rabbit polyclonal antibody against protein nrf2  (Proteintech)

Journal: Drug Design, Development and Therapy

Article Title: Systematic Pharmacology and Experimental Validation to Reveal the Alleviation of Astragalus membranaceus Regulating Ferroptosis in Osteoarthritis

doi: 10.2147/DDDT.S441350

Figure Lengend Snippet: Molecular interaction of Astragalus membranaceus with target proteins. ( A ) Heatmap of CB-DOCK scores for the main active components of Astragalus membranaceus and screened targets. ( B ) Histogram of the number of targets of the main active components of Astragalus membranaceus. ( C ) The 3D model of the interaction between Quercetin and the target protein with 6 highest docking energy. ( D ) Domain structure of Nrf2.

Article Snippet: The cells were then incubated with a rabbit polyclonal antibody against protein NRF2 (1:500 dilution, Proteintech, Wuhan, China) overnight at 4°C.

Techniques:

Journal: Drug Design, Development and Therapy

Article Title: Systematic Pharmacology and Experimental Validation to Reveal the Alleviation of Astragalus membranaceus Regulating Ferroptosis in Osteoarthritis

doi: 10.2147/DDDT.S441350

Figure Lengend Snippet: Quercetin alleviated FAC-induced ferroptosis in chondrocytes via NRF2/KEAP1 pathway. ( A ) Protein analysis of NRF2, KEAP1, p38, p-p38, HO-1 and GAPDH, and quantification of NRF2, KEAP1, p38, p-p38 and HO-1. ( B ) Immunofluorescence of NRF2 showing nuclear translocation of NRF2 under different conditions. Blue: DAPI; Green: NRF2. ( C ) Mean Manders’ Colocalization Coefficients of NRF2 (n = 5). ( D ) Mean fluorescence intensity of NRF2 (n = 5). ( E ) Viability curves of chondrocytes under different conditions. ( F ) Immunoprecipitation results. Immunoprecipitation was performed using the Anti-KEAP1 antibody, followed by Western blot analysis of KEAP1 and NRF2. Data represent the mean ± SD.

Article Snippet: The cells were then incubated with a rabbit polyclonal antibody against protein NRF2 (1:500 dilution, Proteintech, Wuhan, China) overnight at 4°C.

Techniques: Immunofluorescence, Translocation Assay, Fluorescence, Immunoprecipitation, Western Blot

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Activation of Nrf2 inhibits ferroptosis and protects against oxaliplatin-induced ototoxicity.

doi: 10.1016/j.biopha.2023.115248

Figure Lengend Snippet: Fig. 1. Oxaliplatin inhibited OC1 cells viability and increased the level of lipid peroxidation. (A) OC1 cells treated with varying concentrations of oxaliplatin for 24 h and cell viability were analyzed by CCK-8. (B) The relative levels of MDA in OC1 cells treated with oxaliplatin at different concentrations for 24 h. (C) The relative level of SOD in OC1 cells treated with oxaliplatin at different concentrations for 24 h. (D) Intracellular ROS levels detected by DCFH, and 4HNE, GSH immunostaining was used to detect lipid peroxidation level. (E) The quantification of 4HNE expression level in OC-1 cells treated with oxaliplatin at different concentrations. (F) The quantification of GSH expression level. (G) Representative western blots of GPX4, Nrf2, and HO-1 expression in HEI-OC1 cells of each group. (H) The quantification of GPX4 expression level. (I) The quantification of Nrf2 expression level. (G) The quantification of HO-1 expression level. * P < 0.05, ** P < 0.01, and *** P < 0.001. Scale in panels D represents for 50 µm.

Article Snippet: The membranes were blocked in Tris-buffered saline (TBST with 0.1 % Tween-20) containing 5 % milk for 1.5 h. The expression of Nrf2, HO-1, GPX4 and IRP-2 were detected using rabbit polyclonal antibodies against Nrf2 (20733S, Cell Signaling Technology, Danvers, MA, USA), rabbit HO-1 polyclonal K. Xu et al. Biomedicine & Pharmacotherapy 165 (2023) 115248 antibody (10701-1-AP, Proteintech, Rosemont, IL, USA), mouse GPX4 monoclonal antibody (67763-1-Ig, Proteintech), mouse monoclonal anti-IRP-2 antibody (sc-33680; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit β-actin recombinant antibody (81115-1-RR, Proteintech).

