A084 3 1

Manufactured by Nanjing Jiancheng

Sourced in China

The A084-3-1 is a laboratory equipment product. It serves as a core function, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

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A084-3-1 Peroxidase assay kit A084-3

10 protocols using a084 3 1

Physiological Profiling of Diploid and Tetraploid Leaves

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The fully expanded mature leaves of diploids and tetraploids were selected for physiological parameter measurements. For the determination of the catalase (CAT) activity, peroxidase (POD) activity, malondialdehyde (MDA) content, total protein content, and soluble sugar content, their corresponding assay kits (Catalog No. A007-1-1, A084-3-1, A003-1-2, A045-2-2, and A145-1-1, Nanjing Jiancheng Bioengineering Ins., Nanjing, China) were used according to the manufacturer’s protocols.

Liu Z., Wang J., Qiu B., Ma Z., Lu T., Kang X, & Yang J. (2022). Induction and Characterization of Tetraploid Through Zygotic Chromosome Doubling in Eucalyptus urophylla. Frontiers in Plant Science, 13, 870698.

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Salt Stress Impacts Antioxidant Enzymes

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The seeds from WT, VC and PePLDδ-overexpressed lines (OE6 and OE7) were germinated and cultured on 1/2 MS medium for seven days. These seedlings were then treated with 0 or 130 mM NaCl for another seven days. Control and salt-stressed seedlings were harvested and ground in liquid-nitrogen-precooled mortars. The samples (0.1 g fresh weight) were mixed with 1 mL precooled extraction buffer containing 1mM EDTA, 1% PVP, 1mM ASA and 50 mM potassium phosphate buffer (pH 7.0). Through centrifugation (12,000 g) at 4 °C for 10 min, the supernatant solution was obtained to determine enzyme activities of superoxide dismutase (SOD), peroxidase (POD) and ascorbic acid peroxidase (APX). Antioxidant enzyme activity assay kits, such as A001-3-2 (total SOD determination kit), A084-3-1 (POD assay kit) and A123-1-1 (APX test box) (Nanjing Jiancheng Bioengineering Institute, Nanjing, China), were used to detect enzymatic activity according to the manufacturer instructions. The total protein in crude enzyme extract was assayed with A045-2-2 (total protein determination kit) (Nanjing Jiancheng Bioengineering Institute, Nanjing, China).

Zhang Y., Yao J., Yin K., Liu Z., Zhang Y., Deng C., Liu J., Zhang Y., Hou S., Zhang H., Yu D., Zhao N., Zhao R, & Chen S. (2022). Populus euphratica Phospholipase Dδ Increases Salt Tolerance by Regulating K+/Na+ and ROS Homeostasis in Arabidopsis. International Journal of Molecular Sciences, 23(9), 4911.

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Antioxidant Enzyme Activities in Potato Leaves

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The activity of PAL was used as an important metabolic process indicator in potatoes. The total SOD activity in potato leaves was determined using a kit (A001-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China) based on the nitrobluetetrazolium method according to the manufacturer’s instruction, as shown in a previous report [37 (link)]. The POD activity was measured by using a POD assay kit (A084-3-1; Nanjing Jiancheng Bioengineering Institute, Najing, China) on the basis of guaiacol oxidation at 470 nm by H₂O₂ according to the manufacturer’s instruction, as shown in a previous report [38 (link)]. Similarly, the activity of CAT was also measured using a CAT assay kit (A007-1; Nanjing Jiancheng Bioengineering Institute, China) as seen in a report [38 (link)]. The PAL can catalyze L-phenylalanine to produce trans-cinnamic acid and ammonia, and the maximum absorption peak of trans-cinnamic acid is 290 nm. PAL activity was calculated by measuring the increase in the OD value to 290 nm using a PAL assay kit (E-BC-K522-S; Elabscience Biotechnology Co. Ltd, China). As there was no POD, CAT, and PAL activity in P. infestans through test kits, the SOD activity of P. infestans was used as the antioxidant system indicator in P. infestans.

Wang Z.L., Zhang X., Fan G.J., Que Y., Xue F, & Liu Y.H. (2022). Toxicity Effects and Mechanisms of MgO Nanoparticles on the Oomycete Pathogen Phytophthora infestans and Its Host Solanum tuberosum. Toxics, 10(10), 553.

