Structured Review

Abcam anti nrf2
Summary of the hypothesized interaction within the <t>estradiol–ERα–NRF2</t> pathway: High expression of ERα leads to an induction of the estradiol–ERα-dependent pathway, resulting in transcriptionally inactive NRF2 (low nuclear, high cytoplasmic expression) and consecutively less impact on tumor growth. In contrast, low ERα expression favors the estradiol–ERα-independent pathway, with activation of NRF2 (high nuclear, low cytoplasmic expression) causing tumor progression.
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Images

1) Product Images from "Interaction of ERα and NRF2 Impacts Survival in Ovarian Cancer Patients"

Article Title: Interaction of ERα and NRF2 Impacts Survival in Ovarian Cancer Patients

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms20010112

Summary of the hypothesized interaction within the estradiol–ERα–NRF2 pathway: High expression of ERα leads to an induction of the estradiol–ERα-dependent pathway, resulting in transcriptionally inactive NRF2 (low nuclear, high cytoplasmic expression) and consecutively less impact on tumor growth. In contrast, low ERα expression favors the estradiol–ERα-independent pathway, with activation of NRF2 (high nuclear, low cytoplasmic expression) causing tumor progression.
Figure Legend Snippet: Summary of the hypothesized interaction within the estradiol–ERα–NRF2 pathway: High expression of ERα leads to an induction of the estradiol–ERα-dependent pathway, resulting in transcriptionally inactive NRF2 (low nuclear, high cytoplasmic expression) and consecutively less impact on tumor growth. In contrast, low ERα expression favors the estradiol–ERα-independent pathway, with activation of NRF2 (high nuclear, low cytoplasmic expression) causing tumor progression.

Techniques Used: Expressing, Activation Assay

2) Product Images from "Interaction of ERα and NRF2 Impacts Survival in Ovarian Cancer Patients"

Article Title: Interaction of ERα and NRF2 Impacts Survival in Ovarian Cancer Patients

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms20010112

Summary of the hypothesized interaction within the estradiol–ERα–NRF2 pathway: High expression of ERα leads to an induction of the estradiol–ERα-dependent pathway, resulting in transcriptionally inactive NRF2 (low nuclear, high cytoplasmic expression) and consecutively less impact on tumor growth. In contrast, low ERα expression favors the estradiol–ERα-independent pathway, with activation of NRF2 (high nuclear, low cytoplasmic expression) causing tumor progression.
Figure Legend Snippet: Summary of the hypothesized interaction within the estradiol–ERα–NRF2 pathway: High expression of ERα leads to an induction of the estradiol–ERα-dependent pathway, resulting in transcriptionally inactive NRF2 (low nuclear, high cytoplasmic expression) and consecutively less impact on tumor growth. In contrast, low ERα expression favors the estradiol–ERα-independent pathway, with activation of NRF2 (high nuclear, low cytoplasmic expression) causing tumor progression.

Techniques Used: Expressing, Activation Assay

Correlation analysis of NRF2 and ERα in ovarian cancer tissue ( n = 139). A significant correlation of cytoplasmic NRF2 expression with ERα expression was noted. For better visualization, dots have been jittered.
Figure Legend Snippet: Correlation analysis of NRF2 and ERα in ovarian cancer tissue ( n = 139). A significant correlation of cytoplasmic NRF2 expression with ERα expression was noted. For better visualization, dots have been jittered.

Techniques Used: Expressing

Kaplan–Meier estimates of NRF2 expression, ERα expression, and combined NRF2 and ERα expression were analyzed. In the serous subtype, patients with a high cytoplasmic expression of NRF2 showed a significantly increased overall survival compared with patients with a low cytoplasmic expression ( A ). In addition, high ERα expression was associated with significantly better overall survival in serous ovarian cancer compared with patients with a low ERα expression ( B ). Patients with combined high NRF2 expression in the cytoplasm and ERα expression in epithelial ovarian carcinoma (EOC) had significantly increased overall survival compared with those with low cytoplasmic expression and ERα expression ( C ).
Figure Legend Snippet: Kaplan–Meier estimates of NRF2 expression, ERα expression, and combined NRF2 and ERα expression were analyzed. In the serous subtype, patients with a high cytoplasmic expression of NRF2 showed a significantly increased overall survival compared with patients with a low cytoplasmic expression ( A ). In addition, high ERα expression was associated with significantly better overall survival in serous ovarian cancer compared with patients with a low ERα expression ( B ). Patients with combined high NRF2 expression in the cytoplasm and ERα expression in epithelial ovarian carcinoma (EOC) had significantly increased overall survival compared with those with low cytoplasmic expression and ERα expression ( C ).

Techniques Used: Expressing

Detection of nuclear factor erythroid-2-related factor 2 (NRF2) ( A1, B1 ) and estrogen receptor (ER)α ( A2, B2 ) with immunohistochemistry. High ( A1 ) and low ( B1 ) cytoplasmic NRF2 stains in serous subtype correspond with high ( A2 ) and low ( B2 ) ERα stains, respectively. NRF2 shows faint staining in the nucleus in both cases ( A1, B1 ).
Figure Legend Snippet: Detection of nuclear factor erythroid-2-related factor 2 (NRF2) ( A1, B1 ) and estrogen receptor (ER)α ( A2, B2 ) with immunohistochemistry. High ( A1 ) and low ( B1 ) cytoplasmic NRF2 stains in serous subtype correspond with high ( A2 ) and low ( B2 ) ERα stains, respectively. NRF2 shows faint staining in the nucleus in both cases ( A1, B1 ).

Techniques Used: Immunohistochemistry, Staining

3) Product Images from "A phenolic amide (LyA) isolated from the fruits of Lycium barbarum protects against cerebral ischemia–reperfusion injury via PKCε/Nrf2/HO-1 pathway"

Article Title: A phenolic amide (LyA) isolated from the fruits of Lycium barbarum protects against cerebral ischemia–reperfusion injury via PKCε/Nrf2/HO-1 pathway

Journal: Aging (Albany NY)

doi: 10.18632/aging.102578

LyA Promoted the Expression of Nrf2 and HO-1. ( A ) Protein expressions of Nuclear Nrf2, Cytoplasmic Nrf2 and HO-1 were evaluated by Western blot analysis. ( B ) The immunofluorescence staining of Nrf2 with DAPI (400 x). Data were presented as mean ± SD (n = 6). * p
Figure Legend Snippet: LyA Promoted the Expression of Nrf2 and HO-1. ( A ) Protein expressions of Nuclear Nrf2, Cytoplasmic Nrf2 and HO-1 were evaluated by Western blot analysis. ( B ) The immunofluorescence staining of Nrf2 with DAPI (400 x). Data were presented as mean ± SD (n = 6). * p

Techniques Used: Expressing, Western Blot, Immunofluorescence, Staining

PKCε mediated LyA-induced neuroprotection, Nrf2 nuclear translocation, and HO-1 upregulation. ( A – C ) Cells were transfected with control or PKCε siRNA for 48 h, followed by treatment with 40 μM LyA for 8 h. p-PKCε, PKCε, HO-1, Nrf2, bax, bcl-2 and cleaved caspase-3 expression levels were analyzed by western blotting. Data were presented as mean ± SD (n =6). * p
Figure Legend Snippet: PKCε mediated LyA-induced neuroprotection, Nrf2 nuclear translocation, and HO-1 upregulation. ( A – C ) Cells were transfected with control or PKCε siRNA for 48 h, followed by treatment with 40 μM LyA for 8 h. p-PKCε, PKCε, HO-1, Nrf2, bax, bcl-2 and cleaved caspase-3 expression levels were analyzed by western blotting. Data were presented as mean ± SD (n =6). * p

Techniques Used: Translocation Assay, Transfection, Expressing, Western Blot

Effect of LyA treatment on PKCε phosphorylation, activation of Nrf2/HO-1 pathway in SH-SY5Y cells. ( A ) Representative bands of each protein are presented. ( B ) Relative expressions are calculated and are shown here. Data were represented as means ± SD (n=6). * p
Figure Legend Snippet: Effect of LyA treatment on PKCε phosphorylation, activation of Nrf2/HO-1 pathway in SH-SY5Y cells. ( A ) Representative bands of each protein are presented. ( B ) Relative expressions are calculated and are shown here. Data were represented as means ± SD (n=6). * p

Techniques Used: Activation Assay

The neuroprotection of LyA involved the Nrf2/HO-1 pathway. ( A – C ) Cells were transfected with control or HO-1 and Nrf2 siRNA for 48 h, followed by treatment with 40 μM LyA for 8 h. HO-1 and Nrf2 expression levels were analyzed by western blotting. Data were presented as mean ± SD (n =6). * p
Figure Legend Snippet: The neuroprotection of LyA involved the Nrf2/HO-1 pathway. ( A – C ) Cells were transfected with control or HO-1 and Nrf2 siRNA for 48 h, followed by treatment with 40 μM LyA for 8 h. HO-1 and Nrf2 expression levels were analyzed by western blotting. Data were presented as mean ± SD (n =6). * p

Techniques Used: Transfection, Expressing, Western Blot

Related Articles

Blocking Assay:

Article Title: Tert-butylhydroquinone attenuates the ethanol-induced apoptosis of and activates the Nrf2 antioxidant defense pathway in H9c2 cardiomyocytes
Article Snippet: The cells were then permeabilized with 0.1% Triton X-100, washed and incubated with blocking buffer (10% NGS) for 1 h at room temperature. .. The cells were then incubated overnight with primary antibodies against AC-tubulin (6-11B-1; ab24610) at 1:200 dilution and Nrf2 (both from Abcam) at a 1:200 dilution in a humidified chamber at 4°C.

