Measurement of peroxidase (POD) activity was performed by POD Activity Assay Kit (Solarbio, Beijing, China). POD activity was articulated as U/g. Measurement of superoxide dismutase (SOD) activity was performed by SOD Activity Assay Kit (Solarbio, Beijing, China). SOD activity was articulated as U/g. Measurement of catalase (CAT) activity was performed by CAT Activity Assay Kit (Solarbio, Beijing, China). CAT activity was articulated as U/g.
Cat activity assay kit
Manufactured by Solarbio
Sourced in China
The CAT activity assay kit is a laboratory tool used to measure the activity of the enzyme catalase (CAT) in biological samples. The kit provides the necessary reagents and protocols to quantify the catalase activity, which plays a crucial role in the antioxidant defense system of cells.
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Lab products found in correlation
Pod activity assay kit, by Solarbio (2 mentions)
Sod activity assay kit, by Solarbio (2 mentions)
Bca protein concentration assay kit, by Beyotime (1 mentions)
Mda content assay kit, by Solarbio (1 mentions)
H2o2 kit, by Solarbio (1 mentions)
Sod activity detection kit, by Solarbio (1 mentions)
Mda content detection kit, by Solarbio (1 mentions)
Spad 502 meter, by Konica Minolta (1 mentions)
5 protocols using cat activity assay kit
1
Enzymatic Activity Quantification
2
Catalase Activity Measurement in Zebrafish
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At 120 hpf, 10 larvae from the same treatment were collected and homogenized with the extracts in the assay kit. The homogenates were centrifuged at 8000× g for 10 min at 4 °C and supernatants gathered for the assays. Protein concentration and CAT activity were measured using the BCA Protein Concentration Assay Kit (Beyotime, Shanghai, China) and CAT activity assay kit (Solarbio, Beijing, China), respectively.
3
Antioxidant Activity and Reactive Oxygen Species Quantification in Tobacco
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CAT, SOD, and POD activities and MDA content were measured using a CAT activity assay kit, SOD activity assay kit, POD activity assay kit, and MDA content assay kit (Solarbio, China), respectively, according to the manufacturer’s instructions. The O2− and H2O2 contents in tobacco leaves were measured using the O2− assay kit and H2O2 kit (Solarbio, China), respectively, according to the manufacturer’s instructions. For each assay, three seedlings were separately measured.
Histochemical staining of O2− and H2O2 was performed using DAB and NBT, respectively, according to previously described procedures [65 (link)]. Briefly, discs of 1 cm diameter were cut from the largest leaf of each seedling, then stained with 50 mM sodium phosphate (pH 7.5) containing 0.2% NBT or 10 mM sodium phosphate (pH 6.5) containing 1 mg/mL DAB, then vacuumed for 10 min and incubated at 37 °C for 10 h under dark or light conditions, respectively. After that, the leaf discs were washed with ethanol, rinsed in boiling water for 10 min to remove chlorophyll, and then transferred into fresh ethanol. Finally, the stained leaves were photographed using a camera.
Histochemical staining of O2− and H2O2 was performed using DAB and NBT, respectively, according to previously described procedures [65 (link)]. Briefly, discs of 1 cm diameter were cut from the largest leaf of each seedling, then stained with 50 mM sodium phosphate (pH 7.5) containing 0.2% NBT or 10 mM sodium phosphate (pH 6.5) containing 1 mg/mL DAB, then vacuumed for 10 min and incubated at 37 °C for 10 h under dark or light conditions, respectively. After that, the leaf discs were washed with ethanol, rinsed in boiling water for 10 min to remove chlorophyll, and then transferred into fresh ethanol. Finally, the stained leaves were photographed using a camera.
4
Antioxidant Enzyme Activity Assays
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SOD activity was determined using a CheKine™ SOD Activity Assay Kit. Briefly, 0.1 g of alfalfa leaves was added to 1 mL of pre-cooled 1× lysis buffer and ground. After thorough grinding with a glass homogenizer, the sample was centrifuged for 5 min at 12,000 g at 4°C. The supernatant was collected as the sample to be tested. The procedure was performed according to the instruction manual, and the SOD content was calculated.
POD activity was determined using a CheKine™ POD Activity Assay Kit. The alfalfa leaves were washed with cold PBS. The water was removed, and the leaves were cut into pieces. A 0.1 g leaf sample was added to 1 mL of pre-cooled extraction buffer and centrifuged at 8000 g for 10 min at 4°C. The supernatant was collected for measurement. The procedure was performed following the manufacturer’s instructions, and the POD content was calculated.
CAT activity was determined using a Solarbio CAT Activity Assay Kit. The ratio of the mass of fresh alfalfa tissue (g) to the volume of the extraction solution (mL) was 1:5–10. The mixture was homogenized in an ice bath and centrifuged at 8000 g for 10 min at 4°C. The supernatant was collected and placed on ice to be measured. The CAT content was determined according to the manufacturer’s instructions and calculated.
POD activity was determined using a CheKine™ POD Activity Assay Kit. The alfalfa leaves were washed with cold PBS. The water was removed, and the leaves were cut into pieces. A 0.1 g leaf sample was added to 1 mL of pre-cooled extraction buffer and centrifuged at 8000 g for 10 min at 4°C. The supernatant was collected for measurement. The procedure was performed following the manufacturer’s instructions, and the POD content was calculated.
CAT activity was determined using a Solarbio CAT Activity Assay Kit. The ratio of the mass of fresh alfalfa tissue (g) to the volume of the extraction solution (mL) was 1:5–10. The mixture was homogenized in an ice bath and centrifuged at 8000 g for 10 min at 4°C. The supernatant was collected and placed on ice to be measured. The CAT content was determined according to the manufacturer’s instructions and calculated.
5
Physiological Responses in Plant Stress
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The leaf RWC was determined according to Galmés et al. [83 (link)]. The chlorophyll content was measured using a SPAD 502 meter (Konica-Minolta, Japan). The SOD activity was determined by a SOD activity detection kit (Solarbio, Beijing, China). The CAT activity was determined by a CAT activity assay kit (Solarbio, Beijing, China). The MDA content was determined using an MDA content detection kit (Solarbio, Beijing, China), and the content of Pro was quantified according to the instructions of a Pro content detection kit (Solarbio, Beijing, China). Compared with the control, the increase/decrease ratio of physiological indexes including the content of RWC, chlorophyll, and Pro, and the enzyme activity of SOD, CAT, and MDA were calculated using the formula: (experimental group − control group)/control group × 100% (n = 3).
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