nephrocytes were then immediately mounted for confocal imaging. Zen Software was used to measure the volume and signal of fluorescent dextran for the dextran quantification. At least three nephrocytes from three different flies were analyzed. Results were analyzed using a two-tailed unpaired Student's t-test (α=0.05).

Selma-Soriano E., Llamusi B., Fernández-Costa J.M., Ozimski L.L., Artero R, & Redón J. (2020). Rabphilin involvement in filtration and molecular uptake in Drosophila nephrocytes suggests a similar role in human podocytes. Disease Models & Mechanisms, 13(9), dmm041509.

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Uptake of SOD1 Aggregates in Motor Neurons

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For flow cytometry experiments, the Islet1-GFP motor neuron reporter line was used^{34 (link)}. For all experiements, cells were incubated with 0.2 µM (monomer equivalent) of DyLight 650 labeled WT or SOD1^H46R aggregates for the indicated times. Unless otherwise stated, before the analysis, the remaining inoculum was digested with 0.25% trypsin for 5 minutes, and the cells were collected for analysis by flow cytometry. Flow cytometry data capture and cell sorting were performed on a Mo-Flo cell sorter (Beckman Colter). Analysis was performed with the FlowJo software (Flowjo LLC.). Where indicated the DyLight 650 labeled WT or SOD1^H46R aggregates were replaced with the fluid phase endocytosis marker dextran (~70,000 MW, D1830, ThermoFisher), a small hydrophilic polysaccharide, at a concentration of 2.5 mg/mL.

Benkler C., O’Neil A.L., Slepian S., Qian F., Weinreb P.H, & Rubin L.L. (2018). Aggregated SOD1 causes selective death of cultured human motor neurons. Scientific Reports, 8, 16393.

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Fluorescent Labeling of Neuronal Cells

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Fluo-4 (F14200), Fluo-4 AM (F14201), Dextran Texas Red, 3,000, 10,000, and 70,000 MW (D3329, D1828, and D1830, respectively) were purchased from Thermo Fisher Scientific. CT04 and CN03 were the products of Cytoskeleton Inc. Cytochalasin D (C2618), cytosine arabinofuranoside (Ara-C), poly-D-lysine, and paraformaldehyde were purchased from Millipore Sigma. NGF was purified from mouse salivary glands. Goat anti-mouse Alexa 488, 10% goat serum, and laminin (1 µg/mL) were products of Life Technologies. Mouse Tuj1 primary antibody was the product of Covance. DMEM high glucose (11965092), DMEM/F-12 (11320033), DMEM/F-12 phenol red free (21041025), DMEM high glucose, calcium free (21068028), 100X penicillin/streptomycin (10378016), and FBS (16000044) were brought from Gibco.

Yong Y., Gamage K.K., Cushman C., Spano A., & Deppmann C.D. (2020). Regulation of degenerative spheroids after injury.

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