Article Title: Hormetic potential of methylglyoxal, a side-product of glycolysis, in switching tumours from growth to death
Article Snippet: .. Blocking was performed in 5% non-fat dried milk (Biorad, Hercules, CA) in TBS-Tween 0.1% for 1 h. Membranes were then incubated overnight at 4 °C with primary antibodies: Argpyrimidine (1/10,000, mAb6B), MG-H1 (1/1000, STA-011, Cell Biolabs), Nrf2 (1/1000, Ab62353, Abcam), GLO1 (1/1000, #02-14, BioMac, Leipzig, Germany), β-actin (1/5000, A5441, Sigma). .. Membranes were then incubated for 1 h in the presence of the appropriate secondary antibody coupled to horseradish peroxidase.

Electrophoresis:

Article Title: Brain ischemic preconditioning protects against ischemic injury and preserves the blood-brain barrier via oxidative signaling and Nrf2 activation
Article Snippet: Equal amounts of protein samples were loaded onto acrylamide gels and subjected to electrophoresis, followed by transfer to polyvinylidene difluoride (PVDF) membranes (Bio-Rad). .. PVDF membranes were then blocked with 5% non-fat milk, and probed with antibodies recognizing hemo oxygenase-1 (HO-1, 1:1000, Enzo Life Science, Farmingdale, NY), Nrf2 (1:1000, Abcam, Cambridge, MA), glutamate-cysteine ligase modifier subunit (GCLM, 1:1000, Abcam), β-actin (1:3000, Sigma-Aldrich, St. Louis, MO), FLAG (1:1000, Cell Signaling Technology, Danvers, MA), cadherin 5 (CDH5, 1:1000, Abcam), claudin 5 (1:1000, Abcam), Akt (1:1000, Cell Signaling Technology), phosphorylated Akt (p-Akt, S473, 1:1000, Cell Signaling Technology), GSK3β (1:1000, Cell Signaling Technology), phosphorylated GSK3β (p-GSK3β, S9, 1:1000, Cell Signaling Technology), human influenza hemagglutinin (HA, 1:1000, Cell Signaling Technology) or lamin B1 (1:1000, Cell Signaling Technology).

Incubation:

Article Title: Tert-butylhydroquinone attenuates the ethanol-induced apoptosis of and activates the Nrf2 antioxidant defense pathway in H9c2 cardiomyocytes
Article Snippet: .. The cells were then incubated overnight with primary antibodies against AC-tubulin (6-11B-1; ab24610) at 1:200 dilution and Nrf2 (both from Abcam) at a 1:200 dilution in a humidified chamber at 4°C. .. Subsequently, the cells were washed with PBS and incubated with secondary antibodies conjugated to either FITC or TRITC at a dilution of 1:500 (Jackson ImmunoResearch Laboratories, Inc., West Grove, PA, USA) for 1 h at room temperature in the dark, and were then counterstained with DAPI dye to show the nuclear morphology.

Article Title: Brain ischemic preconditioning protects against ischemic injury and preserves the blood-brain barrier via oxidative signaling and Nrf2 activation
Article Snippet: PVDF membranes were then blocked with 5% non-fat milk, and probed with antibodies recognizing hemo oxygenase-1 (HO-1, 1:1000, Enzo Life Science, Farmingdale, NY), Nrf2 (1:1000, Abcam, Cambridge, MA), glutamate-cysteine ligase modifier subunit (GCLM, 1:1000, Abcam), β-actin (1:3000, Sigma-Aldrich, St. Louis, MO), FLAG (1:1000, Cell Signaling Technology, Danvers, MA), cadherin 5 (CDH5, 1:1000, Abcam), claudin 5 (1:1000, Abcam), Akt (1:1000, Cell Signaling Technology), phosphorylated Akt (p-Akt, S473, 1:1000, Cell Signaling Technology), GSK3β (1:1000, Cell Signaling Technology), phosphorylated GSK3β (p-GSK3β, S9, 1:1000, Cell Signaling Technology), human influenza hemagglutinin (HA, 1:1000, Cell Signaling Technology) or lamin B1 (1:1000, Cell Signaling Technology). .. After incubation in secondary antibodies (Santa Cruz, Santa Cruz, CA) for 1 h, the membranes were incubated with ECL substrates (Pierce, ThermoFisher Scientific) and developed with X-ray film.

Article Title: KEAP1 Is a Redox Sensitive Target That Arbitrates the Opposing Radiosensitive Effects of Parthenolide in Normal and Cancer Cells
Article Snippet: .. Homogenized cells and tumor tissues were electrophoresed on an 8% (w/v) SDS-PAGE gel, transferred onto a nitrocellulose membrane, and subsequently incubated with primary antibodies against Keap1 (abcam), Nrf2 (abcam), MnSOD (Upstate Biotech), CuZnSOD (eBiosci), Gpx (abcam), catalase (Millipore), TrX (BD Sciences), PGAM5 (Santa Cruz Biotech.), Bcl-xL (Santa Cruz Biotech.), LC3B (cell signaling), β-actin (sigma), and pCNA (Santa Cruz Biotech.). ..

Article Title: Hormetic potential of methylglyoxal, a side-product of glycolysis, in switching tumours from growth to death
Article Snippet: .. Blocking was performed in 5% non-fat dried milk (Biorad, Hercules, CA) in TBS-Tween 0.1% for 1 h. Membranes were then incubated overnight at 4 °C with primary antibodies: Argpyrimidine (1/10,000, mAb6B), MG-H1 (1/1000, STA-011, Cell Biolabs), Nrf2 (1/1000, Ab62353, Abcam), GLO1 (1/1000, #02-14, BioMac, Leipzig, Germany), β-actin (1/5000, A5441, Sigma). .. Membranes were then incubated for 1 h in the presence of the appropriate secondary antibody coupled to horseradish peroxidase.

Article Title: Cerebroprotection by salvianolic acid B after experimental subarachnoid hemorrhage occurs via Nrf2- and SIRT1-dependent pathways
Article Snippet: .. Briefly, brain sections (6 μm) or cultured cells were incubated with primary antibodies against 8-hydroxyguanosine (8-OHdG, Abcam), Nrf2 (1:100, Abcam), SIRT1 (1:50, Santa Cruz), caspase-3 (1:100, Cell Signaling), and NeuN (1:200, cat# MAB377, EMD Millipore, USA) overnight at 4 °C. .. Brain sections were then incubated with corresponding secondary antibodies Alexa Fluor 488 and/or Alexa Fluor 594 (Jackson ImmunoResearch Incorporation, West Grove, PA, USA).

Article Title: T-type calcium channel blocker attenuates unilateral ureteral obstruction-induced renal interstitial fibrosis by activating the Nrf2 antioxidant pathway
Article Snippet: Primary antibodies against the following proteins were used: Nox1 and Nox4 (Santa Cruz Biotechnology), superoxide dismutase (SOD) 1 and 2 (Abcam), heme oxygenase (HO)-1 (BD Biosciences, San Jose, CA, USA), NAD(P)H: quinone oxidoreductase-1 (NQO-1; Santa Cruz Biotechnology), catalase (Abcam), B-cell lymphoma 2 (Bcl-2; Santa Cruz Biotechnology), Kelch-like ECH-associated protein 1 (Keap1; Abcam), Nrf2 (Abcam), and p300/CBP-associated factor (PCAF; Santa Cruz Biotechnology). .. The blots were washed and incubated with a secondary antibody conjugated with horseradish peroxidase (Jackson ImmunoResearch Laboratories, West Grove, PA, USA).

Article Title: TCF7L1 indicates prognosis and promotes proliferation through activation of Keap1/NRF2 in gastric cancer
Article Snippet: Subsequently, the membrane was incubated with indicated primary antibodies followed by washing with TBST and incubation with secondary antibody that conjugated with Horseradish Peroxidase (HRP). .. Antibodies against TCF7L1 and NRF2 were purchased from Abcam (Cambridge, UK).

Article Title: Oral Monomethyl Fumarate Therapy Ameliorates Retinopathy in a Humanized Mouse Model of Sickle Cell Disease
Article Snippet: .. Protein samples were subjected to SDS–PAGE, transferred to nitrocellulose membranes, and then incubated with primary antibodies: HbF (1:1000; Bethyl Laboratories, Inc.), Nrf2 (1:1000; AbCam), ZO-1 (1:100; Abcam), and VEGF (1:100; Abcam) overnight at 4°C. .. Detection of HbF, Nrf2, and ZO-1 was done using horseradish peroxidase-conjugated secondary antibodies.

Expressing:

Article Title: Carnosol as a Nrf2 Activator Improves Endothelial Barrier Function Through Antioxidative Mechanisms
Article Snippet: .. Assessment Expression of HO-1 and Nrf2 Proteins Antibodies for HO-1, β-actin were purchased from Proteintech (Rosemont, IL, USA), antibody for Nrf2 was purchased from Abcam (Cambridge, UK). ..

BIA-KA:

Article Title: TCF7L1 indicates prognosis and promotes proliferation through activation of Keap1/NRF2 in gastric cancer
Article Snippet: Protein concentrations of the whole cell lysate were measured by using BCA Protein Assay kit (Thermo Pierce, Rockford, USA). .. Antibodies against TCF7L1 and NRF2 were purchased from Abcam (Cambridge, UK).

