rabbit monoclonal antibodies against nrf2  (Proteintech)

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: 8-OHdG and Nrf2 Protein are Expressed Consistently in Various T Stages of Invasive Breast Carcinoma

doi: 10.31557/APJCP.2025.26.1.301

Figure Lengend Snippet: Nrf2 Staining was Observed in Invasive Breast Carcinoma of No Special Type (IBC-NST) Tumor Cell’s Nuclei (black arrow). (A) Negative staining. (B) Weakly positive staining. (C) Moderately positive staining. (D) Strongly positive staining. All figures were captured at 400x magnification. Scale bar: 50 µm.

Article Snippet: The primary antibodies were mouse monoclonal antibody against 8-OHdG (e-8) sc-393871 (Santa Cruz Biotechnology) at 1:100 dilution and rabbit monoclonal antibody IgG against Nrf2 bsm-52179R (Bioss Antibodies) at 1:100 dilution.

Techniques: Staining, Negative Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Formononetin promotes porcine oocytes maturation and improves embryonic development by reducing oxidative stress

doi: 10.3389/fcell.2024.1520429

Figure Lengend Snippet: Information about the primer sequences.

Article Snippet: And incubation with horseradish peroxidase-conjugated secondary antibodies at RT for 1 h. Primary antibodies were rabbit monoclonal antibodies against NRF2 (1:2000, #16396-1-AP, Proteintech), KEAP1 (1:2000, #10503-2-AP, Proteintech), and β-Actin (1:5000, #20536-1-AP, Proteintech).

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: Formononetin promotes porcine oocytes maturation and improves embryonic development by reducing oxidative stress

doi: 10.3389/fcell.2024.1520429

Figure Lengend Snippet: Effect of FMN on mitochondrial function in oocytes and early embryos. (A) The mitochondria distribution at MII stage were stained with Mito-Tracker Red; scale bar, 50 μm. (B) The relative abundance of mitochondria in oocyte were analyzed for control and FMN-treated group. (C) Relative mRNA levels of ATP5B, NRF2 and KEAP1 in oocytes. (D) 4-cell stage embryos stained with JC-1. Scale bar, 100 μm. (E) Relative levels of JC-1 Red/Green fluorescence intensity in embryos were analyzed for control and FMN-treated group. (F) Relative mRNA levels of TFAM and NRF1 in embryos. (G) ATP levels of 4-cell stage embryos in both control and FMN-treated groups. Data are presented as the mean ± standard deviation (SD). * p < 0.05,** p < 0.01, *** p < 0.001 vs. 0 μM FMN group.

Article Snippet: And incubation with horseradish peroxidase-conjugated secondary antibodies at RT for 1 h. Primary antibodies were rabbit monoclonal antibodies against NRF2 (1:2000, #16396-1-AP, Proteintech), KEAP1 (1:2000, #10503-2-AP, Proteintech), and β-Actin (1:5000, #20536-1-AP, Proteintech).

Techniques: Staining, Control, Fluorescence, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: Formononetin promotes porcine oocytes maturation and improves embryonic development by reducing oxidative stress

doi: 10.3389/fcell.2024.1520429

Figure Lengend Snippet: Effect of FMN on the expression levels of the Nrf2/Keap1 signaling pathway-related protein and genes in porcine blastocysts. (A) Porcine blastocysts incubated with or without FMN were stained with NRF2 (green) and DAPI (blue), scale bar, 50 μm. (B) Relative levels of NRF2 fluorescence intensity in blastocysts were analyzed for the control and FMN-treated group. (C) Relative mRNA expression levels of genes related to Nrf2/Keap1 pathway, NRF2, KEAP1, NQO1, UCHL1 in blastocysts. (D) Western blot analysis of NRF2 and KEAP1 protein expressions in blastocysts in the control and FMN-treated groups. Data were presented as the mean ± standard deviation (SD). ** p < 0.01,*** p < 0.001 vs. 0 μM FMN group.

Article Snippet: And incubation with horseradish peroxidase-conjugated secondary antibodies at RT for 1 h. Primary antibodies were rabbit monoclonal antibodies against NRF2 (1:2000, #16396-1-AP, Proteintech), KEAP1 (1:2000, #10503-2-AP, Proteintech), and β-Actin (1:5000, #20536-1-AP, Proteintech).