Techniques: CCK-8 Assay, Immunostaining, Expressing, Western Blot

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Activation of Nrf2 inhibits ferroptosis and protects against oxaliplatin-induced ototoxicity.

doi: 10.1016/j.biopha.2023.115248

Figure Lengend Snippet: Fig. 3. Inhibition of ferroptosis with ferrostatin-1 ameliorated oxaliplatin induced cytotoxicity in OC1 cells. ( A) The cell viability of OC1 cells in different treatment groups. (B) Flow cytometry analysis apoptosis of OC1 cells in different treatment groups. (C) Quantitative apoptosis of OC-1 cells in each group. (D) DCFH staining and flow cytometry analysis intracellular ROS levels of OC1 cells in different treatment groups. (E) Western blot analysis of GPX4, Nrf2, and HO-1 levels in OC1 cells in different treatment groups. (F–H) The quantification of GPX4, Nrf2, and HO-1expression level in different treatment groups. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: The membranes were blocked in Tris-buffered saline (TBST with 0.1 % Tween-20) containing 5 % milk for 1.5 h. The expression of Nrf2, HO-1, GPX4 and IRP-2 were detected using rabbit polyclonal antibodies against Nrf2 (20733S, Cell Signaling Technology, Danvers, MA, USA), rabbit HO-1 polyclonal K. Xu et al. Biomedicine & Pharmacotherapy 165 (2023) 115248 antibody (10701-1-AP, Proteintech, Rosemont, IL, USA), mouse GPX4 monoclonal antibody (67763-1-Ig, Proteintech), mouse monoclonal anti-IRP-2 antibody (sc-33680; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit β-actin recombinant antibody (81115-1-RR, Proteintech).

Techniques: Inhibition, Flow Cytometry, Staining, Western Blot

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Activation of Nrf2 inhibits ferroptosis and protects against oxaliplatin-induced ototoxicity.

doi: 10.1016/j.biopha.2023.115248

Figure Lengend Snippet: Fig. 4. Oxaliplatin caused degeneration of hair cell in murine cochlear explants. (A) Experimental workflow of cochlear explants culture. (B) Representative images of cochlear hair cells (green) of different turns following treated with different concentrations of oxaliplatin. (C) Quantified the number of HCs at specific cochlear locations from different group. (D) Total amount of cochlear GPX4 and IRP-2 protein levels following treated with oxaliplatin were assessed by western blotting. (E, F) The representative quantification of GPX4 and IRP-2 protein expression from western blotting. (G) Western blot analysis of Nrf2 and HO-1 levels in each group. (H, I) The representative quantification of Nrf2 and HO-1 protein expression from western blotting. * P < 0.05, ** P < 0.01, and *** P < 0.001. Scale in panels B represents for 40 µm.

Article Snippet: The membranes were blocked in Tris-buffered saline (TBST with 0.1 % Tween-20) containing 5 % milk for 1.5 h. The expression of Nrf2, HO-1, GPX4 and IRP-2 were detected using rabbit polyclonal antibodies against Nrf2 (20733S, Cell Signaling Technology, Danvers, MA, USA), rabbit HO-1 polyclonal K. Xu et al. Biomedicine & Pharmacotherapy 165 (2023) 115248 antibody (10701-1-AP, Proteintech, Rosemont, IL, USA), mouse GPX4 monoclonal antibody (67763-1-Ig, Proteintech), mouse monoclonal anti-IRP-2 antibody (sc-33680; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit β-actin recombinant antibody (81115-1-RR, Proteintech).

Techniques: Western Blot, Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Activation of Nrf2 inhibits ferroptosis and protects against oxaliplatin-induced ototoxicity.

doi: 10.1016/j.biopha.2023.115248

Figure Lengend Snippet: Fig. 5. Effects of ferroptosis and RSL3 on HCs damaged induced by oxaliplatin. (A) Representative images of cochlear hair cells (green) of different turns from different treatment group. (B) Quantified the number of HCs at specific cochlear locations from different group. (C) The expression levels of GPX4, IRP-2, Nrf2 and HO-1 in cochlea from different group were assessed by western blotting. (D–G) The representative quantification of GPX4, IRP-2, Nrf2 and HO-1 protein expression in cochlea from western blotting. * P < 0.05, ** P < 0.01, and *** P < 0.001. Scale in panels A represents for 40 µm.

Article Snippet: The membranes were blocked in Tris-buffered saline (TBST with 0.1 % Tween-20) containing 5 % milk for 1.5 h. The expression of Nrf2, HO-1, GPX4 and IRP-2 were detected using rabbit polyclonal antibodies against Nrf2 (20733S, Cell Signaling Technology, Danvers, MA, USA), rabbit HO-1 polyclonal K. Xu et al. Biomedicine & Pharmacotherapy 165 (2023) 115248 antibody (10701-1-AP, Proteintech, Rosemont, IL, USA), mouse GPX4 monoclonal antibody (67763-1-Ig, Proteintech), mouse monoclonal anti-IRP-2 antibody (sc-33680; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit β-actin recombinant antibody (81115-1-RR, Proteintech).