Western Blot:

Article Title: Brain ischemic preconditioning protects against ischemic injury and preserves the blood-brain barrier via oxidative signaling and Nrf2 activation
Article Snippet: Paragraph title: Western blots and western dot blots ... PVDF membranes were then blocked with 5% non-fat milk, and probed with antibodies recognizing hemo oxygenase-1 (HO-1, 1:1000, Enzo Life Science, Farmingdale, NY), Nrf2 (1:1000, Abcam, Cambridge, MA), glutamate-cysteine ligase modifier subunit (GCLM, 1:1000, Abcam), β-actin (1:3000, Sigma-Aldrich, St. Louis, MO), FLAG (1:1000, Cell Signaling Technology, Danvers, MA), cadherin 5 (CDH5, 1:1000, Abcam), claudin 5 (1:1000, Abcam), Akt (1:1000, Cell Signaling Technology), phosphorylated Akt (p-Akt, S473, 1:1000, Cell Signaling Technology), GSK3β (1:1000, Cell Signaling Technology), phosphorylated GSK3β (p-GSK3β, S9, 1:1000, Cell Signaling Technology), human influenza hemagglutinin (HA, 1:1000, Cell Signaling Technology) or lamin B1 (1:1000, Cell Signaling Technology).

Article Title: Apurinic/Apyrimidinic Endonuclease/Redox Factor-1 (APE1/Ref-1) Redox Function Negatively Regulates NRF2 *
Article Snippet: Paragraph title: Western Blot Analysis ... Proteins were separated by SDS-PAGE, electroblotted onto nitrocellulose, and immunoblotting was performed using the following antibodies: Ref-1 and NRF2 (Abcam 62352), HMOX-1 (Abcam, Cambridge, MA), and Ku70, tubulin, or Actin (Sigma).

Article Title: KEAP1 Is a Redox Sensitive Target That Arbitrates the Opposing Radiosensitive Effects of Parthenolide in Normal and Cancer Cells
Article Snippet: Paragraph title: Immunoblots and immunoprecipitation ... Homogenized cells and tumor tissues were electrophoresed on an 8% (w/v) SDS-PAGE gel, transferred onto a nitrocellulose membrane, and subsequently incubated with primary antibodies against Keap1 (abcam), Nrf2 (abcam), MnSOD (Upstate Biotech), CuZnSOD (eBiosci), Gpx (abcam), catalase (Millipore), TrX (BD Sciences), PGAM5 (Santa Cruz Biotech.), Bcl-xL (Santa Cruz Biotech.), LC3B (cell signaling), β-actin (sigma), and pCNA (Santa Cruz Biotech.).

Article Title: Peroxiredoxin I is important for cancer-cell survival in Ras-induced hepatic tumorigenesis
Article Snippet: Paragraph title: Western blotting ... The membranes were primarily blotted with primary antibodies against Prx I, PCNA, Nrf2 (Abcam, Cambridge, UK); HA (Roche, Basel, Switzerland); GAPDH, FoxM1 (Abfrontier, Seoul, Korea); pERK, pP38, pJNK, cyclin D1 (Cell Signaling Technology, Danvers, MA, USA); and H-ras (Santa Cruz Biotechnology, Dallas, TX, USA).

Article Title: Hormetic potential of methylglyoxal, a side-product of glycolysis, in switching tumours from growth to death
Article Snippet: Paragraph title: Western Blot ... Blocking was performed in 5% non-fat dried milk (Biorad, Hercules, CA) in TBS-Tween 0.1% for 1 h. Membranes were then incubated overnight at 4 °C with primary antibodies: Argpyrimidine (1/10,000, mAb6B), MG-H1 (1/1000, STA-011, Cell Biolabs), Nrf2 (1/1000, Ab62353, Abcam), GLO1 (1/1000, #02-14, BioMac, Leipzig, Germany), β-actin (1/5000, A5441, Sigma).

Article Title: T-type calcium channel blocker attenuates unilateral ureteral obstruction-induced renal interstitial fibrosis by activating the Nrf2 antioxidant pathway
Article Snippet: Paragraph title: Western blot analysis ... Primary antibodies against the following proteins were used: Nox1 and Nox4 (Santa Cruz Biotechnology), superoxide dismutase (SOD) 1 and 2 (Abcam), heme oxygenase (HO)-1 (BD Biosciences, San Jose, CA, USA), NAD(P)H: quinone oxidoreductase-1 (NQO-1; Santa Cruz Biotechnology), catalase (Abcam), B-cell lymphoma 2 (Bcl-2; Santa Cruz Biotechnology), Kelch-like ECH-associated protein 1 (Keap1; Abcam), Nrf2 (Abcam), and p300/CBP-associated factor (PCAF; Santa Cruz Biotechnology).

Article Title: TCF7L1 indicates prognosis and promotes proliferation through activation of Keap1/NRF2 in gastric cancer
Article Snippet: Paragraph title: Western blot analysis ... Antibodies against TCF7L1 and NRF2 were purchased from Abcam (Cambridge, UK).

Article Title: Blockade of nicotine sensitization by methanol extracts of Glycyrrhizae radix mediated via antagonism of accumbal oxidative stress
Article Snippet: Paragraph title: Western blot analysis ... The primary antibodies for the immunoblotting assays included a rabbit polyclonal antibody for Nrf2 (Abcam, Cambridge, UK) and a rabbit polyclonal antibody for HO-1 (Abcam); the secondary antibody was IRDye 800CW Goat anti-Rabbit IgG (Li-Cor Bioscience, Lincoln, NE, USA).

Article Title: Oral Monomethyl Fumarate Therapy Ameliorates Retinopathy in a Humanized Mouse Model of Sickle Cell Disease
Article Snippet: Paragraph title: Western blot analysis ... Protein samples were subjected to SDS–PAGE, transferred to nitrocellulose membranes, and then incubated with primary antibodies: HbF (1:1000; Bethyl Laboratories, Inc.), Nrf2 (1:1000; AbCam), ZO-1 (1:100; Abcam), and VEGF (1:100; Abcam) overnight at 4°C.

Cell Culture:

Article Title: Tert-butylhydroquinone attenuates the ethanol-induced apoptosis of and activates the Nrf2 antioxidant defense pathway in H9c2 cardiomyocytes
Article Snippet: The H9c2 cells were seeded at 1.5×104 cells/well in glass chamber slides (glass coverslips) and cultured overnight at 37°C. .. The cells were then incubated overnight with primary antibodies against AC-tubulin (6-11B-1; ab24610) at 1:200 dilution and Nrf2 (both from Abcam) at a 1:200 dilution in a humidified chamber at 4°C.

Article Title: Brain ischemic preconditioning protects against ischemic injury and preserves the blood-brain barrier via oxidative signaling and Nrf2 activation
Article Snippet: PVDF membranes were then blocked with 5% non-fat milk, and probed with antibodies recognizing hemo oxygenase-1 (HO-1, 1:1000, Enzo Life Science, Farmingdale, NY), Nrf2 (1:1000, Abcam, Cambridge, MA), glutamate-cysteine ligase modifier subunit (GCLM, 1:1000, Abcam), β-actin (1:3000, Sigma-Aldrich, St. Louis, MO), FLAG (1:1000, Cell Signaling Technology, Danvers, MA), cadherin 5 (CDH5, 1:1000, Abcam), claudin 5 (1:1000, Abcam), Akt (1:1000, Cell Signaling Technology), phosphorylated Akt (p-Akt, S473, 1:1000, Cell Signaling Technology), GSK3β (1:1000, Cell Signaling Technology), phosphorylated GSK3β (p-GSK3β, S9, 1:1000, Cell Signaling Technology), human influenza hemagglutinin (HA, 1:1000, Cell Signaling Technology) or lamin B1 (1:1000, Cell Signaling Technology). .. For the Western dot blot assays, proteins were extracted from the brain tissues or cultured cells, and equivalent amounts of proteins were loaded directly onto PVDF membranes using a Bio-Dot Microfiltration apparatus (Bio-Rad, Hercules, CA).

Article Title: Cerebroprotection by salvianolic acid B after experimental subarachnoid hemorrhage occurs via Nrf2- and SIRT1-dependent pathways
Article Snippet: .. Briefly, brain sections (6 μm) or cultured cells were incubated with primary antibodies against 8-hydroxyguanosine (8-OHdG, Abcam), Nrf2 (1:100, Abcam), SIRT1 (1:50, Santa Cruz), caspase-3 (1:100, Cell Signaling), and NeuN (1:200, cat# MAB377, EMD Millipore, USA) overnight at 4 °C. .. Brain sections were then incubated with corresponding secondary antibodies Alexa Fluor 488 and/or Alexa Fluor 594 (Jackson ImmunoResearch Incorporation, West Grove, PA, USA).

Imaging:

Article Title: Carnosol as a Nrf2 Activator Improves Endothelial Barrier Function Through Antioxidative Mechanisms
Article Snippet: Assessment Expression of HO-1 and Nrf2 Proteins Antibodies for HO-1, β-actin were purchased from Proteintech (Rosemont, IL, USA), antibody for Nrf2 was purchased from Abcam (Cambridge, UK). .. The protein bands were detected by ECL reagents with the Bio-Rad ChemiDocTM MP Imaging System (Hercules, CA, USA).

