Anaeropack
Manufactured by Thermo Fisher Scientific
Sourced in United States
The AnaeroPack is a laboratory equipment used to create an anaerobic environment. It is designed to maintain an oxygen-free atmosphere for the cultivation and incubation of anaerobic microorganisms.
Automatically generated - may contain errors
Lab products found in correlation
Buffered peptone water (bpw), by HiMedia (2 mentions)
Sheep blood, by Thermo Fisher Scientific (2 mentions)
Prism 9, by GraphPad (1 mentions)
Pm1 and pm2 plates, by Biolog (1 mentions)
Cc 3156, by Lonza (1 mentions)
Gentamicin, by Thermo Fisher Scientific (1 mentions)
Saponin, by Merck Group (1 mentions)
Deta nonoate, by Cayman Chemical (1 mentions)
9 protocols using anaeropack
1
Evaluating Candida albicans Filamentation
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C. albicans isolates were grown in YPD medium and then resuspended in sterile water to an OD600 of 0.2. The cell suspension was diluted 1:48 into inoculating fluid (IFY-0), and 100 μl of the cell suspension was aliquoted into each well of Biolog PM1 and PM2 plates according to the manufacturer’s instructions (Biolog Inc., Hayward, CA). The plate cultures were grown at 37°C for 24 h on a shaking platform at 200 rpm either aerobically or anaerobically (using Thermo Fisher AnaeroPack anaerobic gas generators in a sealed plastic bag). Following incubation, wells were scored for filamentation on a scale of 1 to 5 representing the proportion of filamentous cells in the population (1, 0 to 20%; 2, 20 to 40%; 3, 40 to 60%; 4, 60 to 80%; 5, 80 to 100%). PM experiments were performed with biological duplicates with growth (OD600) and filamentation scores averaged across the two replicates. Correlation analyses between growth and filamentation were performed using a simple linear regression model in GraphPad Prism 9.
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2
Quantifying Anaerobic Intestinal Bacteria
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On 7 dpi (d 21), one chicken per cage was euthanized, and approximately 20 g of jejunal contents (lower jejunal to upper ileal) were collected in sterile Whirl-Pak filter bags (Nasco, Fort Atkinson, WI). To each filter bag containing the jejunal content, 10 mL of buffered peptone water (BPW; Himedia, Mumbai, India) was added, and the samples were homogenized using a Masticator Silver Panoramic (Neutec Group Inc., Farmingdale, NY) for 1 min. Following homogenization, 1 mL of the mixed jejunal contents was transferred to a sterile glass dilution tube and diluted to 10−8 by serial dilution with BPW. Subsequently, 100 µL of the diluted contents (10−2, 10−4, 10−6, and 10−8) were plated on Tryptose Sulfite Cycloserine and Shahadi Ferguson Perfringens (TSC/SFP; Oxoid Ltd., Hampshire, United Kingdom) agar. The plates were incubated at 37°C for 24 h under anaerobic conditions created using an anaerobic gas-pack system (AnaeroPack™, Thermo Scientific, MA).
3
Anaerobic Cultivation and Metabolomics of F. rodentium
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F. rodentium anaerobic bacteria were acquired from the RIKEN BioResource Research Center (Tsukuba, Ibaraki, Japan) and inoculated in modified PYG mediumat 120 rpm for 1–2 days at 37 °C. The PYG medium was spun at 1500 g for 10 min, they were washed in 0.9% NaCl. Using an AnaeroPack (ThermoScientific, Massachusetts, USA), the reaction blend (0.1 M phosphate buffer (pH 7.2) and 0.2/0.4/1 mg/mL arachidonic acid) was thoroughly mixed with the obtained species in an anaerobic system. 50 μL of the centrifuged sample was mixed with equal proportion of cold ethyl acetate and isopropanol, then centrifuged once more for 5 min at 13,000 rpm. Targeted metabolomics-like derivatization techniques were employed for analysis.
