Structured Review

Santa Cruz Biotechnology anti keap1 antibody
Structure of the interface between <t>KEAP1</t> and Nrf2 and location of Cys434 . A bottom view ( left panel ) and a side view ( right panel ) of the KEAP1-DC structure as revealed by x-ray crystallography are shown. Cys434 is pink . Residues in KEAP1 involved in direct interaction with the ETGE motif are indicated: Arg380 , Arg415 , and Arg483 in dark blue ; Ser363 , Ser508 , Ser555 , and Ser602 in orange ; and Asn382 in light blue. Numbers of the blade structure ( 1–6 ) in KEAP1 are shown around the bottom view of the molecular model.
Anti Keap1 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 80 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti keap1 antibody - by Bioz Stars, 2020-01
80/100 stars

Images

1) Product Images from "The Critical Role of Nitric Oxide Signaling, via Protein "

Article Title: The Critical Role of Nitric Oxide Signaling, via Protein

Journal:

doi: 10.1074/jbc.M110.145441

Structure of the interface between KEAP1 and Nrf2 and location of Cys434 . A bottom view ( left panel ) and a side view ( right panel ) of the KEAP1-DC structure as revealed by x-ray crystallography are shown. Cys434 is pink . Residues in KEAP1 involved in direct interaction with the ETGE motif are indicated: Arg380 , Arg415 , and Arg483 in dark blue ; Ser363 , Ser508 , Ser555 , and Ser602 in orange ; and Asn382 in light blue. Numbers of the blade structure ( 1–6 ) in KEAP1 are shown around the bottom view of the molecular model.
Figure Legend Snippet: Structure of the interface between KEAP1 and Nrf2 and location of Cys434 . A bottom view ( left panel ) and a side view ( right panel ) of the KEAP1-DC structure as revealed by x-ray crystallography are shown. Cys434 is pink . Residues in KEAP1 involved in direct interaction with the ETGE motif are indicated: Arg380 , Arg415 , and Arg483 in dark blue ; Ser363 , Ser508 , Ser555 , and Ser602 in orange ; and Asn382 in light blue. Numbers of the blade structure ( 1–6 ) in KEAP1 are shown around the bottom view of the molecular model.

Techniques Used:

2) Product Images from "A novel tag-free probe for targeting molecules interacting with a flavonoid catabolite"

Article Title: A novel tag-free probe for targeting molecules interacting with a flavonoid catabolite

Journal: Biochemistry and Biophysics Reports

doi: 10.1016/j.bbrep.2016.06.020

Identification of the target proteins of DOPAC. Detection of the DPE-modified Keap1 and AhR. Confluent Hepa1c1c7 cells were incubated with 50 µM DPE for 5 h in serum-free MEM-α. The cell lysate was incubated with Streptavidin Mag Sepharose beads for 30 min. The DPE-modified Keap1 (left) or AhR (right) were detected by immunoblot analysis.
Figure Legend Snippet: Identification of the target proteins of DOPAC. Detection of the DPE-modified Keap1 and AhR. Confluent Hepa1c1c7 cells were incubated with 50 µM DPE for 5 h in serum-free MEM-α. The cell lysate was incubated with Streptavidin Mag Sepharose beads for 30 min. The DPE-modified Keap1 (left) or AhR (right) were detected by immunoblot analysis.

Techniques Used: Modification, Incubation

Related Articles

MTT Assay:

Article Title: The Activation of Nrf2 and Its Downstream Regulated Genes Mediates the Antioxidative Activities of Xueshuan Xinmaining Tablet in Human Umbilical Vein Endothelial Cells
Article Snippet: TACS MTT Cell Proliferation Assay Kit and GPX enzyme-linked immunosorbent assay (ELISA) were purchased from R & D Systems (Minneapolis, MN, USA). .. Anti-Nrf2, Keap1, GCLM, NQO1, HMOX1, and anti-Keap1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Protease Inhibitor:

Article Title: A novel tag-free probe for targeting molecules interacting with a flavonoid catabolite
Article Snippet: 2.1 DOPAC, laccase from Rhus vernicifera , glyceraldehyde-3-phosphate dehydrogenase (GAPDH), tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine (TBTA), bis(4-nitrophenyl) phosphate (BNPP) and azide-PEG3 -biotin conjugate were obtained from Sigma Aldrich (St. Louis, USA). p -Toluensulfonic acid monohydrate (PTSA), n -butanol, toluene, copper (II) sulfate pentahydrate, protease inhibitor cocktail and Chemi-Lumi One Super were purchased from nacalai tasque (Kyoto, Japan). .. Anti-actin antibody, anti-aryl hydrocarbon receptor (AhR) antibody, anti-Keap1 antibody, horseradish peroxidase-linked anti-mouse IgG and horseradish peroxidase-linked anti-goat IgG were purchased from Santa Cruz Biotechnology (Santa Cruz, USA).