Article Title: Blockade of nicotine sensitization by methanol extracts of Glycyrrhizae radix mediated via antagonism of accumbal oxidative stress
Article Snippet: The primary antibodies for the immunoblotting assays included a rabbit polyclonal antibody for Nrf2 (Abcam, Cambridge, UK) and a rabbit polyclonal antibody for HO-1 (Abcam); the secondary antibody was IRDye 800CW Goat anti-Rabbit IgG (Li-Cor Bioscience, Lincoln, NE, USA). .. The ODYSSEY Infrared Imaging System (Li-Cor Bioscience) was used to detect signals according to the manufacturer’s manual.

Inverted Microscopy:

Article Title: Cerebroprotection by salvianolic acid B after experimental subarachnoid hemorrhage occurs via Nrf2- and SIRT1-dependent pathways
Article Snippet: Briefly, brain sections (6 μm) or cultured cells were incubated with primary antibodies against 8-hydroxyguanosine (8-OHdG, Abcam), Nrf2 (1:100, Abcam), SIRT1 (1:50, Santa Cruz), caspase-3 (1:100, Cell Signaling), and NeuN (1:200, cat# MAB377, EMD Millipore, USA) overnight at 4 °C. .. Fluorescence was examined under a ZEISS HB050 inverted microscope system.

Protein Concentration:

Article Title: Apurinic/Apyrimidinic Endonuclease/Redox Factor-1 (APE1/Ref-1) Redox Function Negatively Regulates NRF2 *
Article Snippet: Whole cell lysates were prepared by lysing the cells in RIPA buffer (Santa Cruz Biotechnology, Santa Cruz, CA), followed by quantification of protein concentration (Lowry protein assay). .. Proteins were separated by SDS-PAGE, electroblotted onto nitrocellulose, and immunoblotting was performed using the following antibodies: Ref-1 and NRF2 (Abcam 62352), HMOX-1 (Abcam, Cambridge, MA), and Ku70, tubulin, or Actin (Sigma).

Article Title: Hormetic potential of methylglyoxal, a side-product of glycolysis, in switching tumours from growth to death
Article Snippet: Protein concentration is determined using bicinchoninic acid assay (Pierce, Carlsbad, CA). .. Blocking was performed in 5% non-fat dried milk (Biorad, Hercules, CA) in TBS-Tween 0.1% for 1 h. Membranes were then incubated overnight at 4 °C with primary antibodies: Argpyrimidine (1/10,000, mAb6B), MG-H1 (1/1000, STA-011, Cell Biolabs), Nrf2 (1/1000, Ab62353, Abcam), GLO1 (1/1000, #02-14, BioMac, Leipzig, Germany), β-actin (1/5000, A5441, Sigma).

Article Title: Oral Monomethyl Fumarate Therapy Ameliorates Retinopathy in a Humanized Mouse Model of Sickle Cell Disease
Article Snippet: Protein concentration was determined using the bicinchoninic acid assay (Thermo Fisher Scientific). .. Protein samples were subjected to SDS–PAGE, transferred to nitrocellulose membranes, and then incubated with primary antibodies: HbF (1:1000; Bethyl Laboratories, Inc.), Nrf2 (1:1000; AbCam), ZO-1 (1:100; Abcam), and VEGF (1:100; Abcam) overnight at 4°C.

Recombinant:

Article Title: Brg1-mediated Nrf2/HO-1 pathway activation alleviates hepatic ischemia–reperfusion injury
Article Snippet: Antibodies and reagents Antibodies recognizing Brg1 and Nrf2 were purchased from Abcam Company (Cambridge, MA, USA). .. Recombinant adenoviruses containing Brg1 (Brg1-Adv, 1 × 1010 pfu/ml) or HO-1(NM_010442, HO-1-Adv, 1 × 1010 pfu/ml) were designed and prepared from GeneCopoeia company.

Article Title: Ginkgetin induces autophagic cell death through p62/SQSTM1-mediated autolysosome formation and redox setting in non-small cell lung cancer
Article Snippet: The antibodies were obtained from the following sources: p62, TRAF6, and Nrf2 were obtained from Abcam (Cambridge, UK); α-tubulin from Sigma-Aldrich; and LC3 I/II, c-JUN, p-AKT, cleaved-PARP, cleaved-caspase 3, cleaved-caspase 7, ATG3, ATG5, ATG7, ATG12, ATG16L1, p-mTOR, mTOR, raptor, GβL, horseradish peroxidase (HRP)-conjugated goat anti-rabbit antibody, HRP-conjugated goat anti-mouse antibody, Alexa Fluor 555-conjugated goat anti-mouse antibody, and Alexa Fluor 488-conjugated goat anti-rabbit antibody from Cell Signaling Technology (Danvers, MA). p62 siRNA was obtained from Cell Signaling. .. The recombinant p62/SQSTM1 protein was purchased from Novus Biologicals (Littleton, CO).

Immunofluorescence:

Article Title: Tert-butylhydroquinone attenuates the ethanol-induced apoptosis of and activates the Nrf2 antioxidant defense pathway in H9c2 cardiomyocytes
Article Snippet: Paragraph title: Immunofluorescence staining ... The cells were then incubated overnight with primary antibodies against AC-tubulin (6-11B-1; ab24610) at 1:200 dilution and Nrf2 (both from Abcam) at a 1:200 dilution in a humidified chamber at 4°C.

Article Title: Cerebroprotection by salvianolic acid B after experimental subarachnoid hemorrhage occurs via Nrf2- and SIRT1-dependent pathways
Article Snippet: Paragraph title: 2.13. Immunofluorescence staining ... Briefly, brain sections (6 μm) or cultured cells were incubated with primary antibodies against 8-hydroxyguanosine (8-OHdG, Abcam), Nrf2 (1:100, Abcam), SIRT1 (1:50, Santa Cruz), caspase-3 (1:100, Cell Signaling), and NeuN (1:200, cat# MAB377, EMD Millipore, USA) overnight at 4 °C.

Article Title: Nrf2, a regulator of the proteasome, controls self-renewal and pluripotency in human embryonic stem cells
Article Snippet: Paragraph title: Immunofluorescence ... Samples were stained with primary antibodies Oct4 (Santa Cruz), Nanog (R & D), TuJ1 (Covance) and Nrf2 (Abcam) overnight at 4°C and secondary antibodies Alexa Fluor 488-donkey anti-mouse IgG, Alexa Fluor 488-donkey anti-rabbit IgG, Alexa Fluor 555-donkey anti-rabbit IgG, Alexa Fluor 488-donkey anti-goat IgG (Invitrogen).

Fluorescence:

Article Title: Tert-butylhydroquinone attenuates the ethanol-induced apoptosis of and activates the Nrf2 antioxidant defense pathway in H9c2 cardiomyocytes
Article Snippet: The cells were then incubated overnight with primary antibodies against AC-tubulin (6-11B-1; ab24610) at 1:200 dilution and Nrf2 (both from Abcam) at a 1:200 dilution in a humidified chamber at 4°C. .. After the slides were rinsed with PBS, coverslips were mounted on slides, and images of the labeled cells were visualized and photographed using a confocal fluorescence microscope (TCS SP2; Leica, Wetzlar, Germany).

Article Title: Cerebroprotection by salvianolic acid B after experimental subarachnoid hemorrhage occurs via Nrf2- and SIRT1-dependent pathways
Article Snippet: Briefly, brain sections (6 μm) or cultured cells were incubated with primary antibodies against 8-hydroxyguanosine (8-OHdG, Abcam), Nrf2 (1:100, Abcam), SIRT1 (1:50, Santa Cruz), caspase-3 (1:100, Cell Signaling), and NeuN (1:200, cat# MAB377, EMD Millipore, USA) overnight at 4 °C. .. Fluorescence was examined under a ZEISS HB050 inverted microscope system.

Isolation:

Article Title: Oral Monomethyl Fumarate Therapy Ameliorates Retinopathy in a Humanized Mouse Model of Sickle Cell Disease
Article Snippet: Protein was extracted from NR and RPE/eyecup isolated from control and MMF-treated HbAA- and HbSS-expressing mouse eyes. .. Protein samples were subjected to SDS–PAGE, transferred to nitrocellulose membranes, and then incubated with primary antibodies: HbF (1:1000; Bethyl Laboratories, Inc.), Nrf2 (1:1000; AbCam), ZO-1 (1:100; Abcam), and VEGF (1:100; Abcam) overnight at 4°C.

Microscopy:

Article Title: Tert-butylhydroquinone attenuates the ethanol-induced apoptosis of and activates the Nrf2 antioxidant defense pathway in H9c2 cardiomyocytes
Article Snippet: The cells were then incubated overnight with primary antibodies against AC-tubulin (6-11B-1; ab24610) at 1:200 dilution and Nrf2 (both from Abcam) at a 1:200 dilution in a humidified chamber at 4°C. .. After the slides were rinsed with PBS, coverslips were mounted on slides, and images of the labeled cells were visualized and photographed using a confocal fluorescence microscope (TCS SP2; Leica, Wetzlar, Germany).

Dot Blot:

Article Title: Brain ischemic preconditioning protects against ischemic injury and preserves the blood-brain barrier via oxidative signaling and Nrf2 activation
Article Snippet: PVDF membranes were then blocked with 5% non-fat milk, and probed with antibodies recognizing hemo oxygenase-1 (HO-1, 1:1000, Enzo Life Science, Farmingdale, NY), Nrf2 (1:1000, Abcam, Cambridge, MA), glutamate-cysteine ligase modifier subunit (GCLM, 1:1000, Abcam), β-actin (1:3000, Sigma-Aldrich, St. Louis, MO), FLAG (1:1000, Cell Signaling Technology, Danvers, MA), cadherin 5 (CDH5, 1:1000, Abcam), claudin 5 (1:1000, Abcam), Akt (1:1000, Cell Signaling Technology), phosphorylated Akt (p-Akt, S473, 1:1000, Cell Signaling Technology), GSK3β (1:1000, Cell Signaling Technology), phosphorylated GSK3β (p-GSK3β, S9, 1:1000, Cell Signaling Technology), human influenza hemagglutinin (HA, 1:1000, Cell Signaling Technology) or lamin B1 (1:1000, Cell Signaling Technology). .. For the Western dot blot assays, proteins were extracted from the brain tissues or cultured cells, and equivalent amounts of proteins were loaded directly onto PVDF membranes using a Bio-Dot Microfiltration apparatus (Bio-Rad, Hercules, CA).