4
Enumerating Intestinal C. perfringens
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Intestinal C. perfringens colony counting was conducted on two sampling days (6 and 8 dpi), following a previously reported study (Goo et al., 2023b ) with slight modifications. In brief, one chicken per cage was euthanized by cervical dislocation, and approximately 20 g of intestinal contents (from the middle of the jejunum to the end of the ileum) were collected using filter bags (While-Pak, Nasco, Fort Atkinson, WI). Next, 10 mL of 0.1% buffered peptone water (BPW; Himedia, Mumbai, India) was added to each filter bag and homogenized for 60 sec using a homogenizer (Masticator Silver Panoramic, Neutec Group Inc., Farmingdale, NY). The homogenized intestinal content was then transferred into a sterile dilution tube and diluted to 10−8 by a serial dilution. Subsequently, 100 μL of the diluted intestinal contents in each dilution tube (10−4, 10−6, and 10−8) were dispensed onto Tryptose Sulfite Cycloserine and Shahadi Ferguson Perfringens (TSC/SFP; Oxoid Ltd., Hampshire, UK) agar plates and gently spread. The TSC/SFP agar plates were then anaerobically incubated (AnaeroPack, Thermo Scientific, MA) at 37℃ for up to 48 h. After the incubation, the colonies were counted and recorded.
5
Culturing and Diluting P. gingivalis
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P. gingivalis ATCC 33277 was cultured in an anaerobic jar with an AnaeroPack (Thermo Fisher Scientific Inc.) generated anaerobic environment. Brain heart infusion (BHI) medium supplied with hemin and vitamin K was used to culture P. gingivalis. Three‐day‐old P. gingivalis at logarithmic growth phase were harvested by centrifugation at 5000 × g for 10 min. After resuspending with PBS, P. gingivalis was diluted with PBS to adjust the optical density to 0.8 at 660 nm, equalling to 109 CFU/ml. P. gingivalis was incubated with cells at different multiplicities of infection (MOI).
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P. gingivalis ATCC 33277 was cultured in an anaerobic jar with an AnaeroPack (Thermo Fisher Scientific Inc.) generated anaerobic environment. Brain heart infusion (BHI) medium supplied with hemin and vitamin K was used to culture P. gingivalis. Three‐day‐old P. gingivalis at logarithmic growth phase were harvested by centrifugation at 5000 × g for 10 min. After resuspending with PBS, P. gingivalis was diluted with PBS to adjust the optical density to 0.8 at 660 nm, equalling to 109 CFU/ml. P. gingivalis was incubated with cells at different multiplicities of infection (MOI).
6
Hypoxia-reoxygenation in brain endothelial cells
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Human brain microvascular endothelial cells were purchased from Applied Cell Biology Research Institute (Kirkland, WA, USA). OGD/R was performed as previously described [14 (link)]. Confluent cells were washed and replaced with glucose-free Dulbecco’s Modified Eagle’s Medium (DMEM). Cells were then incubated with AnaeroPack (AN0025A; Thermo Fisher Scientific) for 6 h. After OGD, cells were reperfused by placing them in endothelial basal medium 2 (CC-3156; Lonza, Walkersville, MD, USA) containing vehicle or CU06-1004 (10 μg/mL) and were incubated in a 95% air/5% CO2 for 16 h. After 16 h, immunofluorescence staining was performed.
7
Anaerobic Culture and Gavage of A. muciniphila
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A. muciniphila (ATCC BAA-835) was cultured in sterilized brain heart infusion broth (BD Difco) at 37°C in an airtight pot with AnaeroPack (Thermo Fisher Scientific) for approximately 48 h to reach a late exponential growth phase under strict anaerobic conditions. Cultures were centrifuged at 11,500 g for 10 min, washed with sterile PBS for twice, and then the bacterial cells were re-suspended with sterile PBS to 10 8 colony-forming units (CFUs)/200 μL. It was placed on ice immediately before administering to each mouse by gavage.
8
Quantifying Intracellular Bacterial Killing
9
Oxygen and nitrogen donor exposure on C. acnes
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