SF268 cells were plated in Corning 35-mm dishes at a concentration of 2 × 104 cells/mL in complete media. Hypoxia was induced using a STEMCELL technologies hypoxia chamber incubator (Catalog #27310) with mixed gas (1% oxygen, 5% CO2, 94% nitrogen). The chamber was purged with hypoxic gas for 15 min to fill the chamber. The cells were kept in the chamber for 24 or 48 h before being lysed.
Hypoxia incubator chamber
Manufactured by STEMCELL
Sourced in Canada, United States, France, Germany
The Hypoxia Incubator Chamber is a laboratory equipment designed to provide a controlled low-oxygen environment for cell culture and experimentation. It maintains a specified oxygen concentration within the chamber, allowing researchers to study cellular responses to hypoxic conditions.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Quantitect reverse transcription kit, by Qiagen (2 mentions)
Neurobasal medium, by Thermo Fisher Scientific (2 mentions)
Mcf 7, by American Type Culture Collection (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Cocl2, by Merck Group (2 mentions)
Penicillin streptomycin, by Corning (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
P7405, by Cellvis (2 mentions)
35 mm dishes, by Corning (1 mentions)
Fgf 1, by Thermo Fisher Scientific (1 mentions)
Low glucose dmem, by Thermo Fisher Scientific (1 mentions)
Irdye 800cw goat anti mouse igg, by LI COR (1 mentions)
Celltiter glo luminescent cell viability assay kit, by Promega (1 mentions)
Irdye 800cw goat anti rabbit igg, by LI COR (1 mentions)
Ros assay kit, by Beyotime (1 mentions)
Anti lc3b antibody, by Cell Signaling Technology (1 mentions)
68 protocols using hypoxia incubator chamber
1
Hypoxia Induction in SF268 Cells
2
Isolation and Preconditioning of Human MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bone marrow aspirates are collected from fully informed healthy human volunteer donors who provided written consent. The healthy volunteer donors are recruited from Mater Private Hospital, Brisbane, Australia. Ethical approval is granted through the Mater Health Services Human Research Ethics Committee and ratified by the Queensland University of Technology Human Ethics Committee (number: 1000000938). Human MSCs are isolated from bone marrow aspirates, cultured and characterized as we previously described (Parekkadan & Milwid, 2010 (link); Squillaro, Peluso & Galderisi, 2016 (link)). All cells are cultured in monolayer using expansion media formulated from low glucose DMEM (ThermoFischer) supplemented with 10% fetal bovine serum (FBS; Thermo Fisher, Waltham, MA, USA) and 10 ng/mL FGF-1 (Peprotech). All experiments involving MSCs are performed at passage 4-8, tested negative for mycoplasma contamination, and <80% confluence. MSCs are cultured in a hypoxia chamber incubator (catalog No. 27310; StemCell Technologies, Vancouver, BC, Canada) at 37 °C in 3% O2, 5% CO2 and 92% N2 for 24 h, and these MSCs are named as hypoxia-preconditioned MSCs. MSCs cultured for 24 h in 95% air and 5% CO2 are used as a control.
3
Hypoxia-Induced Autophagy Modulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hypoxia Incubator Chamber (Stemcell Technologies, Vancouver, Canada 27310), Single Flow Meter (Stemcell Technologies, 27311), hypoxic gas (3% O2, 5% CO2, and 92% N2, Beijing Yongsheng Gas Technology Limited Company, Beijing, China), H2O2 (Sigma-Aldrich, St. Louis, MO, USA H1009), rapamycin (Sigma-Aldrich, V900930), 3-methyxanthine (3-MA; Sigma-Aldrich, M9281), anti-β-actin antibody (TransGen Biotech, Beijing, China HC201-01), anti-LC3B antibody (Cell Signaling Technology, Inc., Danvers, MA, USA 3868), anti-Beclin 1 antibody (Cell Signaling, 3495), anti-p53 antibody (Cell Signaling, 2524), IRDye® 800CW Goat anti-rabbit IgG (LI-COR, Lincoln, NE, USA 926-32211), IRDye® 800CW Goat anti-mouse IgG (LI-COR, 926-32210), CellTiter-Glo® Luminescent Cell Viability Assay Kit (Promega Corporation, Madison, WI, USA G7571), Dulbecco’s modified Eagle’s medium (DMEM, Thermo Fisher Scientific, Waltham, MA, USA 11995073), monodansylcadaverine (MDC; Sigma-Aldrich, D4008), and ROS Assay Kit (Beyotime, Shanghai, China S0033) were used.
4
Exosome-mediated protection against hypoxia-induced cardiomyocyte apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HL-1 cardiomyocytes were seeded at a density of 6 × 104/well in six-well culture dishes and allowed to reach ~80% confluence. Cells were pre-treated for 48 h with hCPC-exosomes (100 μg) isolated from culture medium as previously reported by us21 (link). HL-1 cells treated with different hCPC-exosomes were randomly exposed for 24 h to severe hypoxia (1–2% O2) using a hypoxia incubator chamber (STEMCELL Technologies Inc., Canada), a well-established model of cardiomyocytes damage following myocardial infarction27 (link), or to normoxia (21%O2) as control condition. Detection of TUNEL staining by fluorescence microscopy (in situ cell death detection kit, Roche Diagnostic Corporation, Indianapolis, IN, USA)65 (link) and measurement of cleaved caspase-3 levels by Western blot assay66 (link) were performed to evaluate the magnitude of hypoxia-induced apoptosis of HL-1 cardiomyocytes. All measurements were performed in triplicate.