Labeling:

Article Title: Tert-butylhydroquinone attenuates the ethanol-induced apoptosis of and activates the Nrf2 antioxidant defense pathway in H9c2 cardiomyocytes
Article Snippet: The cells were then incubated overnight with primary antibodies against AC-tubulin (6-11B-1; ab24610) at 1:200 dilution and Nrf2 (both from Abcam) at a 1:200 dilution in a humidified chamber at 4°C. .. After the slides were rinsed with PBS, coverslips were mounted on slides, and images of the labeled cells were visualized and photographed using a confocal fluorescence microscope (TCS SP2; Leica, Wetzlar, Germany).

Polyacrylamide Gel Electrophoresis:

Article Title: Blockade of nicotine sensitization by methanol extracts of Glycyrrhizae radix mediated via antagonism of accumbal oxidative stress
Article Snippet: Protein samples were separated using 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA). .. The primary antibodies for the immunoblotting assays included a rabbit polyclonal antibody for Nrf2 (Abcam, Cambridge, UK) and a rabbit polyclonal antibody for HO-1 (Abcam); the secondary antibody was IRDye 800CW Goat anti-Rabbit IgG (Li-Cor Bioscience, Lincoln, NE, USA).

Staining:

Article Title: Tert-butylhydroquinone attenuates the ethanol-induced apoptosis of and activates the Nrf2 antioxidant defense pathway in H9c2 cardiomyocytes
Article Snippet: Paragraph title: Immunofluorescence staining ... The cells were then incubated overnight with primary antibodies against AC-tubulin (6-11B-1; ab24610) at 1:200 dilution and Nrf2 (both from Abcam) at a 1:200 dilution in a humidified chamber at 4°C.

Article Title: Cerebroprotection by salvianolic acid B after experimental subarachnoid hemorrhage occurs via Nrf2- and SIRT1-dependent pathways
Article Snippet: Paragraph title: 2.13. Immunofluorescence staining ... Briefly, brain sections (6 μm) or cultured cells were incubated with primary antibodies against 8-hydroxyguanosine (8-OHdG, Abcam), Nrf2 (1:100, Abcam), SIRT1 (1:50, Santa Cruz), caspase-3 (1:100, Cell Signaling), and NeuN (1:200, cat# MAB377, EMD Millipore, USA) overnight at 4 °C.

Article Title: Nrf2, a regulator of the proteasome, controls self-renewal and pluripotency in human embryonic stem cells
Article Snippet: .. Samples were stained with primary antibodies Oct4 (Santa Cruz), Nanog (R & D), TuJ1 (Covance) and Nrf2 (Abcam) overnight at 4°C and secondary antibodies Alexa Fluor 488-donkey anti-mouse IgG, Alexa Fluor 488-donkey anti-rabbit IgG, Alexa Fluor 555-donkey anti-rabbit IgG, Alexa Fluor 488-donkey anti-goat IgG (Invitrogen). .. Cells were lysed in RIPA lysis buffer (Millipore) containing protease inhibitor cocktail (Roche).

SDS Page:

Article Title: Apurinic/Apyrimidinic Endonuclease/Redox Factor-1 (APE1/Ref-1) Redox Function Negatively Regulates NRF2 *
Article Snippet: .. Proteins were separated by SDS-PAGE, electroblotted onto nitrocellulose, and immunoblotting was performed using the following antibodies: Ref-1 and NRF2 (Abcam 62352), HMOX-1 (Abcam, Cambridge, MA), and Ku70, tubulin, or Actin (Sigma). .. The production of ROS was determined by detecting the fluorescent intensity of the oxidation-sensitive probe dihydrorhodamine 123 (DHR) (Molecular Probes, Invitrogen).

Article Title: KEAP1 Is a Redox Sensitive Target That Arbitrates the Opposing Radiosensitive Effects of Parthenolide in Normal and Cancer Cells
Article Snippet: .. Homogenized cells and tumor tissues were electrophoresed on an 8% (w/v) SDS-PAGE gel, transferred onto a nitrocellulose membrane, and subsequently incubated with primary antibodies against Keap1 (abcam), Nrf2 (abcam), MnSOD (Upstate Biotech), CuZnSOD (eBiosci), Gpx (abcam), catalase (Millipore), TrX (BD Sciences), PGAM5 (Santa Cruz Biotech.), Bcl-xL (Santa Cruz Biotech.), LC3B (cell signaling), β-actin (sigma), and pCNA (Santa Cruz Biotech.). ..

Article Title: Peroxiredoxin I is important for cancer-cell survival in Ras-induced hepatic tumorigenesis
Article Snippet: Protein samples were extracted by an SDS-PAGE and transferred to nitrocellulose membranes (Millipore, Bellerica, MA, USA). .. The membranes were primarily blotted with primary antibodies against Prx I, PCNA, Nrf2 (Abcam, Cambridge, UK); HA (Roche, Basel, Switzerland); GAPDH, FoxM1 (Abfrontier, Seoul, Korea); pERK, pP38, pJNK, cyclin D1 (Cell Signaling Technology, Danvers, MA, USA); and H-ras (Santa Cruz Biotechnology, Dallas, TX, USA).

Article Title: Hormetic potential of methylglyoxal, a side-product of glycolysis, in switching tumours from growth to death
Article Snippet: Twenty µg of protein were separated on 10 or 12.5% SDS-PAGE and transferred to PVDF membranes (Roche). .. Blocking was performed in 5% non-fat dried milk (Biorad, Hercules, CA) in TBS-Tween 0.1% for 1 h. Membranes were then incubated overnight at 4 °C with primary antibodies: Argpyrimidine (1/10,000, mAb6B), MG-H1 (1/1000, STA-011, Cell Biolabs), Nrf2 (1/1000, Ab62353, Abcam), GLO1 (1/1000, #02-14, BioMac, Leipzig, Germany), β-actin (1/5000, A5441, Sigma).

Article Title: T-type calcium channel blocker attenuates unilateral ureteral obstruction-induced renal interstitial fibrosis by activating the Nrf2 antioxidant pathway
Article Snippet: Equal amounts of proteins were separated by 15% SDS-PAGE and transferred to nitrocellulose membranes (Bio-Rad, Hercules, CA, USA). .. Primary antibodies against the following proteins were used: Nox1 and Nox4 (Santa Cruz Biotechnology), superoxide dismutase (SOD) 1 and 2 (Abcam), heme oxygenase (HO)-1 (BD Biosciences, San Jose, CA, USA), NAD(P)H: quinone oxidoreductase-1 (NQO-1; Santa Cruz Biotechnology), catalase (Abcam), B-cell lymphoma 2 (Bcl-2; Santa Cruz Biotechnology), Kelch-like ECH-associated protein 1 (Keap1; Abcam), Nrf2 (Abcam), and p300/CBP-associated factor (PCAF; Santa Cruz Biotechnology).

Article Title: TCF7L1 indicates prognosis and promotes proliferation through activation of Keap1/NRF2 in gastric cancer
Article Snippet: Equal amount of total proteins were subject to SDS-PAGE separation and then transferred to PVDF membranes (Millipore, Billerica, USA). .. Antibodies against TCF7L1 and NRF2 were purchased from Abcam (Cambridge, UK).

Article Title: Blockade of nicotine sensitization by methanol extracts of Glycyrrhizae radix mediated via antagonism of accumbal oxidative stress
Article Snippet: Protein samples were separated using 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA). .. The primary antibodies for the immunoblotting assays included a rabbit polyclonal antibody for Nrf2 (Abcam, Cambridge, UK) and a rabbit polyclonal antibody for HO-1 (Abcam); the secondary antibody was IRDye 800CW Goat anti-Rabbit IgG (Li-Cor Bioscience, Lincoln, NE, USA).

Article Title: Oral Monomethyl Fumarate Therapy Ameliorates Retinopathy in a Humanized Mouse Model of Sickle Cell Disease
Article Snippet: .. Protein samples were subjected to SDS–PAGE, transferred to nitrocellulose membranes, and then incubated with primary antibodies: HbF (1:1000; Bethyl Laboratories, Inc.), Nrf2 (1:1000; AbCam), ZO-1 (1:100; Abcam), and VEGF (1:100; Abcam) overnight at 4°C. .. Detection of HbF, Nrf2, and ZO-1 was done using horseradish peroxidase-conjugated secondary antibodies.