Techniques: Expressing, Incubation, Staining, Fluorescence, Control, Western Blot, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: Formononetin promotes porcine oocytes maturation and improves embryonic development by reducing oxidative stress

doi: 10.3389/fcell.2024.1520429

Figure Lengend Snippet: Summary of the effect of Formononetin (FMN) on porcine oocyte in vitro maturation and early embryo in vitro . This schematic view illustrates that during the IVM stage, FMN enhances oocyte maturation in vitro by up-regulating genes related to CCs expansion and antioxidants, reducing oxidative stress, and improving mitochondrial function. During the IVC stage, FMN promotes the nuclear expression of Nrf2 through the P62-Keap1-Nrf2 pathway, which exerts antioxidant function and enhances mitochondrial function. Then, FMN promotes cell proliferation, reduces apoptosis, and lowers autophagy levels.

Article Snippet: And incubation with horseradish peroxidase-conjugated secondary antibodies at RT for 1 h. Primary antibodies were rabbit monoclonal antibodies against NRF2 (1:2000, #16396-1-AP, Proteintech), KEAP1 (1:2000, #10503-2-AP, Proteintech), and β-Actin (1:5000, #20536-1-AP, Proteintech).

Techniques: In Vitro, Expressing

Journal: Antioxidants

Article Title: The Cerebral Protective Effect of Novel Erinacines from Hericium erinaceus Mycelium on In Vivo Mild Traumatic Brain Injury Animal Model and Primary Mixed Glial Cells via Nrf2-Dependent Pathways

doi: 10.3390/antiox13030371

Figure Lengend Snippet: Effect of Hericium erinaceus mycelium (HEM) and its isolated erinacine C on brain histological NF-E2-related factor 2 (Nrf2) protein expression in the mild traumatic brain injury (mTBI) animal model. Rats with mTBI were treated without or with oral HEM administration (108.5, 217 mg/kg) and intraperitoneal erinacine C injection (2 mg/kg). Rats were sacrificed, and the brains were separated. The results show representative brain sections stained for the control group (CL), rats with mTBI, HEM, and its isolated erinacine C treatment staining of the brain tissue. Evaluations of cytosol protein expression were quantitative in the cortex and subcortex zones. The positive stained areas from three randomly selected observation fields were evaluated. Levels of proteins were quantitatively estimated through immunohistochemical analysis by average integrated optical density. Data are expressed as the mean ± standard deviation of independent experiments. n = 6. * p < 0.05, as compared to the control group; # p < 0.05, as compared to the mTBI-treated group.

Article Snippet: Mouse monoclonal antibodies against NeuN, iba1, β-actin, catalase, thioredoxin, superoxide dismutase, and BDNF, and rabbit poly monoclonal antibodies against Nrf2 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Isolation, Expressing, Animal Model, Injection, Staining, Control, Immunohistochemical staining, Standard Deviation

Journal: Antioxidants

Article Title: The Cerebral Protective Effect of Novel Erinacines from Hericium erinaceus Mycelium on In Vivo Mild Traumatic Brain Injury Animal Model and Primary Mixed Glial Cells via Nrf2-Dependent Pathways

doi: 10.3390/antiox13030371

Figure Lengend Snippet: Chromatin immunoprecipitation-quantitative polymerase chain reaction (ChIP-qPCR) determination of binding of NF-E2-related factor 2 (Nrf2) to proximal promoters of genes altered in lipopolysaccharide (LPS)-induced mixed glia cells with erinacine C treatment. Erinacine C facilitated the binding of Nrf2 to the promoter regions of BDNF , CAT , TrxR , and SOD genes. The chromatin immunoprecipitation assay was performed using antibodies against Nrf2 and BDNF , CAT , TrxR , and SOD genes promoters (the target sites, as described in the Materials and Methods) in the precipitated DNA, which was amplified by qPCR using specific primer sets. BV2 cells were incubated with or without LPS and treated with erinacine C at various concentrations for 24 h. The effect of erinacine C treatment on specific genes was calculated by the ΔΔCt method. The quantitative data are presented as the mean ± standard deviation of three independent experiments in triplicate technical repeats. * p < 0.05, as compared to the control group. # p < 0.05, as compared to the LPS–treated group.