5
Anoxic Stress Response in C. elegans
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All C. elegans strains [N2(WT), egl-9(sa307), egl-9(sa307) hif-1(ia04), and hir-1(tm4098)] were maintained at 20°C before RNA extraction. For anoxic stress, we placed N2 animals into a hypoxia incubator chamber (Applied StemCell) with constant nitrogen delivering for 2 hours before lysis. One microgram of total RNA from each sample was purified by the RNeasy Mini Kit from Qiagen and used for sequencing library construction. The NEBNext rRNA Depletion Kit, Agencourt RNAClean XP Beads (Beckman Coulter), NEBNext Ultra Downloaded from https://www.science.org on September 09, 2024
Directional RNA Library Prep Kit (Illumina), Agencourt AMPure XP (Beckman Coulter), and NEBNext Multiplex Oligos (Illumina) were used to prepare sequencing libraries, as per the manufacturers' instruction. The Q5 Hot Start HiFi PCR Master Mix was used for PCR enrichment of the adaptorligated DNA. The libraries were submitted to 100 base pair (bp)-pairedend highthroughput se quencing using HiSeq3000 by the Center for Advanced Technology of the University of California, San Francisco.
Directional RNA Library Prep Kit (Illumina), Agencourt AMPure XP (Beckman Coulter), and NEBNext Multiplex Oligos (Illumina) were used to prepare sequencing libraries, as per the manufacturers' instruction. The Q5 Hot Start HiFi PCR Master Mix was used for PCR enrichment of the adaptorligated DNA. The libraries were submitted to 100 base pair (bp)-pairedend highthroughput se quencing using HiSeq3000 by the Center for Advanced Technology of the University of California, San Francisco.
6
Stress Assays in C. elegans
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In the hypoxia stress assay, animals were placed on the NGM plate with the lid and put into a hypoxic chamber containing a plate with water to maintain normal humidity. A hypoxia chamber with ProOx 110 oxygen controller (BioSpherix) was used to induce hypoxic stress with 0.5 to 21% oxygen concentrations. A hypoxia incubator chamber (Applied StemCell) with constant nitrogen flow delivery was used to achieve severe hypoxia with nearly 0% oxygen (anoxia stress). In the hydrogen peroxide stress assay, animals were placed on the plate with 10 mM peroxide in the NGM and observed in the 1 to 48hour intervals for GFP activation. To determine the effects of HIFactivating compounds on comt-5p::GFP, 5 mM CoCl 2 and 5 mM KCN containing NGM plates were used. To measure hydrogen sulfide, 0.1 mg of NaHS powder was placed onto an NGM agar plate (10 ml of agar) with a lid sealed by parafilm to prevent leaking of the released H 2 S gas. To determine the effects of inhibitors of oxidative phosphorylation on comt-5p::GFP, 1 mM rotenone, 1 mM oligomycin, and 1 mM FCCP containing NGM plates were used. Animals were screened for GFP induction in the subsequent 1 to 48 hours and collected after 2 hours of exposure for qRTPCR analysis.
7
Umbilical Cord-Derived Endothelial Cell Isolation and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immediately after delivery, umbilical cords were transported on ice from the maternity ward to the laboratory. HUAEC were separated as described previously [51 (link)]. RNA was extracted using TRIzol (Life Technologies) and quantified at 260 nm. RNA integrity was assessed by agarose gel electrophoresis. For cDNA synthesis, we used 500 ng RNA per sample using the QuantiTect Reverse Transcription kit (Qiagen). DNA was isolated by phenol:chloroform:isoamylic alcohol extraction. For in vitro experiments, HUAEC (Promocell) were cultivated upon manufacturer’s recommendations. For HDAC inhibition, HUAEC were treated with 1-μM TSA. Hypoxia experiments were performed using a hypoxia incubator chamber (STEMCELL Technologies, Grenoble, France) exposing cells to a ppO2 of 0.1 bar corresponding to an oxygen fraction of 10% for 24 h. Control experiments were performed under normoxic conditions (ppO2 = 0.21 bar).
8
Hypoxic Cell Culture Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cancer cell hypoxia model was created based upon the hypoxic microenvironment (5% O2). In this study, the Hypoxia Incubator Chamber (Cat#27310; STEMCELL TECHNOLOGIES) for generation of a hypoxic environment for cell is a self-contained and sealed chamber that fits inside existing laboratory incubators. The chambers have an integrated stacking feature for storage during or after experimentation.
9
Ozone Treatment on Pancreatic Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PANC-1 and MiaPaCa-2 cell lines were seeded on 96-well culture plates, or 12-well plates at a density of 3.0 × 104 cells/mL. The cells were pre-cultured in normoxia for 24 h. Subsequently, the culture plates were exposed twice to O2/O3 treatments for 30 min in a Hypoxia Incubator Chamber (Stemcell Technology, Vancouver, BC, Canada), by injecting O2/O3 until chamber saturation, using a Midi Ozon Active machine (Medica s.r.l., Bologna, Italy).
After treatment, the cell plates were replaced in the incubator at normoxia condition (37 °C with 5% CO2 and 95% humidity) for 6 h and then, the O2/O3 treatment was repeated as described. After that, the cells plates were maintained in normoxia for 24 up to 72 h, before performing the experiments.
After treatment, the cell plates were replaced in the incubator at normoxia condition (37 °C with 5% CO2 and 95% humidity) for 6 h and then, the O2/O3 treatment was repeated as described. After that, the cells plates were maintained in normoxia for 24 up to 72 h, before performing the experiments.
10
Hypoxia Assay for MDA-MB-231 Cells
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!