Hypoxic glove box

Manufactured by Coy Laboratory Products

Sourced in United States

The Hypoxic Glove Box is a controlled-environment laboratory enclosure designed to maintain low oxygen levels. It provides an isolated workspace for handling and manipulating samples in a hypoxic (low oxygen) atmosphere.

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Lab products found in correlation

Teklad global rodent diet t 2018c 12, by Inotiv (1 mentions) Hepg2 cell, by American Type Culture Collection (1 mentions) Fatty acid free bsa, by Roche (1 mentions) Colorimetric assay, by Abcam (1 mentions) Penicillin, by MP Biomedicals (1 mentions) Streptomycin, by MP Biomedicals (1 mentions) Ne 1002x, by New Era Pump Systems (1 mentions)

5 protocols using hypoxic glove box

Maternal Hypoxia Impact on Fetal Development

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All experiments were conducted according to the Finnish Act on Animal Experimentation (62/2006) and approved by National Animal Experiment Board of Finland. Young (3–6 months old) female C57BL/6NCrl mice were mated under normoxic conditions (21% oxygen) overnight. The next morning (E0.5) the mice were placed either in a hypoxic chamber (Hypoxic Glove Box, Coy Laboratory Products, USA) under 15% normobaric oxygen concentration, or kept under normoxia. The dams were weighed five days a week and fed ad libitum with Teklad Global Rodent diet T.2018C.12 (Harlan Teklad, USA) during gestation.
The dams were sacrificed using CO₂ and embryos by decapitation at E9.5 or E17.5. Terminal blood and tissues were collected. Gonadal WAT, liver and placentas were weighed and tissues were snap frozen in liquid nitrogen. The embryo number was calculated, embryos stripped of fetal membranes, and the pooled embryo weight measured for each dam.

Määttä J., Sissala N., Dimova E.Y., Serpi R., Moore L.G, & Koivunen P. (2018). Hypoxia causes reductions in birth weight by altering maternal glucose and lipid metabolism. Scientific Reports, 8, 13583.

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HepG2 Cell Culture under Hypoxia

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HepG2 cells were obtained from the American Type Culture Collection (Rockville, Maryland, USA) and cultured as recommended by the supplier. Cells were maintained in minimum essential medium+Earle’s salts+l-glutamine, supplemented with 10% foetal bovine serum heat-inactivated and 1% nonessential amino acids (all from PAA Laboratories, GmBH, Cölbe, Germany). For hypoxic incubations (1% oxygen), the cells were cultured in a hypoxic glove box (COY Laboratory Products Inc., Grass Lake, Michigan USA).

Bowyer C., Lewis A.L., Lloyd A.W., Phillips G.J, & Macfarlane W.M. (2017). Hypoxia as a target for drug combination therapy of liver cancer. Anti-Cancer Drugs, 28(7), 771-780.

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Standardized Cell Culture Isotope Labeling

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All cell lines were from ATCC, were routinely passaged in Dulbecco’s Modified Eagle Medium (DMEM; Mediatech) with 25 mM glucose and 4 mM glutamine, were supplemented with 10% fetal bovine serum (HyClone), 25 IU/ml penicillin, and 25 μg/ml streptomycin (MP Biomedicals), and were split at 80% confluence. Metabolic experiments were performed in 6-cm culture dishes with 3 ml of DMEM containing 10% dialyzed serum (DFBS; HyClone). For isotope labeling experiments, glucose and/or amino acids were replaced as indicated with their U-¹³C-labeled forms (Cambridge Isotope Labs). U-¹³C-acetate was spiked into the medium to achieve indicated concentrations. U-¹³C-palmitate was complexed to fatty-acid-free BSA (Roche) in 6:1 molar ratio in a 150 mM NaCl solution by stirring overnight at 37°C and provided to cells to the indicated concentration. For all ¹³C-tracing experiments, cells were maintained in the labeled medium for 48 h unless otherwise indicated. Medium acetate was quantified by colorimetric assay (BioVision). Hypoxia experiments were performed in a hypoxic glove box (1% O₂, 5% CO₂, and 94%–94.5% N₂, at 37°C) (Coy Laboratory Products). Cells and media were equilibrated in low oxygen overnight before experiment initiation.

Kamphorst J.J., Chung M.K., Fan J, & Rabinowitz J.D. (2014). Quantitative analysis of acetyl-CoA production in hypoxic cancer cells reveals substantial contribution from acetate. Cancer & Metabolism, 2, 23.

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Continuous Flow Migration Assay

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A Boyden chamber (NeuroProbe, BY312) was modified by cutting the chamber in half, leaving the upper and lower wells separated by a filter held in place by an acetal retainer. A trimmed p200 pipette tip was inserted in the opening of the lower well, and a hole was drilled on the opposite side of the lower well so a tube could be connected to a syringe. These modifications facilitated continuous flow of media through the lower well (Fig. 1a). Migration assays were carried out in a hypoxic glove box (Coy Laboratory Products) at 37 °C, 5% CO₂, 20% O_2. Media was perfused at 0.2 mL/min with a syringe pump (New Era Pump Systems, NE-1002X). Growth media was used in both upper and lower wells. Polycarbonate membranes with 5-μm-diameter pores or 14-μm-diameter pores (NeuroProbe) were imaged using an scanning electron microscope. Migrated cells were collected and processed for sequencing every 30 min for 3–4 h (5-μm pores) or 1 h (14-μm pores). Unmigrated control cells were not exposed to the migration chamber and were processed for sequencing during the first 30 min of the migration assay. All processed time points were pooled and frozen at − 80 °C prior to RNA extraction or Hi-C library preparation.

Jacobson E.C., Perry J.K., Long D.S., Olins A.L., Olins D.E., Wright B.E., Vickers M.H, & O’Sullivan J.M. (2018). Migration through a small pore disrupts inactive chromatin organization in neutrophil-like cells. BMC Biology, 16, 142.

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Isolation and Hypoxic Treatment of Mouse and Chicken RBCs

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Mouse and chicken RBCs were isolated by differential centrifugation of whole blood at 845 x g for 3 min at 4°C. The top plasma and buffy coat layers were discarded. The packed RBCs were washed 3 times by resuspension in 3 volumes of glucose-free PBS (137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4; pH 7.4), centrifuged and re-suspended in equal volumes of PBS.
RBCs were used within 2 hours of blood collection or stored at -80°C for western blot analysis. For normoxic condition, RBCs were incubated in a humidified atmosphere containing 5% CO2 and 95% air. Hypoxia was achieved in vitro by incubation in a hypoxic glove box (Coy Laboratory Products) with a flow of a humidified mixture of 1% O2, 94% N2 and 5% CO2 at 37°C. Reagents used under hypoxic condition were prepared inside the glove box using solvents flushed with N2 for 30 minutes.

Wondimu E.T., Zhang Q., Jin Z., Fu M., Torregrossa R., Whiteman M., Yang G., Wu L, & Wang R. (2022). Effect of hydrogen sulfide on glycolysis-based energy production in mouse erythrocytes. Journal of cellular physiology, 237(1).

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