Article Snippet: Mouse monoclonal antibodies against NeuN, iba1, β-actin, catalase, thioredoxin, superoxide dismutase, and BDNF, and rabbit poly monoclonal antibodies against Nrf2 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, ChIP-qPCR, Binding Assay, Amplification, Incubation, Standard Deviation, Control

Journal: Antioxidants

Article Title: The Cerebral Protective Effect of Novel Erinacines from Hericium erinaceus Mycelium on In Vivo Mild Traumatic Brain Injury Animal Model and Primary Mixed Glial Cells via Nrf2-Dependent Pathways

doi: 10.3390/antiox13030371

Figure Lengend Snippet: Schematic presentation of the molecular mechanism of the action of Hericium erinaceus mycelium (HEM) and erinacine C on the protection against neuronal injury and microglia activation in rat models of mild traumatic brain injury (mTBI) through the Nrf2 pathway. In the nucleus, Nrf2 promotes transcriptional activation of antioxidants enzymes (brain-derived neurotrophic factor ( BDNF ), catalase ( CAT ), thioredoxin reductase ( TrxR ), and superoxide dismutase ( SOD ) by erinacine C by binding to the promoter regions of the downstream target genes, thereby triggering the antioxidant defense system and suppressing microglial activation in primary mixed-glia cultures. HEM and its components erinacine C in the treatment of mild brain traumatic injury (mTBI)-induced neuronal damage and microglia activation that could protect from neuronal injury.

Article Snippet: Mouse monoclonal antibodies against NeuN, iba1, β-actin, catalase, thioredoxin, superoxide dismutase, and BDNF, and rabbit poly monoclonal antibodies against Nrf2 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Activation Assay, Derivative Assay, Binding Assay

Journal: Pharmaceutical biology

Article Title: Antioxidative effect of Periplaneta americana extract on dextran sulfate sodium-induced ulcerative colitis through activation of the Nrf2 signal.

doi: 10.1080/13880209.2023.2220351

Figure Lengend Snippet: Figure 7. PAE activated the Nrf2 signaling pathway in DSS-induced UC colons. (A) Expression bands of nuclear Nrf2, cytoplasmic Nrf2, total Nrf2, GCLM, GPX2, HO-1, and NQO1 by western blotting. Expression levels of nuclear Nrf2 (B), cytosolic Nrf2 (C), GCLM (D), GPX2 (E), HO-1 (F), and NQO1 (G). Error bars represent the mean ± SEMs (n ¼ 3/group). ##p < 0.01, #p < 0.05 compared to the NC group; p < 0.01, p < 0.05 compared to the model group. PAE: P. americana extract; 5-ASA: Mesalazine, 200 mg/kg; PAE-L: 80 mg/kg, PAE-M: 160mg/kg, PAE-H: 320 mg/kg.

Article Snippet: Primary rabbit monoclonal antibodies against Nrf2 and b-tubulin were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Western Blot

Journal: Pharmaceutical biology

Article Title: Antioxidative effect of Periplaneta americana extract on dextran sulfate sodium-induced ulcerative colitis through activation of the Nrf2 signal.

doi: 10.1080/13880209.2023.2220351

Figure Lengend Snippet: Figure 9. PAE activated the Nrf2 signaling pathway in TNF-a-induced NCM460 cells. (A) Representative expression of GCLM, GPX2, HO-1, and NQO1. Relative expres- sion of GCLM (B), GPX2 (C), HO-1 (D), and NQO1 (E). Error bars represent the mean ± SEMs (n ¼ 3/group). ##p < 0.01, #p < 0.05 relative to NC group; p < 0.01, p < 0.05 relative to model group. PAE: P. americana extract, PAE-L: 400 lg/mL, PAE-M: 800 lg/mL, PAE-H: 1600 lg/mL; AA-L (3 lg/mL proline and 4 lg/mL glycine), AA-M (6 lg/mL proline and 8 lg/mL glycine), AA-H (12lg/mL proline and 16lg/mL glycine).

Article Snippet: Primary rabbit monoclonal antibodies against Nrf2 and b-tubulin were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing

Journal: Cells

Article Title: The Anti-Inflammatory Effect of the β1-Adrenergic Receptor Antagonist Metoprolol on High Glucose Treated Human Microvascular Retinal Endothelial Cells

doi: 10.3390/cells11010051

Figure Lengend Snippet: Anti ROS activity of metoprolol and Keap1/Nrf2/HO-1 axis activation in HREC that was treated with HG. The cells were treated in the presence of normal glucose levels (NG, 5 mM), or high glucose levels (HG, 25 mM) alone or supplemented with metoprolol (Met, 10 µM) and epinephrine (Epi, 1 µM) for 48 h. ( A ) ROS levels as evaluated by H2DCFDA assays. ( B ) Immunoblot analysis was performed using specific antibodies against Keap1, Nrf2, HO-1, and TNF proteins. The blots were probed with anti-GAPDH (reference) antibody to verify the equal loading of 30 µg protein per lane. Image J software was used to carry out densitometric analysis of the immunoblots, indicating protein quantification of each band (in arbitrary densitometry units, a.d.u.). The graphs show the target/GAPDH band intensity ratio that was normalized with respect to the corresponding reference for Keap1 ( C ), Nrf2 ( D ), HO-1 ( E ), and TNF-α ( F ). The data are representative of three independent experiments with four parallel samples for group in each experiment and are expressed as mean ± SD. * p < 0.05 vs. NG; # p < 0.05 vs. HG. The non-parametric Mann-Whitney test was used for pairwise comparisons.