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Maize Leaf Antioxidant Enzyme Analysis

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Fresh leaves (the ear leaf) of maize were taken at the tasseling stage for the determination of leaf antioxidant enzyme activities and malondialdehyde (MDA) content. Specifically, 0.2 g of fresh leaves were placed in a precooled mortar, and 1.8 mL phosphate buffer (pH = 7.8) was added to the ice bath to grind the leaves into a homogenate. The grinding solution was transferred into a 2 mL centrifuge tube and centrifuged at 12000 r·min^-1 for 10 min at 4°C. The supernatant, which was the crude enzyme extract, was stored at -20°C for further use for the determination of peroxidase (POD) and superoxide dismutase (SOD) activities. Alternatively, the phosphate buffer used above was replaced with saline extract to obtain the enzyme extract for the determination of catalase (CAT) activity. The extract was replaced with the extract from the MDA kit, and the obtained enzyme extract was used for the determination of MDA content.
Then, the enzyme extracts were further processed for POD, SOD, CAT and MDA measurements using A084-3-1, T001-1, A007-1-1 and A003-3-1 kits (Nanjing Jiancheng Bioengineering Institute), respectively. Finally, the absorbance values were measured at 420 nm, 550 nm, 405 nm and 532 nm using a plate reader (Synergy HTX, BioTek, USA).

Guo Z., Ye W., Wang H., He W., Tian Y., Hu G., Lou Y., Pan H., Yang Q, & Zhuge Y. (2024). Straw and phosphorus applications promote maize (Zea mays L.) growth in saline soil through changing soil carbon and phosphorus fractions. Frontiers in Plant Science, 15, 1336300.

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Physiological Response Assays for Salt-Stressed Plants

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The physiological indicators: PAL, H₂O₂, MDA, Pro, and POD were measured using detection assay kits (Cat. No. A137-1-1, A064-1-1, A003-3-1, A107-1-1 and A084-3-1 respectively; Nanjing Jiancheng Bioengineering Institute, China). All the tests were performed according to the manufacturer’s instructions (http://www.njjcbio.com/, accessed on 1 December 2022). A lignin test kit was obtained from COMIN company (Cat. No. MZS-1-G; Suzhou, China). The leaves were used for physiological detection. Expanded leaves were collected from L96 and NT plants after salt stress (0 d, 2 d and 5 d). Each sample was repeated 4 times. At the same time, root were collected, washed, and frozen in −80 °C for the following RNA extraction and plant hormone detection via HPLC (Agilent 1260, Santa Clara, CA, USA).

Yang H., Yang Q., Zhang D., Wang J., Cao T., Bozorov T.A., Cheng L, & Zhang D. (2023). Transcriptome Reveals the Molecular Mechanism of the ScALDH21 Gene from the Desert Moss Syntrichia caninervis Conferring Resistance to Salt Stress in Cotton. International Journal of Molecular Sciences, 24(6), 5822.

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Evaluating Plant Response to Stress

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After the treatment period (63 days), the whole plant was uprooted, washed, and the fresh weight, root weight, root length, and plant height were measured. A root scanner (WinRHIZO STD4800 LA2400, Regent, Canada) was used to determine the root surface area. Forage leaves and soil samples were collected to determine enzyme activity. Leaf catalase, peroxidase, superoxide dismutase, malondialdehyde, H₂O₂, proline, and chlorophyll contents were determined using the corresponding kits (A007-1-1, A084-3-1, A001-1-2, A003-1-2, A064-1-1, A107-1-1, and A147-1-1, Nanjing Jiancheng Bio-Engineering Institute Co., Ltd., China). The activity of soil alkaline protease (ALPT), urease (UE), neutral phosphatase (NP), acid phosphatase (ACP), polyphenol oxidase (PPO), and sucrase (SC) were determined using soil enzyme activity kits (BC0885, BC0125, BC0465, BC0145, BC0115, and BC0245, Beijing Solarbio Science & Technology Co., Ltd., China).

Tang X., Fei X., Sun Y., Shao H., Zhu J., He X., Wang X., Yong B, & Tao X. (2022). Abscisic acid-polyacrylamide (ABA-PAM) treatment enhances forage grass growth and soil microbial diversity under drought stress. Frontiers in Plant Science, 13, 973665.

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Physiological Responses of Banana Seedlings

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Ion leakage was tested following the method reported by Jiang and Zhang [66 (link)]. The content of malondialdehyde (MDA) was examined based on the thiobarbituric acid colorimetric method [67 (link)]. Proline content was examined following Bates et al. [68 ]. The content of H₂O₂ was measured with a detection kit (W-100, G-CLONE, Beijing, China). The activities of perox
idase (POD), catalase (CAT) , and superoxide dismutase (SOD) were spectrophotometrically tested
with detection kits (A007-1-1, A084-3-1, and A001-1-1; Jiancheng, Nanjing, China).. The soluble sugar content was examined using a detection kit (A145-1-1, Jiancheng, Nanjing, China). The content of jasmonate and the activities of 1-aminocyclopropane-1-carboxylic acid synthase (ACS) and 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) were measured with detection kits (ml062281, ml062340-2, ml062341-2, Meilian, Shanghai, China). Banana seedlings were enclosed in an airtight container for 8 hours to collect ethylene. Ethylene production was examined by gas chromatography (GC-2010 Plus) following the manufacturer’s instructions.

Xu Y., Hu W., Song S., Ye X., Ding Z., Liu J., Wang Z., Li J., Hou X., Xu B, & Jin Z. (2022). MaDREB1F confers cold and drought stress resistance through common regulation of hormone synthesis and protectant metabolite contents in banana. Horticulture Research, 10(2), uhac275.

