2nbdg fluorescent glucose

Manufactured by Thermo Fisher Scientific

2NBDG is a fluorescent glucose analog that can be used to measure glucose uptake in cells. It functions as a fluorescent tracer to monitor cellular glucose metabolism.

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Lab products found in correlation

Mouse serum igg, by Merck Group (1 mentions) Facscalibur, by BD (1 mentions) Cytofix cytoperm kit, by BD (1 mentions)

2 protocols using 2nbdg fluorescent glucose

Immunophenotyping and Functional Assays for Macrophages

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For analysis of receptor expression, after removal from culture plates, cells were incubated with the primary antibody for 30 min at 0 °C. Primary antibodies: anti-mouse MHCII I-A/I-E (eBioscience); anti-mouse CD301 (AbD Serotec). Phagocytosis analysis was performed using 2 μm latex beads (Sigma). The beads were opsonized for 30 min at 37 °C with mouse serum IgG (Sigma) before adding to the cells and incubating for 30 min at 37 °C. Glucose uptake was assessed using 2NBDG fluorescent glucose (Invitrogen) [50 μM] for 30 min at 37 °C or 0 °C. Samples were analysed on a FACScalibur (BD). Granularity was detected in the SSC channel, and fluorescence was detected using an argon laser. Sorting of RAW264.7 cells for determination of cell doublets was performed using a FACSAria II.

Beirão B.C., Raposo T., Pang L.Y, & Argyle D.J. (2015). Canine mammary cancer cells direct macrophages toward an intermediate activation state between M1/M2. BMC Veterinary Research, 11, 151.

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Cardiomyocyte Swelling and Glucose Metabolism

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Transfected cardiomyocytes were incubated with hypotonic Krebs solution as previously described [33 (link)–35 (link)] for 40 minutes and changes in cell size, glucose uptake, and IRS-1 levels were monitored by flow cytometry. Glucose uptake was determined by adding 2-NBDG fluorescent glucose (10 μM; Invitrogen, CA) plus insulin (100 nM) for 45 minutes at 37°C with 5% CO₂. IRS-1 levels were analyzed by permeabilizing cells using BD Cytofix/Cytoperm Kit and then incubating with an anti-IRS1-antibody for 1 hour, followed by incubation with a FITC-secondary antibody. For analysis of losartan effects, transfected cells were incubated with the drug (10⁻⁷ M) for 24 hours prior to swelling and glucose uptake and IRS-1 levels were analyzed by flow cytometry.

Gerena Y., Lozada J.G., Collazo B.J., Méndez-Álvarez J., Méndez-Estrada J, & De Mello W. (2017). Losartan counteracts the effects of cardiomyocyte swelling on glucose uptake and insulin receptor substrate-1 levels. Peptides, 96, 38-43.

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