Plasmid Preparation:

Article Title: Ginkgetin induces autophagic cell death through p62/SQSTM1-mediated autolysosome formation and redox setting in non-small cell lung cancer
Article Snippet: The antibodies were obtained from the following sources: p62, TRAF6, and Nrf2 were obtained from Abcam (Cambridge, UK); α-tubulin from Sigma-Aldrich; and LC3 I/II, c-JUN, p-AKT, cleaved-PARP, cleaved-caspase 3, cleaved-caspase 7, ATG3, ATG5, ATG7, ATG12, ATG16L1, p-mTOR, mTOR, raptor, GβL, horseradish peroxidase (HRP)-conjugated goat anti-rabbit antibody, HRP-conjugated goat anti-mouse antibody, Alexa Fluor 555-conjugated goat anti-mouse antibody, and Alexa Fluor 488-conjugated goat anti-rabbit antibody from Cell Signaling Technology (Danvers, MA). p62 siRNA was obtained from Cell Signaling. .. The p62 plasmid was kindly gifted by Dr. Alexander Shneider from CureLab Oncology, Inc. (Needham, MA), and the pARE-Luc plasmid was obtained from Promega Corporation (Madison, WI).

Software:

Article Title: Brain ischemic preconditioning protects against ischemic injury and preserves the blood-brain barrier via oxidative signaling and Nrf2 activation
Article Snippet: PVDF membranes were then blocked with 5% non-fat milk, and probed with antibodies recognizing hemo oxygenase-1 (HO-1, 1:1000, Enzo Life Science, Farmingdale, NY), Nrf2 (1:1000, Abcam, Cambridge, MA), glutamate-cysteine ligase modifier subunit (GCLM, 1:1000, Abcam), β-actin (1:3000, Sigma-Aldrich, St. Louis, MO), FLAG (1:1000, Cell Signaling Technology, Danvers, MA), cadherin 5 (CDH5, 1:1000, Abcam), claudin 5 (1:1000, Abcam), Akt (1:1000, Cell Signaling Technology), phosphorylated Akt (p-Akt, S473, 1:1000, Cell Signaling Technology), GSK3β (1:1000, Cell Signaling Technology), phosphorylated GSK3β (p-GSK3β, S9, 1:1000, Cell Signaling Technology), human influenza hemagglutinin (HA, 1:1000, Cell Signaling Technology) or lamin B1 (1:1000, Cell Signaling Technology). .. ImageJ software was used for gel analyses.

Article Title: Hormetic potential of methylglyoxal, a side-product of glycolysis, in switching tumours from growth to death
Article Snippet: Blocking was performed in 5% non-fat dried milk (Biorad, Hercules, CA) in TBS-Tween 0.1% for 1 h. Membranes were then incubated overnight at 4 °C with primary antibodies: Argpyrimidine (1/10,000, mAb6B), MG-H1 (1/1000, STA-011, Cell Biolabs), Nrf2 (1/1000, Ab62353, Abcam), GLO1 (1/1000, #02-14, BioMac, Leipzig, Germany), β-actin (1/5000, A5441, Sigma). .. Bands quantification by densitometry and normalization to ß actin was performed using ImageJ software (NIH, Bethesda, MD).

Article Title: Cerebroprotection by salvianolic acid B after experimental subarachnoid hemorrhage occurs via Nrf2- and SIRT1-dependent pathways
Article Snippet: Briefly, brain sections (6 μm) or cultured cells were incubated with primary antibodies against 8-hydroxyguanosine (8-OHdG, Abcam), Nrf2 (1:100, Abcam), SIRT1 (1:50, Santa Cruz), caspase-3 (1:100, Cell Signaling), and NeuN (1:200, cat# MAB377, EMD Millipore, USA) overnight at 4 °C. .. The fluorescently stained cells were analyzed by Image J software.

Article Title: T-type calcium channel blocker attenuates unilateral ureteral obstruction-induced renal interstitial fibrosis by activating the Nrf2 antioxidant pathway
Article Snippet: Primary antibodies against the following proteins were used: Nox1 and Nox4 (Santa Cruz Biotechnology), superoxide dismutase (SOD) 1 and 2 (Abcam), heme oxygenase (HO)-1 (BD Biosciences, San Jose, CA, USA), NAD(P)H: quinone oxidoreductase-1 (NQO-1; Santa Cruz Biotechnology), catalase (Abcam), B-cell lymphoma 2 (Bcl-2; Santa Cruz Biotechnology), Kelch-like ECH-associated protein 1 (Keap1; Abcam), Nrf2 (Abcam), and p300/CBP-associated factor (PCAF; Santa Cruz Biotechnology). .. The density of the bands was analyzed using ImageJ software.

Binding Assay:

Article Title: T-type calcium channel blocker attenuates unilateral ureteral obstruction-induced renal interstitial fibrosis by activating the Nrf2 antioxidant pathway
Article Snippet: After electroblotting, nonspecific binding was blocked with 5% skim milk. .. Primary antibodies against the following proteins were used: Nox1 and Nox4 (Santa Cruz Biotechnology), superoxide dismutase (SOD) 1 and 2 (Abcam), heme oxygenase (HO)-1 (BD Biosciences, San Jose, CA, USA), NAD(P)H: quinone oxidoreductase-1 (NQO-1; Santa Cruz Biotechnology), catalase (Abcam), B-cell lymphoma 2 (Bcl-2; Santa Cruz Biotechnology), Kelch-like ECH-associated protein 1 (Keap1; Abcam), Nrf2 (Abcam), and p300/CBP-associated factor (PCAF; Santa Cruz Biotechnology).

Article Title: Oral Monomethyl Fumarate Therapy Ameliorates Retinopathy in a Humanized Mouse Model of Sickle Cell Disease
Article Snippet: Protein samples were subjected to SDS–PAGE, transferred to nitrocellulose membranes, and then incubated with primary antibodies: HbF (1:1000; Bethyl Laboratories, Inc.), Nrf2 (1:1000; AbCam), ZO-1 (1:100; Abcam), and VEGF (1:100; Abcam) overnight at 4°C. .. For the determination of VEGF protein levels, the retinal extracts were subjected to heparin binding affinity columns following previously described methods ( , ).

Next-Generation Sequencing:

Article Title: Tert-butylhydroquinone attenuates the ethanol-induced apoptosis of and activates the Nrf2 antioxidant defense pathway in H9c2 cardiomyocytes
Article Snippet: The cells were then permeabilized with 0.1% Triton X-100, washed and incubated with blocking buffer (10% NGS) for 1 h at room temperature. .. The cells were then incubated overnight with primary antibodies against AC-tubulin (6-11B-1; ab24610) at 1:200 dilution and Nrf2 (both from Abcam) at a 1:200 dilution in a humidified chamber at 4°C.

Acid Assay:

Article Title: Brain ischemic preconditioning protects against ischemic injury and preserves the blood-brain barrier via oxidative signaling and Nrf2 activation
Article Snippet: Protein concentrations were determined by the bicinchoninic acid assay (Bio-Rad, Hercules, CA). .. PVDF membranes were then blocked with 5% non-fat milk, and probed with antibodies recognizing hemo oxygenase-1 (HO-1, 1:1000, Enzo Life Science, Farmingdale, NY), Nrf2 (1:1000, Abcam, Cambridge, MA), glutamate-cysteine ligase modifier subunit (GCLM, 1:1000, Abcam), β-actin (1:3000, Sigma-Aldrich, St. Louis, MO), FLAG (1:1000, Cell Signaling Technology, Danvers, MA), cadherin 5 (CDH5, 1:1000, Abcam), claudin 5 (1:1000, Abcam), Akt (1:1000, Cell Signaling Technology), phosphorylated Akt (p-Akt, S473, 1:1000, Cell Signaling Technology), GSK3β (1:1000, Cell Signaling Technology), phosphorylated GSK3β (p-GSK3β, S9, 1:1000, Cell Signaling Technology), human influenza hemagglutinin (HA, 1:1000, Cell Signaling Technology) or lamin B1 (1:1000, Cell Signaling Technology).

Article Title: Hormetic potential of methylglyoxal, a side-product of glycolysis, in switching tumours from growth to death
Article Snippet: Protein concentration is determined using bicinchoninic acid assay (Pierce, Carlsbad, CA). .. Blocking was performed in 5% non-fat dried milk (Biorad, Hercules, CA) in TBS-Tween 0.1% for 1 h. Membranes were then incubated overnight at 4 °C with primary antibodies: Argpyrimidine (1/10,000, mAb6B), MG-H1 (1/1000, STA-011, Cell Biolabs), Nrf2 (1/1000, Ab62353, Abcam), GLO1 (1/1000, #02-14, BioMac, Leipzig, Germany), β-actin (1/5000, A5441, Sigma).

Article Title: Oral Monomethyl Fumarate Therapy Ameliorates Retinopathy in a Humanized Mouse Model of Sickle Cell Disease
Article Snippet: Protein concentration was determined using the bicinchoninic acid assay (Thermo Fisher Scientific). .. Protein samples were subjected to SDS–PAGE, transferred to nitrocellulose membranes, and then incubated with primary antibodies: HbF (1:1000; Bethyl Laboratories, Inc.), Nrf2 (1:1000; AbCam), ZO-1 (1:100; Abcam), and VEGF (1:100; Abcam) overnight at 4°C.

Immunoprecipitation:

Article Title: KEAP1 Is a Redox Sensitive Target That Arbitrates the Opposing Radiosensitive Effects of Parthenolide in Normal and Cancer Cells
Article Snippet: Paragraph title: Immunoblots and immunoprecipitation ... Homogenized cells and tumor tissues were electrophoresed on an 8% (w/v) SDS-PAGE gel, transferred onto a nitrocellulose membrane, and subsequently incubated with primary antibodies against Keap1 (abcam), Nrf2 (abcam), MnSOD (Upstate Biotech), CuZnSOD (eBiosci), Gpx (abcam), catalase (Millipore), TrX (BD Sciences), PGAM5 (Santa Cruz Biotech.), Bcl-xL (Santa Cruz Biotech.), LC3B (cell signaling), β-actin (sigma), and pCNA (Santa Cruz Biotech.).