Enzyme-linked Immunosorbent Assay:

Article Title: The Activation of Nrf2 and Its Downstream Regulated Genes Mediates the Antioxidative Activities of Xueshuan Xinmaining Tablet in Human Umbilical Vein Endothelial Cells
Article Snippet: TACS MTT Cell Proliferation Assay Kit and GPX enzyme-linked immunosorbent assay (ELISA) were purchased from R & D Systems (Minneapolis, MN, USA). .. Anti-Nrf2, Keap1, GCLM, NQO1, HMOX1, and anti-Keap1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Immunoprecipitation:

Article Title: S-Nitrosoglutathione Reductase Deficiency-Induced S-Nitrosylation Results in Neuromuscular Dysfunction
Article Snippet: Paragraph title: Immunoprecipitation assays ... Immunoprecipitations were performed by adding 1 μg of anti-Keap1 antibody (Santa Cruz) to 50 μl of prewashed Dynabeads® -protein G (Invitrogen) to whole tissue extracts from skeletal muscle.

Incubation:

Article Title: The Critical Role of Nitric Oxide Signaling, via Protein
Article Snippet: After membranes were blocked with TTBS (20 m m Tris-HCl, 150 m m NaCl, 0.1% Tween 20, pH 7.6) containing 5% skim milk (Difco), they were incubated with antibodies in TTBS containing 5% skim milk at 4 °C overnight. .. Other antibodies used in the Western blotting analysis were as follows: anti-Keap1 antibody (rat monoclonal) , anti-actin antibody (C-11, Santa Cruz Biotechnology, Inc. (Santa Cruz, CA)), anti-iNOS antibody (Santa Cruz Biotechnology, Inc.), anti-FLAG M2 antibody (Sigma), anti-HO-1 antibody (Stressgen Bioreagents, Victoria, Canada), and anti-NQO1 (NAD(P)H dehydrogenase, quinone 1) antibody (Santa Cruz Biotechnology, Inc.).

Article Title: S-Nitrosoglutathione Reductase Deficiency-Induced S-Nitrosylation Results in Neuromuscular Dysfunction
Article Snippet: Immunoprecipitations were performed by adding 1 μg of anti-Keap1 antibody (Santa Cruz) to 50 μl of prewashed Dynabeads® -protein G (Invitrogen) to whole tissue extracts from skeletal muscle. .. Immunoprecipitations were performed by adding 1 μg of anti-Keap1 antibody (Santa Cruz) to 50 μl of prewashed Dynabeads® -protein G (Invitrogen) to whole tissue extracts from skeletal muscle.

Proliferation Assay:

Article Title: The Activation of Nrf2 and Its Downstream Regulated Genes Mediates the Antioxidative Activities of Xueshuan Xinmaining Tablet in Human Umbilical Vein Endothelial Cells
Article Snippet: TACS MTT Cell Proliferation Assay Kit and GPX enzyme-linked immunosorbent assay (ELISA) were purchased from R & D Systems (Minneapolis, MN, USA). .. Anti-Nrf2, Keap1, GCLM, NQO1, HMOX1, and anti-Keap1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Expressing:

Article Title: The Critical Role of Nitric Oxide Signaling, via Protein
Article Snippet: Other antibodies used in the Western blotting analysis were as follows: anti-Keap1 antibody (rat monoclonal) , anti-actin antibody (C-11, Santa Cruz Biotechnology, Inc. (Santa Cruz, CA)), anti-iNOS antibody (Santa Cruz Biotechnology, Inc.), anti-FLAG M2 antibody (Sigma), anti-HO-1 antibody (Stressgen Bioreagents, Victoria, Canada), and anti-NQO1 (NAD(P)H dehydrogenase, quinone 1) antibody (Santa Cruz Biotechnology, Inc.). .. Other antibodies used in the Western blotting analysis were as follows: anti-Keap1 antibody (rat monoclonal) , anti-actin antibody (C-11, Santa Cruz Biotechnology, Inc. (Santa Cruz, CA)), anti-iNOS antibody (Santa Cruz Biotechnology, Inc.), anti-FLAG M2 antibody (Sigma), anti-HO-1 antibody (Stressgen Bioreagents, Victoria, Canada), and anti-NQO1 (NAD(P)H dehydrogenase, quinone 1) antibody (Santa Cruz Biotechnology, Inc.).