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Antioxidative Biomarker Determination in Plant Tissues

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A certain weight of leaf or shoot sample was crushed in a prechilled pestle and crushed with a mortar in an appropriate volume of prechilled extraction buffer. After uniform homogenization and centrifugation at 4000 g for 15 min at 4 °C, the supernatant was collected and used for biochemical analyses.
Antioxidative capabilities were estimated by quantifying peroxidation products and antioxidase activity levels. The malondialdehyde (MDA) content was determined using a Plant MDA Assay Kit (TBA method; A003-3-1, Nanjing Jiancheng, Nanjing, China). The hydrogen peroxide (H 2 O 2 ) level was determined using an H 2 O 2 Assay Kit (H 2 O 2 reacted with titanium salt and is then quantified at OD 415 ; A064-1-1, Nanjing Jiancheng), and the superoxide anion (O 2 -) level was determined using an O 2 -Assay Kit (sulfanilamide method; R30342, Shanghai Yuanye, Shanghai, China). The activity levels of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were determined using the Total SOD (WST-1 method; A001-1-2, Nanjing Jiancheng), POD (H 2 O 2 catalysis method; A084-3-1, Nanjing Jiancheng) and CAT (H 2 O 2 catalysis method; A007-1-1, Nanjing Jiancheng) Assay Kits, respectively, following the manufacturer's instructions. All the leaf and shoot samples were measured three times in parallel.

Sun H., Wu S., & Wu L. (2021). White oak (Quercus fabri Hance) regenerated stump sprouts show few senescence symptoms during 40 years of growth in a natural forest. Forest Ecosystems, 8(1).

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Antioxidant Capacity Analysis of Plant Extracts

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A certain weight of leaf or shoot sample was crushed in a prechilled pestle and crushed with a mortar in an appropriate volume of prechilled extraction buffer. After uniform homogenization and centrifugation at 4,000 g for 15 min at 4°C, the supernatant was collected and used for biochemical analyses.
Antioxidative capabilities were estimated by quantifying peroxidation products and antioxidase activity levels.
The malondialdehyde (MDA) content was determined using a Plant MDA Assay Kit (TBA method; A003-3-1, Nanjing Jiancheng, Nanjing, China). The hydrogen peroxide (H 2 O 2 ) level was determined using an H 2 O 2 Assay Kit (H 2 O 2 reacted with titanium salt and is then quanti ed at OD 415 ; A064-1-1, Nanjing Jiancheng), and the superoxide anion (O 2 -) level was determined using an O 2 -Assay Kit (sulfanilamide method; R30342, Shanghai Yuanye, Shanghai, China). The activity levels of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were determined using the Total SOD (WST-1 method; A001-1-2, Nanjing Jiancheng), POD (H 2 O 2 catalysis method; A084-3-1, Nanjing Jiancheng) and CAT (H 2 O 2 catalysis method; A007-1-1, Nanjing Jiancheng) Assay Kits, respectively, following the manufacturer's instructions. All the leaf and shoot samples were measured three times in parallel.

Sun H., Wu S., & Wu L. (2020). White Oak (Quercus fabri Hance) regenerated stump sprouts show few symptom of senescence during 40 years’ growth in the natural forest.

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Measuring Antioxidant Capacity in Plant Tissues

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A certain weight of leaf or shoot sample was crushed in a prechilled pestle and crushed with a mortar in an appropriate volume of prechilled extraction buffer. After uniform homogenization and centrifugation at 4,000 g for 15 min at 4°C, the supernatant was collected and used for biochemical analyses.
Antioxidative capabilities were estimated by quantifying peroxidation products and antioxidase activity levels. The malondialdehyde (MDA) content was determined using a Plant MDA Assay Kit (TBA method; A003-3-1, Nanjing Jiancheng, Nanjing, China). The hydrogen peroxide (H 2 O 2 ) level was determined using an H 2 O 2 Assay Kit (H 2 O 2 reacted with titanium salt and is then quantified at OD 415 ; A064-1-1, Nanjing Jiancheng), and the superoxide anion (O 2 -) level was determined using an O 2 -Assay Kit (sulfanilamide method; R30342, Shanghai Yuanye, Shanghai, China). The activity levels of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were determined using the Total SOD (WST-1 method; A001-1-2, Nanjing Jiancheng), POD (H 2 O 2 catalysis method; A084-3-1, Nanjing Jiancheng) and CAT (H 2 O 2 catalysis method; A007-1-1, Nanjing Jiancheng) Assay Kits, respectively, following the manufacturer's instructions. All the leaf and shoot samples were measured three times in parallel.

Sun H., Wu S., & Wu L. (2021). White oak (Quercus fabri Hance) regenerated stump sprouts show few senescence symptoms during 40 years’ growth in a natural forest.

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