ELISA assay was performed to evaluate the levels of brain oxidative damage and ferroptosis indexes [32 (link)]. After cardiac perfusion, the ipsilateral brain tissues were homogenized with cold lysis buffer, and the supernatant was separated via centrifugation at 4 °C at 14,000 rpm for 30 min. The levels of malondialdehyde (MDA) (S0131), antioxidant enzyme superoxide dismutase (SOD) (S0101), and glutathione (GSH)/oxidized glutathione (GSSG) (S0053) were measured using ELISA assay kits according to the manufacturer’s instructions (Beyotime Biotechnology, Shanghai, China). Briefly, MDA detection was based on the chromogenic reaction between MDA and thiobarbituric acid, and measured at 535 nm light wave [33 (link)]. SOD analysis was performed according to the WST-8 method and detected at 450 nm light wave [34 (link),35 (link)]. The GSH/GSSG assay was performed by glutathione reductase treatment and the concentration is detected at 412 nm light wave [33 (link)].
Elisa assay kit
Manufactured by Beyotime
Sourced in China
ELISA assay kits are laboratory equipment used to detect and quantify the presence of specific proteins or other analytes in a sample. The core function of these kits is to provide a standardized and reliable method for performing enzyme-linked immunosorbent assays (ELISA), a widely used analytical technique in various fields, including clinical diagnostics, research, and drug discovery.
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Lab products found in correlation
8 protocols using elisa assay kit
1
Oxidative Damage and Ferroptosis Assessment in Brain Tissue
2
Quantifying Inflammatory Cytokines in Spinal Cord
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The spinal cord tissues were minced firstly, and homogenized using lysis buffer. After centrifugation (30 min at 4° C), the supernatant samples were used to measure the expression of IL-6 (#PI328), IL-1β (#PI303), and TNF-α (#PT516) with related ELISA assay kits (Beyotime, China).
3
Evaluating Inflammatory Response in ICH
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To evaluate the degree of inflammation, enzyme-linked immunosorbent assay (ELISA) was performed to detect inflammatory factors (IL-1β, IL-6, and TNF-α) in all four groups. At 24 hours after ICH, the mice were anesthetized, and the brain tissue surrounding the hematoma was collected using an inverted microscope (RWD Life Science). ELISA assay kits (Beyotime, Shanghai, China, Cat# PI301, PI326, PT512) were used to detect the expression of inflammatory factors (IL-1β, IL-6, and TNF-α) according to the manufacturer‘s instructions. The concentration of each protein was calculated based on absorbance at 450 nm.
4
Evaluating Oxidative Stress in SH-SY5Y Cells
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SH-SY5Y cells were treated with PBS or 1 mM of MPP+ for 24 h, or transfected with si-NC or si-NEAT1 prior to MPP+ treatment. After 24 h treatment, reactive oxygen species (ROS) generation, lactate dehydrogenase (LDH) activity and superoxide dismutase (SOD) activity were measured using corresponding commercial ELISA assay kits (Beyotime, Shanghai, China) in accordance with the manufacturer's guidance.
5
Oxidative Stress and Inflammation in H/R-Induced Cardiomyocytes
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As specified in the kit instructions, all steps were followed strictly. Expression levels of oxidative stress factors (SOD and MDA), as well as pyroptosis-related proteins (GPX4 and SLC7A11), and inflammatory cytokines (TNF-α, IL-1β, IL-10 and IL-10) in H/R-induced HCMs and serum of MI/R mice, were detected by the respective ELISA assay kits (all purchased Beyotime).
6
Pramipexole's Effects on Neurodegeneration
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Pramipexole (HY-17355) was obtained from MedChemExpress (Monmouth Junction, NJ, USA). Nissl Staining Solution and ELISA Assay Kit were provided by Beyotime (Shanghai, China). The primary antibodies used in this study were anti-GADPH (ab9485; Abcam, Cambridge, UK), anti-DRD2 (DF10211; Affinity, Shanghai, China), anti-Bax (ab32503; Abcam, Cambridge, UK), anti-NeuN (ab196495; Abcam, Cambridge, UK), anti-Caspase-3 (BS1518; Bioworld, Nanjing, China) and anti-Bcl-2 (ab196495; Abcam, Cambridge, UK). Goat anti-rat IgG (HRP) (SA00001-2; Proteintech, Wuhan Shi, China) was used as the secondary antibody for western blotting.
7
Glucose Metabolism Assays in CRC Cells
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RKO and HCT116 cells were transfected with shRNA expression vectors or overexpression vectors. Glucose level in CRC cells was determined with a Glucose content assay kit. Lactate production was measured with a lactate assay kit.
G6PD activity was assessed with a
G6PD assay kit. The assay kits mentioned above were purchased from Cominbio (Suzhou, China). Intracellular NADPH level was assessed with an NADPH content assay kit (Solarbio, Beijing, China). To assess
G6PD activity in CRC samples, tissues were homogenized, and enzyme activity was assessed with the
G6PD assay kit and calculated relative to that of the control group. The total protein amount was measured using the ELISA assay kit (Beyotime, Shanghai, China).
G6PD activity was assessed with a
G6PD assay kit. The assay kits mentioned above were purchased from Cominbio (Suzhou, China). Intracellular NADPH level was assessed with an NADPH content assay kit (Solarbio, Beijing, China). To assess
G6PD activity in CRC samples, tissues were homogenized, and enzyme activity was assessed with the
G6PD assay kit and calculated relative to that of the control group. The total protein amount was measured using the ELISA assay kit (Beyotime, Shanghai, China).
8
Quantifying Inflammatory Cytokines in Mouse Hypothalamus
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ELISA assay kit purchased from Beyotime Institute of Biotechnology was used according to the manufacturer’s instructions. The mice were sacrificed by decollation. The hypothalamus was homogenized in PBS (pH 7.4) and then centrifuged. The supernatant was recovered and used as the test sample. Monoclonal capture antibodies specific to mouse TNF-α, IL-6, and IL-1β have been precoated on the plate, and mouse TNF-α, IL-6, and IL-1β binds to the capture antibody when added to the standard or sample. Then after a series of steps according to the manufacturer’s instructions, the absorbance of each well was recorder at 450 nm.
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