human keap1 enzyme linked immunosorbent assay elisa kit  (Wuhan Fine Biotech)

Journal: Bioactive Materials

Article Title: A multimodal ROS logic-gated therapeutic platform disrupts the vicious cycle of senescence to promote aged bone defect repair

doi: 10.1016/j.bioactmat.2026.02.002

Figure Lengend Snippet: Transcriptomic and molecular analysis of the potential pathways involved in MMBOx-mediated BMSCs rejuvenation. (A) Circular heatmap showing differentially expressed genes (DEGs) associated with cell senescence, inflammation, and osteogenesis in senescent BMSCs treated with MMBOx@GPP compared to GPP. (B) Gene Ontology (GO) enrichment analysis of upregulated DEGs. (C) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of upregulated DEGs. (D) Gene Set Enrichment Analysis (GSEA) plots of the glutathione metabolic process (ES: enrichment score; NES: normalized enrichment score; FDR: false discovery rate). (E) Heatmap of DEGs enriched in aging-related GO terms. (F) Western blot analysis of Keap1, Nrf2, Nqo1, Gclc, and GAPDH protein expression in BMSCs. (G) Quantitative analysis of protein band intensities ( n = 3). (H) Representative flow cytometry plots of ThiolTracker™ fluorescence staining indicating intracellular glutathione levels. (I) Quantification of intracellular GSH/GSSG ratio in BMSCs ( n = 3). (J) Schematic diagram illustrating the proposed mechanism by which MMBOx attenuates BMSCs senescence via Nrf2 pathway activation and glutathione metabolism enhancement. Data are expressed as mean ± SD; ∗ P < 0.05, ∗∗∗ P < 0.001.

Article Snippet: Western blotting was employed to evaluate protein expression of key targets, including Keap1 (Affinity, AF5266; 1:1000), Nrf2 (Proteintech, 16396-1-AP; 1:1000), Nqo1 (Abcam, ab80588; 1:10,000), Gclc (Proteintech, 12601-1-AP; 1:25000) and GAPDH (Proteintech, 60004-1-Ig; 1:50,000).

Techniques: Western Blot, Expressing, Flow Cytometry, Fluorescence, Staining, Activation Assay

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: Deletion of microglial Sirt6 inhibited the NRF2-HO1 signaling and worsened the peroxidation damage. (A) Gene Ontology (GO) analysis was performed on RNA-Seq data from microglia sorted from Sirt6 MCKO and Sirt6 fl/fl control mice. (B) TAC, MDA, SOD, and GSH/GSSG levels at 5 days after LPS injection. n = 4 mice. (C) Gene Set Enrichment Analysis (GSEA) of RNA-Seq data profiled from microglia sorted from Sirt6 MCKO and Sirt6 fl/fl control mice. (D-E) Analysis of NRF2-HO1 and associated signaling proteins in sorted microglia. Protein levels of NRF2, KEAP1, HO-1, NQO1, NLRP3, Cleaved Caspase-3, and Cleaved IL-1β were assessed by Western blot (D) and quantified (E). n = 4 mice. Data are mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: RNA Sequencing, Control, Injection, Western Blot, Two Tailed Test

Journal: Frontiers in Physiology

Article Title: The possible mechanisms linking chronic obstructive pulmonary disease and coronary atherosclerosis based on coronary computed tomography angiography and animal experiments

doi: 10.3389/fphys.2026.1688832

Figure Lengend Snippet: Oxidative stress response and the KEAP1–NRF2 pathway levels are linked to CAS in COPD patients. (A) ELISA detection of KEAP1 and NRF2 levels in the serum of patients. (B) KEAP1, NRF2, NQO1, and HO-1 levels in the serum of patients determined via RT-qPCR. (C) Levels of ROS and MDA, and activities of SOD and catalase in the serum of patients determined using kits. The CAS group was used as a normalization control. Data among multiple groups were compared using one-way ANOVA, followed by Tukey’s post-hoc tests with corrections for multiple comparisons. CAS, coronary atherosclerosis; COPD, chronic obstructive pulmonary disease; ROS, reactive oxygen species; MDA, malondialdehyde; SOD, superoxide dismutase.

Article Snippet: Serum KEAP1 and Nrf2 levels were measured using a human KEAP1 enzyme-linked immunosorbent assay (ELISA) kit (EH4240, Wuhan Fine Biotech Co., Ltd., Wuhan, Hubei, China) and a human Nrf2 ELISA kit (abs551899, Absin Bioscience Inc., Shanghai, China) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Control

Journal: Frontiers in Physiology

Article Title: The possible mechanisms linking chronic obstructive pulmonary disease and coronary atherosclerosis based on coronary computed tomography angiography and animal experiments

doi: 10.3389/fphys.2026.1688832

Figure Lengend Snippet: KEAP1–NRF2-mediated oxidative stress participates in the occurrence of COPD combined with CAS in mice. (A) PFT for FRC, RI, Cdyn, and MV in mice. (B) The histopathological changes of lung tissues observed using H&E staining. (C) Serum lipid levels in mice measured via ELISA. (D) KEAP1, NRF2, NQO1, and HO-1 levels in the serum of mice determined via RT-qPCR. (E) Levels of ROS and MDA, and activities of SOD and catalase in the lung tissue of mice measured using kits. (F) Representative Western blotting of KEAP1, NRF2, NQO1, and HO-1 proteins in the lung tissue of mice and the quantification data. n = 6 mice for each treatment. Data among multiple groups were compared using one-way ANOVA, followed by Tukey’s post-hoc tests for multiple comparisons. COPD, chronic obstructive pulmonary disease; CAS, coronary atherosclerosis; PFT, pulmonary function test; FRC, functional residual capacity; RI, resistance index; Cdyn, dynamic compliance; MV, minute ventilation; ROS, reactive oxygen species; MDA, malondialdehyde; SOD, superoxide dismutase.

Article Snippet: Serum KEAP1 and Nrf2 levels were measured using a human KEAP1 enzyme-linked immunosorbent assay (ELISA) kit (EH4240, Wuhan Fine Biotech Co., Ltd., Wuhan, Hubei, China) and a human Nrf2 ELISA kit (abs551899, Absin Bioscience Inc., Shanghai, China) according to the manufacturer’s instructions.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Functional Assay