Article Snippet: Rabbit polyclonal antibody against phospho-p44/42 MAPK (p-ERK1/2, catalog n. 9101), p44/42 MAPK (ERK1/2, catalog n. 9102), and GAPDH (catalog n. 2118) were purchased from Cell Signaling Technology (Danvers, MA, USA); mouse monoclonal antibody against Nrf2 (catalog n. ab137550) and mouse monoclonal antibody against TNF-α (catalog n. ab1793) were purchased from Abcam (Cambridge, UK).

Techniques: Activity Assay, Activation Assay, Western Blot, Software, MANN-WHITNEY

Journal: Cells

Article Title: The Anti-Inflammatory Effect of the β1-Adrenergic Receptor Antagonist Metoprolol on High Glucose Treated Human Microvascular Retinal Endothelial Cells

doi: 10.3390/cells11010051

Figure Lengend Snippet: The effects of metoprolol on Nrf2 nuclear translocation in HREC that was stimulated with high glucose levels. ( A ) The representative of immunocytochemically stained images for Nrf2 in HREC that was cultured in normal glucose levels (NG, 5 mM) or in HG, (25 mM) alone or supplemented with metoprolol (Met, 10 µM) for 48 h (in red: a′–c′ respectively). Moreover, the relative Hoechst-stained nuclei (a–c, respectively) and merged channels (a″–c″, respectively) are shown. All images were acquired with the Operetta High-Content Imaging System using a 20× magnification; scale bar = 100 µm. ( B ) High-content screening analysis was used to quantify the green fluorescence (Cy3) relative to the Nrf2 staining in the cytoplasm and nuclei. ( C ) Quantification of Nrf2 positive nuclei. The data are expressed as the mean ± SD from at least six fields/well randomly selected and each reporting more than 15 cells/field. All of the data represent the mean ± SD that were obtained from at least three independent experiments with four parallel samples for group in each experiment. * p < 0.05 vs. NG; # p < 0.05 vs. HG. The non-parametric Mann-Whitney test was used for pairwise comparisons.

Article Snippet: Rabbit polyclonal antibody against phospho-p44/42 MAPK (p-ERK1/2, catalog n. 9101), p44/42 MAPK (ERK1/2, catalog n. 9102), and GAPDH (catalog n. 2118) were purchased from Cell Signaling Technology (Danvers, MA, USA); mouse monoclonal antibody against Nrf2 (catalog n. ab137550) and mouse monoclonal antibody against TNF-α (catalog n. ab1793) were purchased from Abcam (Cambridge, UK).

Techniques: Translocation Assay, Staining, Cell Culture, Imaging, High Content Screening, Fluorescence, MANN-WHITNEY

Journal: Journal of Inflammation Research

Article Title: Sodium Propionate Enhances Nrf2-Mediated Protective Defense Against Oxidative Stress and Inflammation in Lipopolysaccharide-Induced Neonatal Mice

doi: 10.2147/JIR.S303105

Figure Lengend Snippet: Primers for Real-Time Quantitative PCR Analysis in Humans

Article Snippet: For Western blotting analysis, rabbit monoclonal antibody against Nrf2 was purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Real-time Polymerase Chain Reaction, Sequencing

Journal: Journal of Inflammation Research

Article Title: Sodium Propionate Enhances Nrf2-Mediated Protective Defense Against Oxidative Stress and Inflammation in Lipopolysaccharide-Induced Neonatal Mice

doi: 10.2147/JIR.S303105

Figure Lengend Snippet: SP treatment dampens inflammation and oxidative stress, amplifies Nrf2 activation and vascularization in LPS-treated neonatal mice lung. When the newborn mice were at day of life 6, they received intraperitoneal injection of 1 mg/kg LPS, whereas the control mice received an equal volume injection of sterile saline solution. The LPS group were then randomly divided into two groups that received vehicle or sodium propionate (1.2 mg/g) for 7 d. ( A ) Effect of SP on lung and heart organ index. ( B ) Effect of SP on the mRNA expressions of inflammatory cytokines. ( C and D ) Effect of SP on serum and lung SOD activity, respectively. ( E ) Effect of SP on the mRNA expressions of SOD1, SOD2, Gclm and Txn . ( F ) Representative images of Western blots. ( G ) The protein levels of Keap-1, Nrf2 and vWF. ( H ) Immunohistochemistry for vWF in the lung section. * P <0.05 vs saline; ** P <0.01 vs saline; *** P <0.001 vs saline; # P <0.05 vs LPS; ## P <0.01 vs LPS; ### P <0.001 vs LPS. Values are mean±SE, n=6 per group.

