IL-1β and IL-18 levels in kidney were assessed using a rat ELISA kit (Elabscience Biotechnology Co., Ltd., Wuhan, China) according to the manufacturer's instructions. Caspase-1 activity in HK-2 cells was determined using an enzyme activity assay kit (Beyotime Biotechnology, Shanghai, China), according to the manufacturer's instructions.
Enzyme activity assay kit
Manufactured by Beyotime
Sourced in China
The Enzyme Activity Assay Kit is a laboratory instrument designed to measure the activity of enzymes. It provides a standardized procedure for quantifying the catalytic function of enzymes in a sample.
Automatically generated - may contain errors
3 protocols using enzyme activity assay kit
1
Measuring Inflammatory Mediators in Kidney
2
Probing ROS Generation in Fungal Hyphae
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The generation of ROS in hyphae was observed with the ROS-specific fluorescent probe 2, 7-dichlorodihydrofluorescein diacetate (DCFH-DA, Beyotime Biotechnology, Haimen, China) [35 (link)]. DCFH-DA at 10 µM was added to the mycelium cultures and then incubated in the dark for 30 min. The mycelium was washed three times with deionized water, and the fluorescence was observed under an Olympus fluorescent microscope (CKX41, Tokyo, Japan) with an excitation/emission wavelength of 485/528 nm. Hydrogen peroxide (H2O2) and superoxide anion (O2−) contents in mycelia were determined as previously described [36 (link),37 (link)]. The activities of superoxide dismutase (SOD), catalase (CAT) and NADPH oxidase (NOX) were measured using the Enzyme Activity Assay Kit (Beyotime Biotechnology, Nanjing, China) following the manufacturer’s instructions and previous reports [38 ,39 (link)]. The activity of peroxidase (POD) was determined as previously described [40 (link)]. To analyze the role of ROS in MLT treatment, exogenous H2O2 (100 μM) and vitamin C (Vc) (10 μM) were used as an ROS donor and scavenger, respectively, and their dosages were chosen based on our previous studies [30 (link)]. They were added to the culture to pre-treat for 30 min before the addition of MLT.
3
Oxidative Stress Response in Mycelia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The ROS-specific fluorescent probe 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA, Beyotime, Haimen, China) was added at 10 μM in the mycelia cultures in the darkness for 30 min, then the ROS accumulation in mycelia was detected under a CKX41 fluorescent microscope (Olympus, Tokyo, Japan) with an excitation/emission wavelength of 485/528 nm [25 ]. The content of hydrogen peroxide (H2O2) in mycelia was determined as previously described by Mirshekari et al. (2019) [26 ]. The activities of superoxide dismutase (SOD), NADPH oxidase (NOX) and catalase (CAT) were determined using the Enzyme Activity Assay Kit (Beyotime, Nanjing, China) according to the manufacturer's protocols and previous reports [27 ,28 ]. The activity of peroxidase (POD) was determined as previously described method by Wu et al. (2002) [29 (link)].
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!