Techniques: Expressing, Western Blot

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Activation of Nrf2 inhibits ferroptosis and protects against oxaliplatin-induced ototoxicity.

doi: 10.1016/j.biopha.2023.115248

Figure Lengend Snippet: Fig. 6. Effects of resveratrol on HCs damaged induced by oxaliplatin. (A) Total amount of cochlear Nrf2 expression levels following treated with resveratrol were assessed by western blotting. (B) Quantification of Nrf2 expression in cochlea. (C, D) The expression of Nrf2 in nuclear were analyzed by western blot, and quantification of the Nrf2 protein levels by normalized to Histone-H3. (E) Total amount of cochlear GPX4 and IRP-2 expression levels from different group were assessed by western blotting. (F, G) Quantification of GPX4 and IRP-2 protein expression in cochlea from western blotting. (H) Representative images of cochlear hair cells (green) of different turns from different treatment group. (I) Quantified the number of HCs at specific cochlear locations from different group. * P < 0.05, ** P < 0.01, and *** P < 0.001. Scale in panels H represents for 40 µm.

Article Snippet: The membranes were blocked in Tris-buffered saline (TBST with 0.1 % Tween-20) containing 5 % milk for 1.5 h. The expression of Nrf2, HO-1, GPX4 and IRP-2 were detected using rabbit polyclonal antibodies against Nrf2 (20733S, Cell Signaling Technology, Danvers, MA, USA), rabbit HO-1 polyclonal K. Xu et al. Biomedicine & Pharmacotherapy 165 (2023) 115248 antibody (10701-1-AP, Proteintech, Rosemont, IL, USA), mouse GPX4 monoclonal antibody (67763-1-Ig, Proteintech), mouse monoclonal anti-IRP-2 antibody (sc-33680; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit β-actin recombinant antibody (81115-1-RR, Proteintech).

Techniques: Expressing, Western Blot

Journal: Biology

Article Title: Cepabiflas B and C as Novel Anti-Inflammatory and Anti-Apoptotic Agents against Endotoxin-Induced Acute Kidney and Hepatic Injury in Mice: Impact on Bax/Bcl2 and Nrf2/NF-κB Signalling Pathways

doi: 10.3390/biology12070938

Figure Lengend Snippet: The sequence of the primers employed in RT-PCR.

Article Snippet: Sections were IHC stained using the primary antibodies: rabbit-polyclonal-antibody against NF-kB p65 (1:200), Nrf2 (1:200) (Fisher-Scientific Inc., Waltham, MA, USA), Bcl2 (1:200), and caspase-3 (1:200) (Elabscience Biotechnology Inc., Houston, TX, USA).

Techniques: Sequencing

Journal: Biology

Article Title: Cepabiflas B and C as Novel Anti-Inflammatory and Anti-Apoptotic Agents against Endotoxin-Induced Acute Kidney and Hepatic Injury in Mice: Impact on Bax/Bcl2 and Nrf2/NF-κB Signalling Pathways

doi: 10.3390/biology12070938

Figure Lengend Snippet: CBs enhanced Nrf2 signalling in the liver and the kidney of LPS-intoxicated mice. ( A , B ) mRNA expression of Nrf2 and its binding activity. ( C , D ) mRNA expression and level of HO-1 in hepatic and kidney tissues. ( E ) Immuno-expression of Nrf2 in the liver and kidney tissue where specimen of LPS group showed decreased immuno-stain compared to the control group while specimen of CBs pre-treated groups exhibited enhanced Nrf2 immuno-stain. ( F ) % of immuno-positive cells of Nrf2. Data are the mean ± SE (n = 6). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group; p < 0.05, ## p < 0.01, ### p < 0.001 vs. LPS group (one-way ANOVA).

Article Snippet: Sections were IHC stained using the primary antibodies: rabbit-polyclonal-antibody against NF-kB p65 (1:200), Nrf2 (1:200) (Fisher-Scientific Inc., Waltham, MA, USA), Bcl2 (1:200), and caspase-3 (1:200) (Elabscience Biotechnology Inc., Houston, TX, USA).

Techniques: Expressing, Binding Assay, Activity Assay, Immunostaining

Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Hepatoprotective and MicroRNAs Downregulatory Effects of Crocin Following Hepatic Ischemia-Reperfusion Injury in Rats

doi: 10.1155/2017/1702967

Figure Lengend Snippet: Immunohistochemical staining of liver Nrf2 expression (immunohistochemistry, ×300). Sham group: Nrf2 localized predominantly in cytoplasm of hepatocytes. IR group: Nrf2 expression decreased in cytoplasm and moderately increased in nucleus. Cr group: The immune reaction was moderate in cytoplasm. Cr + IR group: Nrf2 expression increased both in nucleus and in cytoplasm, but its expression was in nucleus higher than in cytoplasm.

Article Snippet: Thereafter, sections were incubated with rabbit polyclonal antibody against Nrf2 (dilution 1 : 100; Abcam [ab31163], USA) and then incubated with a secondary antibody (dilution 1 : 1000; Abcam [ab6721], USA), in a moist chamber.

Techniques: Immunohistochemical staining, Staining, Expressing, Immunohistochemistry