Polymerase Chain Reaction:

Article Title: The Activation of Nrf2 and Its Downstream Regulated Genes Mediates the Antioxidative Activities of Xueshuan Xinmaining Tablet in Human Umbilical Vein Endothelial Cells
Article Snippet: Human Oxidative Stress PCR Array was purchased from Qiagen (Valencia, CA, USA). .. Anti-Nrf2, Keap1, GCLM, NQO1, HMOX1, and anti-Keap1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Western Blot:

Article Title: The Critical Role of Nitric Oxide Signaling, via Protein
Article Snippet: For analysis of protein S -guanylation, we used two rabbit anti- S -guanylated protein antibodies ( i.e. 8-RS-cGMP and 8-RS-Guo antibodies) with different specificities and one monoclonal anti- S -guanylated (8-RS-cGMP) antibody, as described previously ( ) and in the . .. Other antibodies used in the Western blotting analysis were as follows: anti-Keap1 antibody (rat monoclonal) , anti-actin antibody (C-11, Santa Cruz Biotechnology, Inc. (Santa Cruz, CA)), anti-iNOS antibody (Santa Cruz Biotechnology, Inc.), anti-FLAG M2 antibody (Sigma), anti-HO-1 antibody (Stressgen Bioreagents, Victoria, Canada), and anti-NQO1 (NAD(P)H dehydrogenase, quinone 1) antibody (Santa Cruz Biotechnology, Inc.). .. Membranes were washed in TTBS three times and incubated with a horseradish peroxidase-conjugated secondary antibody at room temperature for 1 h. After three washes in TTBS, immunoreactive bands were detected by using a chemiluminescence reagent (ECL Plus Western blotting reagent; GE Healthcare) with a luminescent image analyzer (LAS-1000UVmini, Fujifilm (Tokyo, Japan)).

Article Title: Disturbance of redox status enhances radiosensitivity of hepatocellular carcinoma
Article Snippet: Paragraph title: Western blots ... The membrane was blocked for 1 h in TBST containing 0.5% FBS and subsequently probed with anti-Nrf2 antibody (Santa Cruz, CA, USA), anti-Keap1 antibody (Santa Cruz), anti-HO1 antibody (Cell Signaling, Danvers, USA) and anti-NQO1 antibody (Cell Signaling) at 4°C overnight with shaking.

SDS Page:

Article Title: The Critical Role of Nitric Oxide Signaling, via Protein
Article Snippet: Proteins in cell lysates were heat-denatured and separated via SDS-PAGE under reducing conditions and were transferred to polyvinylidene difluoride membranes (Immobilon-P, Millipore (Bedford, MA)). .. Other antibodies used in the Western blotting analysis were as follows: anti-Keap1 antibody (rat monoclonal) , anti-actin antibody (C-11, Santa Cruz Biotechnology, Inc. (Santa Cruz, CA)), anti-iNOS antibody (Santa Cruz Biotechnology, Inc.), anti-FLAG M2 antibody (Sigma), anti-HO-1 antibody (Stressgen Bioreagents, Victoria, Canada), and anti-NQO1 (NAD(P)H dehydrogenase, quinone 1) antibody (Santa Cruz Biotechnology, Inc.).

Article Title: Disturbance of redox status enhances radiosensitivity of hepatocellular carcinoma
Article Snippet: Proteins were separated by 10% SDS-PAGE and transferred to a methanol activated PVDF membrane. .. The membrane was blocked for 1 h in TBST containing 0.5% FBS and subsequently probed with anti-Nrf2 antibody (Santa Cruz, CA, USA), anti-Keap1 antibody (Santa Cruz), anti-HO1 antibody (Cell Signaling, Danvers, USA) and anti-NQO1 antibody (Cell Signaling) at 4°C overnight with shaking.

Article Title: S-Nitrosoglutathione Reductase Deficiency-Induced S-Nitrosylation Results in Neuromuscular Dysfunction
Article Snippet: Immunoprecipitations were performed by adding 1 μg of anti-Keap1 antibody (Santa Cruz) to 50 μl of prewashed Dynabeads® -protein G (Invitrogen) to whole tissue extracts from skeletal muscle. .. Immunoprecipitations were performed by adding 1 μg of anti-Keap1 antibody (Santa Cruz) to 50 μl of prewashed Dynabeads® -protein G (Invitrogen) to whole tissue extracts from skeletal muscle.

