Glucose uptake of cells on Matrigel‐coated beads was measured through cellular uptake of 2‐NBDG (2‐(N‐(7‐Nitrobenz‐2‐oxa‐1,3‐diazol‐4‐yl) Amino)‐2‐Deoxyglucose) (Thermo Fisher). Matrigel‐coated beads for assay were washed twice with DMEM without glucose, incubated with 400µM 2‐NBDG for 30 minutes in presence or absence of 1µM Dapagliflozin inhibitor. Beads were washed twice with DMEM without glucose before cells were harvested from beads using Trypsin/EDTA. Uptake of 2‐NBDG was measured using MACSQuant Analyzer (Miltenyi Biotec) and data analyzed with FlowJo software.
2 deoxyglucose
Manufactured by Thermo Fisher Scientific
2-deoxyglucose is a synthetic glucose analog that is used as a tracer in various research and diagnostic applications. It is structurally similar to glucose, but with a hydrogen atom instead of a hydroxyl group at the 2-carbon position. This modification allows 2-deoxyglucose to be transported into cells and phosphorylated by hexokinase, but it cannot be further metabolized, leading to its accumulation within cells.
Automatically generated - may contain errors
Lab products found in correlation
Chicken ovalbumin, by Merck Group (2 mentions)
Interleukin 4 (il 4), by BD (2 mentions)
D glucose, by Thermo Fisher Scientific (2 mentions)
4 hydroxytamoxifen, by Merck Group (2 mentions)
Lipopolysaccharides (lps), by Merck Group (2 mentions)
Tamoxifen, by Merck Group (2 mentions)
Ifn γ, by BD (2 mentions)
Tgf β, by R&D Systems (2 mentions)
Oligomycin, by Merck Group (2 mentions)
2 nbdg, by Thermo Fisher Scientific (2 mentions)
Macsquant analyzer, by Miltenyi Biotec (1 mentions)
Insulin, by Merck Group (1 mentions)
Glucose test kit, by Applygen (1 mentions)
5 protocols using 2 deoxyglucose
1
Cellular Glucose Uptake on Matrigel Beads
2
Immunological Reagents for Cell Culture and Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IFN-γ, IL-4 and mAb (purified, biotinylated, or fluorophore-conjugated) were from BD Pharmingen or Tonbo Biosciences (San Diego CA) unless otherwise indicated. IL-5 was from Peprotech (Rocky Hill NJ), TGF-β and BAFF were from R&D Systems (Minneapolis, MN). NP-BSA (for capture ELISA), NP-OVA, and NP-O-succinimide were obtained from Biosearch (Novato CA). SRBC (sheep red blood cells), D-glucose, and 2-deoxyglucose were from Thermo Fisher Scientific (Waltham MA). Tamoxifen, 4-hydroxy-Tamoxifen, chicken ovalbumin, all-trans retinoic acid and LPS were from Sigma-Aldrich Chemicals (St. Louis MO). DMOG and oligomycin were from EMD Millipore (Billerica, MA). Fluorescent proteins APC and rPE (Prozyme, Haward CA) were used for conjugation reactions with NP-O-succinimide to generate fluor-conjugated NP.
3
Quantifying Cellular Glucose Uptake
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To quantify glucose uptake the commercially available fluorescent glucose probe 2-NBDG (2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (#N13195, Thermo Fisher Scientific) was used according to the manufacture instruction. Briefly, cells were washed with warm (37 °C) PBS before incubation with the compound for 20 min at 37 °C. The cells were then washed with cold (4 °C) PBS to decrease metabolic activity before taking pictures with a Nikon Eclipse TE300 inverted microscope (Bergman Labora). For each condition cell were sided in 4 replicas and a minimum of 4 images were taken per well. All experiments were carried out for at least 3 independent times. Images were analyzed using Image J software (https://imagej.nih.gov/ij/ ).
4
Immunological Reagents for Cell Culture and Assays
5
Insulin Resistance in HepG2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The IR of HepG2 cells was established based on the study of Zhang et al. [24 (link)] with modification. Briefly, cells were cultured and serum-starved for 2 h in 96-well plates, and the medium was replaced by serum-free DMEM supplemented with 10−5, 10−6, 10−7, and 10−8 mol/L insulin (Sigma) for 36 h. Subsequently, the medium was collected and the glucose content was measured using a glucose test kit (Applygen, China). In addition, glucose uptake was measured using the fluorescent glucose analog, 2-NBDG (2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose; Thermo Fisher Scientific). In this study, the glucose consumption and uptake of the 10−6 mol/L insulin-treated groups significantly decreased, indicating that the HepG2-IR model was established successfully (Figure 2S ).
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!