Lysis:

Article Title: Peroxiredoxin I is important for cancer-cell survival in Ras-induced hepatic tumorigenesis
Article Snippet: Liver tissues and cell lysates were put in a lysis buffer (20 mM HEPES, 150 mL NaCl, 2 mM EGTA, 1 mM EDTA, 20 mM glycerol phosphate, 1% Triton X-100 and 10% glycerol) with protease (sigma, St Louise, MO) and a phosphatase-inhibitor cocktail (Roche, Basel, Switzerland). .. The membranes were primarily blotted with primary antibodies against Prx I, PCNA, Nrf2 (Abcam, Cambridge, UK); HA (Roche, Basel, Switzerland); GAPDH, FoxM1 (Abfrontier, Seoul, Korea); pERK, pP38, pJNK, cyclin D1 (Cell Signaling Technology, Danvers, MA, USA); and H-ras (Santa Cruz Biotechnology, Dallas, TX, USA).

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  • 90
    Abcam anti nrf2
    Decitabine and GO-203 combination decreases expression of DNMT1, 3b, increases Nox4, Duox2, activates <t>Nrf2</t> and Smad signaling pathway leading to downregulation of c-Myc A–B, H9 and HuT-78 cells were left untreated, treated with 3 uM GO-203 each day for 3 days, single dose of 40 nM Decitabine or the combination. Cells were harvested at 96 hours. Lysates were immunoblotted with the indicated antibodies.
    Anti Nrf2, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 120 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Decitabine and GO-203 combination decreases expression of DNMT1, 3b, increases Nox4, Duox2, activates Nrf2 and Smad signaling pathway leading to downregulation of c-Myc A–B, H9 and HuT-78 cells were left untreated, treated with 3 uM GO-203 each day for 3 days, single dose of 40 nM Decitabine or the combination. Cells were harvested at 96 hours. Lysates were immunoblotted with the indicated antibodies.

    Journal: Molecular cancer therapeutics

    Article Title: Decitabine Priming Enhances Mucin 1 Inhibition Mediated Disruption of Redox Homeostasis in Cutaneous T-cell Lymphoma

    doi: 10.1158/1535-7163.MCT-17-0060

    Figure Lengend Snippet: Decitabine and GO-203 combination decreases expression of DNMT1, 3b, increases Nox4, Duox2, activates Nrf2 and Smad signaling pathway leading to downregulation of c-Myc A–B, H9 and HuT-78 cells were left untreated, treated with 3 uM GO-203 each day for 3 days, single dose of 40 nM Decitabine or the combination. Cells were harvested at 96 hours. Lysates were immunoblotted with the indicated antibodies.

    Article Snippet: Soluble proteins were analyzed by immunoblotting with anti-DNMT1, 3b (Abcam), anti-Histone 3 (tri methyl K27) (Abcam), anti-TIGAR (Abcam), anti-phospho-p38 (Abcam), anti-Total p38 (Abcam), anti-phospho JNK (Abcam), anti-Total JNK (Abcam), anti-Nrf2 (Abcam), anti-phospho Smad2 and Smad3 (Cell-Signaling Technology), anti-Total Smad2 and Smad3 (Cell-Signaling Technology), anti-c-Myc (Abcam) and anti–GAPDH (Cell-Signaling Technology).

    Techniques: Expressing

    Postulated model of the signal transduction of VEGF 165 -induced Nrf2 activation. VEGF receptor ( VEGF-R ) activation-induced ERK1/2 phosphorylation leads to Nrf2 liberation from the Nrf2 inhibitor Keap1. In the nucleus, Nrf2 binds to the cis ) and via CO to promote VEGF expression by increasing hypoxia-inducible factor-1α ( HIF1 ). HRE , hypoxia response element.

    Journal: The Journal of Biological Chemistry

    Article Title: Interplay between Vascular Endothelial Growth Factor (VEGF) and Nuclear Factor Erythroid 2-related Factor-2 (Nrf2)

    doi: 10.1074/jbc.M111.286880

    Figure Lengend Snippet: Postulated model of the signal transduction of VEGF 165 -induced Nrf2 activation. VEGF receptor ( VEGF-R ) activation-induced ERK1/2 phosphorylation leads to Nrf2 liberation from the Nrf2 inhibitor Keap1. In the nucleus, Nrf2 binds to the cis ) and via CO to promote VEGF expression by increasing hypoxia-inducible factor-1α ( HIF1 ). HRE , hypoxia response element.

    Article Snippet: Antibodies against Nrf2, HO-1, thioredoxin, TXNRD1, and p38 were purchased from Abcam.

    Techniques: Transduction, Activation Assay, Expressing

    Nrf2 band migration rate on Bis-Tris gradient gels Cells were treated with 5 μM SFN for 2 h, and whole cell lysates were analyzed by 4–12% Bis-Tris SDS PAGE and western blotting. The same blot was probed with H-300 (left), EP1808Y (center), and C-20 (right) antibodies. Arrows indicate the positions at which Nrf2 migrates. Blots are representative of n = 2 biological replicates.

    Journal: Toxicology letters

    Article Title: Comparison of human Nrf2 antibodies: A tale of two proteins

    doi: 10.1016/j.toxlet.2015.07.004

    Figure Lengend Snippet: Nrf2 band migration rate on Bis-Tris gradient gels Cells were treated with 5 μM SFN for 2 h, and whole cell lysates were analyzed by 4–12% Bis-Tris SDS PAGE and western blotting. The same blot was probed with H-300 (left), EP1808Y (center), and C-20 (right) antibodies. Arrows indicate the positions at which Nrf2 migrates. Blots are representative of n = 2 biological replicates.

    Article Snippet: Here, an apparently highly specific Nrf2 monoclonal antibody, EP1808Y from Abcam, is compared with the most widely used Nrf2 antibodies, H-300 and C-20, both from Santa Cruz Biotechnology, in a panel of human cell lines.

    Techniques: Migration, SDS Page, Western Blot

    Comparison of EP1808Y monoclonal antibody with two commonly used Nrf2 polyclonal antibodies Cells were treated with 5 μM SFN for 2 h, and whole cell lysates were analyzed by 10% Tris-glycine SDS PAGE and western blotting. (A) Detection using the EP1808Y antibody at a brightness level that illustrates a single band is detected in HepG2 cell lysates and unchanged by SFN treatment. (B) The truncated blot in A is shown at full length (center), and brightness is increased to show detection in HaCaT and THLE-2 lysates. The same blot was probed with H-300 (left) and C-20 (right) antibodies. Arrows indicate the positions at which Nrf2 migrates. M, marker. Blots are representative of n = 3 biological replicates.

    Journal: Toxicology letters

    Article Title: Comparison of human Nrf2 antibodies: A tale of two proteins

    doi: 10.1016/j.toxlet.2015.07.004

    Figure Lengend Snippet: Comparison of EP1808Y monoclonal antibody with two commonly used Nrf2 polyclonal antibodies Cells were treated with 5 μM SFN for 2 h, and whole cell lysates were analyzed by 10% Tris-glycine SDS PAGE and western blotting. (A) Detection using the EP1808Y antibody at a brightness level that illustrates a single band is detected in HepG2 cell lysates and unchanged by SFN treatment. (B) The truncated blot in A is shown at full length (center), and brightness is increased to show detection in HaCaT and THLE-2 lysates. The same blot was probed with H-300 (left) and C-20 (right) antibodies. Arrows indicate the positions at which Nrf2 migrates. M, marker. Blots are representative of n = 3 biological replicates.

    Article Snippet: Here, an apparently highly specific Nrf2 monoclonal antibody, EP1808Y from Abcam, is compared with the most widely used Nrf2 antibodies, H-300 and C-20, both from Santa Cruz Biotechnology, in a panel of human cell lines.

    Techniques: SDS Page, Western Blot, Marker

    Immunodepletion and Nrf2 knockdown used to assess the primary band detected in HepG2 cells by EP1808Y (A) Schematic of the three Nrf2 isoforms from the three known alternatively-spliced transcripts with respect to H-300 and EP1808Y antibody epitopes. The transcript numbering aligns with that of the protein isoforms. Three commercially available siRNAs for Nrf2 mRNA target the indicated sites on the transcript. (B) HepG2 cells were treated with 5 μM SFN or 5 μM MG132 for 2 h, and whole cell lysates were immunodepleted with indicated antibodies, or beads alone (−). Depleted lysates were then analyzed by 10% Tris-glycine SDS PAGE and western blotting. The same blot was probed with H-300 and EP1808Y antibodies. Blots are representative of n = 3 biological replicates. (C) HepG2 cells were reverse transfected with siRNA (10 μM individual, 30 μM combined) for 24 h and then were treated with 5 μM SFN for 2 h. Whole cell lysates were analyzed by 10% Tris-glycine SDS PAGE and western blotting. The same blot was probed with H-300 and EP1808Y antibodies. C siRNA, control siRNA. Blot is representative of n = 2 biological replicates.

