AGS cells were cultured in two 12 well plates and incubated with lactobacilli (MOI = 100) and H. pylori P12-GFP (MOI = 100) for 6h. At the end of incubation, one plate of the cells was washed thrice with PBS and photographed by fluorescence and white light. A Urease detection reagent was prepared by adding concentrated hydrochloric acid into the PBS (pH=7.4) to adjust the solution to pH=6.8. Urea (Solarbio) and phenol red (Solarbio) were weighed and added to reach concentrations of 110mmol/L and 10mg/L, respectively, and then dissolved by vigorously shaking. To the other 12well plates, 1 ml Urease detection reagent was added. After 1-7 hours of reaction, 80μl medium was withdrawn to record the absorbance value at 540nm(Shmuely et al., 2004 (link); Rokka et al., 2008 (link); Tharmalingam et al., 2014 (link); Yang et al., 2020 (link)).
Phenol red
Manufactured by Solarbio
Sourced in China
Phenol red is a chemical compound commonly used as a pH indicator in various laboratory applications. It exhibits different colors at different pH levels, making it useful for monitoring and detecting changes in the acidity or alkalinity of solutions.
Automatically generated - may contain errors
Lab products found in correlation
Urea, by Solarbio (1 mentions)
Citric acid, by Beijing Chemical Works (1 mentions)
Malondialdehyde mda elisa kit, by Nanjing Jiancheng (1 mentions)
Pyruvate, by Solarbio (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Hank s solution, by Thermo Fisher Scientific (1 mentions)
Dmem media, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Solarbio (1 mentions)
4 protocols using phenol red
1
Quantifying Urease Activity in H. pylori
2
Antioxidant Biomarker Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following chemicals were purchased from the suppliers given in brackets: citric acid (Beijing Chemical Reagent Company, batch No.: 060403), ammonia (Sinopharm Chemical Reagent Co., Ltd., batch No.: 20120918), phenol red (Solarbio Company, batch No.: 20114.12.07), nutrient broth (China Pharmaceutical and Biological Products), ether (Beijing Reagent Company, batch No.: 20090707), superoxide dismutase (SOD) ELISA kit (Nanjing Jiancheng Institute of Biotechnology, batch No.: 201209), malondialdehyde (MDA) ELISA kit (Nanjing Jiancheng Institute of Biotechnology, batch No.: 201209), and micro BCA protein assay kit (Kangwei Century Company, batch No.: 201209).
3
Ovine Granulosa Cell Glucose Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
When cell confluence reached 80% medium, the original medium is removed. Cells were washed with 1 × PBS, and then, all treatments were cultured in DMEM, which was prepared with no serum, glucose, pyruvate, and phenol red (Solarbio, Beijing, China) for 8 h. Subsequently, the cells were supplemented with 8.4 mM and 33.6 mM of glucose cultured for an additional 24 h. The 8.4 mM represented an optimum glucose concentration for the secretion of steroid hormones by ovine GCs in vitro [6 (link),7 (link)], and the 33.6 mM group represents 30 times the physiological concentration of glucose in follicular fluid and was used to detect changes in steroid hormones at super-physiological concentrations [48 (link),49 (link),50 (link)]. This culture system was developed so that GC retains hormonally responsive aromatase activity and does not luteinize with time in culture [51 (link),52 (link),53 (link),54 (link)]. The GCs were collected for subsequent RNA-seq after the 24 h treatment period.
4
Cultivating Virus-Infected MDBK Cells for Transcriptomic Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The frozen MDBK cells (13 passages) were removed from liquid nitrogen and seeded onto T25 breathable cell culture flask (NEST, China) containing Dulbecco’s modified eagle medium with 10% Fetal bovine serum (Gibco, USA) at 37°C in 5% CO2 atmosphere for 24 to 48 h in the cell incubator. When the cell confluence reached 80 to 90%, the cells were digested with Trypsin-EDTA (0.25%) without phenol red (Solarbio, China), then passaged with the ratio of 1:2 in 10 cm cell culture dishes.
When the concentration of cells reached 1 × 107 cells per dish, 1 MOI of BoHV-1 was inoculated to MDBK cells and cultured in DMEM media (Gibco, USA) without serum at 37°C in 5% CO2 atmosphere. Cytopathic effects (CPE) were observed by light microscopy when virus-infected cells were cultured for 18, 24, and 33 h. The samples were harvested when the CPE reached varying degrees of monolayer at 18, 24 and 33 h. The sample from each dish was washed three times with Hank’s solution (Gibco, USA) and digested with 1 mL TRIzol reagent (Life, USA). The sample was then collected into nuclease-free cell cryopreservation tubes and sealed with All-Purpose Parafilm M Laboratory Film (Parafilm, USA) before being stored in liquid nitrogen for quick freezing. Finally, the samples were sequenced by Guangzhou Genedenovo Biotechnology Co., Ltd (Guangzhou, China).
When the concentration of cells reached 1 × 107 cells per dish, 1 MOI of BoHV-1 was inoculated to MDBK cells and cultured in DMEM media (Gibco, USA) without serum at 37°C in 5% CO2 atmosphere. Cytopathic effects (CPE) were observed by light microscopy when virus-infected cells were cultured for 18, 24, and 33 h. The samples were harvested when the CPE reached varying degrees of monolayer at 18, 24 and 33 h. The sample from each dish was washed three times with Hank’s solution (Gibco, USA) and digested with 1 mL TRIzol reagent (Life, USA). The sample was then collected into nuclease-free cell cryopreservation tubes and sealed with All-Purpose Parafilm M Laboratory Film (Parafilm, USA) before being stored in liquid nitrogen for quick freezing. Finally, the samples were sequenced by Guangzhou Genedenovo Biotechnology Co., Ltd (Guangzhou, China).
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!