Article Snippet: For Western blotting analysis, rabbit monoclonal antibody against Nrf2 was purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Activation Assay, Injection, Control, Sterility, Saline, Activity Assay, Western Blot, Immunohistochemistry

Journal: Journal of Inflammation Research

Article Title: Sodium Propionate Enhances Nrf2-Mediated Protective Defense Against Oxidative Stress and Inflammation in Lipopolysaccharide-Induced Neonatal Mice

doi: 10.2147/JIR.S303105

Figure Lengend Snippet: SP prevents LPS-evoked pulmonary alveolar remodeling depending on Nrf2. When the newborn mice (WT and Nrf2 -/- ) were at day of life 6, they received intraperitoneal injection of 1 mg/kg LPS, whereas the control mice received an equal volume injection of sterile saline solution. The LPS group were then randomly divided into two groups that received vehicle or sodium propionate (1.2 mg/g) for 7 d. ( A ) The protein expression of Nrf2. ( B ) Representative H&E stained lung sections (magnification, ×200). ( C ) Quantification of MLI. ( D ) Lung and heart organ index. ( E ) Serum SOD activity. ( F ) The mRNA levels of inflammatory cytokines and Nrf2 target genes. * P <0.05 vs saline; ** P <0.01 vs saline; # P <0.05 vs LPS; ## P <0.01 vs LPS; n.s. vs no significance. Values are mean±SE, n=6 per group.

Article Snippet: For Western blotting analysis, rabbit monoclonal antibody against Nrf2 was purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Injection, Control, Sterility, Saline, Expressing, Staining, Activity Assay

Journal: Journal of Inflammation Research

Article Title: Sodium Propionate Enhances Nrf2-Mediated Protective Defense Against Oxidative Stress and Inflammation in Lipopolysaccharide-Induced Neonatal Mice

doi: 10.2147/JIR.S303105

Figure Lengend Snippet: SP blocks LPS-induced oxidative stress in HPMECs. HPMECs were stimulated with LPS (1 μg/mL) for 2 h, followed by treated with SP (0.6 mM) for 24 h. ( A ) Intracellular ROS was determined using DHE staining. ( B ) Relative fluorescent intensity of DHE staining. ( C ) The mRNA levels of antioxidant genes. ( D ) Representative Western Blots and quantification of Keap-1 and Nrf2. ( E ) Representative images of immunofluorescence staining of Nrf2 (green). Nuclei were stained with DAPI (blue). * P <0.05 vs PBS; ** P <0.01 vs PBS, *** P <0.001 vs PBS; # P <0.05 vs LPS; ## P <0.01 vs LPS. Values are mean±SE, n=3 per group.

Article Snippet: For Western blotting analysis, rabbit monoclonal antibody against Nrf2 was purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Staining, Western Blot, Immunofluorescence

Journal: Journal of Inflammation Research

Article Title: Sodium Propionate Enhances Nrf2-Mediated Protective Defense Against Oxidative Stress and Inflammation in Lipopolysaccharide-Induced Neonatal Mice

doi: 10.2147/JIR.S303105

Figure Lengend Snippet: SP blocks LPS-triggered HPMECs injury depending on Nrf2 activation. HPMECs were stimulated with ML358 (5 μM) for 1 h, followed by stimulated with LPS (1 μg/mL) for 2 h and treated with SP (0.6 mM) for additional 24 h. ( A ) mRNA levels of inflammatory cytokines. ( B ) mRNA levels of antioxidant genes. ( C ) The mRNA levels of angiogenesis-related genes. ( D ) Relative fluorescent intensity of DHE staining. ( E ) Representative DHE staining images. # P <0.05 vs LPS; ## P <0.01 vs LPS; ### P <0.001 vs LPS; † P <0.05 vs LPS + SP; †† P <0.01 vs LPS + SP; n.s. vs no significance. Values are mean ± SE, n=3 per group.

Article Snippet: For Western blotting analysis, rabbit monoclonal antibody against Nrf2 was purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Activation Assay, Staining