    Journal: Toxicology letters

    Article Title: Comparison of human Nrf2 antibodies: A tale of two proteins

    doi: 10.1016/j.toxlet.2015.07.004

    Figure Lengend Snippet: Immunodepletion and Nrf2 knockdown used to assess the primary band detected in HepG2 cells by EP1808Y (A) Schematic of the three Nrf2 isoforms from the three known alternatively-spliced transcripts with respect to H-300 and EP1808Y antibody epitopes. The transcript numbering aligns with that of the protein isoforms. Three commercially available siRNAs for Nrf2 mRNA target the indicated sites on the transcript. (B) HepG2 cells were treated with 5 μM SFN or 5 μM MG132 for 2 h, and whole cell lysates were immunodepleted with indicated antibodies, or beads alone (−). Depleted lysates were then analyzed by 10% Tris-glycine SDS PAGE and western blotting. The same blot was probed with H-300 and EP1808Y antibodies. Blots are representative of n = 3 biological replicates. (C) HepG2 cells were reverse transfected with siRNA (10 μM individual, 30 μM combined) for 24 h and then were treated with 5 μM SFN for 2 h. Whole cell lysates were analyzed by 10% Tris-glycine SDS PAGE and western blotting. The same blot was probed with H-300 and EP1808Y antibodies. C siRNA, control siRNA. Blot is representative of n = 2 biological replicates.

    Article Snippet: Here, an apparently highly specific Nrf2 monoclonal antibody, EP1808Y from Abcam, is compared with the most widely used Nrf2 antibodies, H-300 and C-20, both from Santa Cruz Biotechnology, in a panel of human cell lines.

    Techniques: SDS Page, Western Blot, Transfection

    Comparison of D1Z9C monoclonal antibody with H-300 in the panel of cell lines Cells were treated with SFN, as indicated for 2 h, and whole cell lysates were analyzed by 10% Tris-glycine SDS PAGE and western blotting. Arrows indicate the location of bands that correspond to Nrf2 as indicated by their increase in response to SFN. Blots are representative of n = 2 biological replicates.

    Journal: Toxicology letters

    Article Title: Comparison of human Nrf2 antibodies: A tale of two proteins

    doi: 10.1016/j.toxlet.2015.07.004

    Figure Lengend Snippet: Comparison of D1Z9C monoclonal antibody with H-300 in the panel of cell lines Cells were treated with SFN, as indicated for 2 h, and whole cell lysates were analyzed by 10% Tris-glycine SDS PAGE and western blotting. Arrows indicate the location of bands that correspond to Nrf2 as indicated by their increase in response to SFN. Blots are representative of n = 2 biological replicates.

    Article Snippet: Here, an apparently highly specific Nrf2 monoclonal antibody, EP1808Y from Abcam, is compared with the most widely used Nrf2 antibodies, H-300 and C-20, both from Santa Cruz Biotechnology, in a panel of human cell lines.

    Techniques: SDS Page, Western Blot

    MiR-144 mediates the increased Nrf2 transcriptional activity by Rg1 under OGD/R. a PC12 cells were treated with vehicle or Rg1 after 24 h followed with OGD, miRNA-144 level was determined by real-time PCR, and U6 was used as an internal control. Data were expressed as relative intensity of miR-144 normalized to U6 over control group. (** P

    Journal: Acta Pharmacologica Sinica

    Article Title: Ginsenoside Rg1 protects against ischemic/reperfusion-induced neuronal injury through miR-144/Nrf2/ARE pathway

    doi: 10.1038/s41401-018-0154-z

    Figure Lengend Snippet: MiR-144 mediates the increased Nrf2 transcriptional activity by Rg1 under OGD/R. a PC12 cells were treated with vehicle or Rg1 after 24 h followed with OGD, miRNA-144 level was determined by real-time PCR, and U6 was used as an internal control. Data were expressed as relative intensity of miR-144 normalized to U6 over control group. (** P

    Article Snippet: Primary antibodies against Nrf2, NQO-1, HO-1, glutamate cysteine ligase modifier subunit (GCLM), glutamate cysteine ligase catalytic subunit (GCLC), and Lamin A were purchased from Abcam (Cambridge, UK).

    Techniques: Activity Assay, Real-time Polymerase Chain Reaction

    Schematic diagram of the anti-I/R effect of Rg1 through miR-144/Nrf2/ARE pathway. I/R induces overproduction of ROS, which triggers oxidative stress and exhaust intracellular antioxidants. Meanwhile, it promotes antioxidants production by promoting the Nrf2/ARE pathway. I/R also promotes the activity of miR-144, which directly binds to Nrf2-3’-UTR in neurons and inhibits the expression of Nrf2. Rg1 inhibits the activity of miR-144 after I/R and promotes the Nrf2/ARE pathway, which enhances the expression of ARE-regulated antioxidative genes, reduces oxidative damage and exerts anti-I/R effect

    Journal: Acta Pharmacologica Sinica

    Article Title: Ginsenoside Rg1 protects against ischemic/reperfusion-induced neuronal injury through miR-144/Nrf2/ARE pathway

    doi: 10.1038/s41401-018-0154-z

    Figure Lengend Snippet: Schematic diagram of the anti-I/R effect of Rg1 through miR-144/Nrf2/ARE pathway. I/R induces overproduction of ROS, which triggers oxidative stress and exhaust intracellular antioxidants. Meanwhile, it promotes antioxidants production by promoting the Nrf2/ARE pathway. I/R also promotes the activity of miR-144, which directly binds to Nrf2-3’-UTR in neurons and inhibits the expression of Nrf2. Rg1 inhibits the activity of miR-144 after I/R and promotes the Nrf2/ARE pathway, which enhances the expression of ARE-regulated antioxidative genes, reduces oxidative damage and exerts anti-I/R effect

    Article Snippet: Primary antibodies against Nrf2, NQO-1, HO-1, glutamate cysteine ligase modifier subunit (GCLM), glutamate cysteine ligase catalytic subunit (GCLC), and Lamin A were purchased from Abcam (Cambridge, UK).

    Techniques: Activity Assay, Expressing

    Inhibition of miR-144 blocked the enhancement of Nrf2/ARE pathway induced by Rg1 after I/R. a Representative Western blot images of nuclear Nrf2 isolated from rat predicted penumbra cortex treated with vehicle or Rg1 (20 mg/kg) and transfected with control virus (NC) or miR-144-shRNA. Lamin A was used as an internal control. b Quantitative analysis of Nrf2 levels in a . (* P

    Journal: Acta Pharmacologica Sinica

    Article Title: Ginsenoside Rg1 protects against ischemic/reperfusion-induced neuronal injury through miR-144/Nrf2/ARE pathway

    doi: 10.1038/s41401-018-0154-z

    Figure Lengend Snippet: Inhibition of miR-144 blocked the enhancement of Nrf2/ARE pathway induced by Rg1 after I/R. a Representative Western blot images of nuclear Nrf2 isolated from rat predicted penumbra cortex treated with vehicle or Rg1 (20 mg/kg) and transfected with control virus (NC) or miR-144-shRNA. Lamin A was used as an internal control. b Quantitative analysis of Nrf2 levels in a . (* P

    Article Snippet: Primary antibodies against Nrf2, NQO-1, HO-1, glutamate cysteine ligase modifier subunit (GCLM), glutamate cysteine ligase catalytic subunit (GCLC), and Lamin A were purchased from Abcam (Cambridge, UK).

    Techniques: Inhibition, Western Blot, Isolation, Transfection, shRNA

    Rg1 promotes Nrf2 nuclear accumulation without change in Keap1. a Representative Western blot images of nuclear Nrf2 accumulation with different duration of OGD/R. Lamin A was used as an internal control. b Quantitative analysis of Nrf2 level in a (Normalized to 0 h for each group). * P

    Journal: Acta Pharmacologica Sinica

    Article Title: Ginsenoside Rg1 protects against ischemic/reperfusion-induced neuronal injury through miR-144/Nrf2/ARE pathway

    doi: 10.1038/s41401-018-0154-z

    Figure Lengend Snippet: Rg1 promotes Nrf2 nuclear accumulation without change in Keap1. a Representative Western blot images of nuclear Nrf2 accumulation with different duration of OGD/R. Lamin A was used as an internal control. b Quantitative analysis of Nrf2 level in a (Normalized to 0 h for each group). * P

    Article Snippet: Primary antibodies against Nrf2, NQO-1, HO-1, glutamate cysteine ligase modifier subunit (GCLM), glutamate cysteine ligase catalytic subunit (GCLC), and Lamin A were purchased from Abcam (Cambridge, UK).

    Techniques: Western Blot

    Rg1 improves OGD/R-induced neurotoxicity in an Nrf2-dependent manner in vitro. a Representative Western blot images for PC12 cells transfected with control-siRNA or Nrf2-siRNA, with or without Rg1 (1.00 μM) treatment. Lamin A was used as an internal control. b Quantitative analysis of Nrf2 level in a . (*** P

    Journal: Acta Pharmacologica Sinica

    Article Title: Ginsenoside Rg1 protects against ischemic/reperfusion-induced neuronal injury through miR-144/Nrf2/ARE pathway

    doi: 10.1038/s41401-018-0154-z

    Figure Lengend Snippet: Rg1 improves OGD/R-induced neurotoxicity in an Nrf2-dependent manner in vitro. a Representative Western blot images for PC12 cells transfected with control-siRNA or Nrf2-siRNA, with or without Rg1 (1.00 μM) treatment. Lamin A was used as an internal control. b Quantitative analysis of Nrf2 level in a . (*** P

    Article Snippet: Primary antibodies against Nrf2, NQO-1, HO-1, glutamate cysteine ligase modifier subunit (GCLM), glutamate cysteine ligase catalytic subunit (GCLC), and Lamin A were purchased from Abcam (Cambridge, UK).

    Techniques: In Vitro, Western Blot, Transfection