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  • 74
    Santa Cruz Biotechnology antibody against keap1
    ERK5 controls <t>Keap1</t> mRNA expression. A) 10 7 Jurkat-TAg cells were transfected with 5 μg of the empty pcDNA vector, ERK5 or a pSUPER Neo vector containing a small hairpin RNA for ERK5 (shERK5). Forty-eight hours later mRNA expression was analyzed by qPCR and presented as the % of mRNA compared to cells transfected with the control vector. B) Protein expression of cells transfected in (A). C) 10 7 Jurkat-TAg cells were transfected with 5 μg of the empty pSUPER Neo vector or with the vector encoding for the shERK5. Protein expression was analyzed by WB at different times after transfection. The data represent means ± SD; *p
    Antibody Against Keap1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 74/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibody against keap1/product/Santa Cruz Biotechnology
    Average 74 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    antibody against keap1 - by Bioz Stars, 2020-01
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    78
    Santa Cruz Biotechnology goat anti keap1 polyclonal antibody
    Increased mRNA expression of Nrf2-dependent genes and decreased <t>Keap1</t> transcript and protein levels in the lungs from Keap1Δ2–3/Δ2–3 ;CctCre+ mice. ( A ) Messenger RNA levels of Nqo1 and Gclm in the lungs of Keap1flox/flox mice and Keap1Δ2–3/Δ2–3 ;CctCre+ mice. Lungs were isolated from 8-week-old mice. The fold change was calculated using the formula described in M aterials and M ethods ( n = 3). ( B ) Messenger RNA levels of Keap1 in the lungs of Keap1flox/flox mice and Keap1Δ2–3/Δ2–3 ;CctCre+ mice. Data represented are mean fold change ± SEM ( n = 3). Asterisks indicate a significant difference compared with Keap1flox/flox mice ( P < 0.05). ( C ) Lung lysates from Keap1flox/flox and Keap1Δ2–3/Δ2–3 ;CctCre+ mice, and cell lysates from human cancer cells (positive control). Tissue and cell lysates were prepared and analyzed through immunoblotting using an anti-Keap1 goat <t>polyclonal</t> that detects the full-length protein. The blot was stripped and probed with an anti-GAPDH antibody to ensure equal protein loading between samples ( n = 1). ( D ) Densitometric analysis of Keap1 protein expression normalized to GAPDH protein expression for experiment shown in C . Protein band intensities were quantified using ImageJ software (National Institutes of Health, Bethesda, MD).
    Goat Anti Keap1 Polyclonal Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti keap1 polyclonal antibody/product/Santa Cruz Biotechnology
    Average 78 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti keap1 polyclonal antibody - by Bioz Stars, 2020-01
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    99
    Santa Cruz Biotechnology anti nrf2 antibody
    Proposed <t>Nrf2-ARE</t> signaling pathway. Nrf2 is expressed constitutively in the cell and translocates directly to the nucleus following its synthesis. Following transactivation of its genes, Nrf2 is targeted for degradation by Keap1 in the nucleus, a
    Anti Nrf2 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti nrf2 antibody/product/Santa Cruz Biotechnology
    Average 99 stars, based on 28 article reviews
    Price from $9.99 to $1999.99
    anti nrf2 antibody - by Bioz Stars, 2020-01
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    ERK5 controls Keap1 mRNA expression. A) 10 7 Jurkat-TAg cells were transfected with 5 μg of the empty pcDNA vector, ERK5 or a pSUPER Neo vector containing a small hairpin RNA for ERK5 (shERK5). Forty-eight hours later mRNA expression was analyzed by qPCR and presented as the % of mRNA compared to cells transfected with the control vector. B) Protein expression of cells transfected in (A). C) 10 7 Jurkat-TAg cells were transfected with 5 μg of the empty pSUPER Neo vector or with the vector encoding for the shERK5. Protein expression was analyzed by WB at different times after transfection. The data represent means ± SD; *p

    Journal: EBioMedicine

    Article Title: Human Leukemic Cells performing Oxidative Phosphorylation (OXPHOS) Generate an Antioxidant Response Independently of Reactive Oxygen species (ROS) Production

    doi: 10.1016/j.ebiom.2015.11.045

    Figure Lengend Snippet: ERK5 controls Keap1 mRNA expression. A) 10 7 Jurkat-TAg cells were transfected with 5 μg of the empty pcDNA vector, ERK5 or a pSUPER Neo vector containing a small hairpin RNA for ERK5 (shERK5). Forty-eight hours later mRNA expression was analyzed by qPCR and presented as the % of mRNA compared to cells transfected with the control vector. B) Protein expression of cells transfected in (A). C) 10 7 Jurkat-TAg cells were transfected with 5 μg of the empty pSUPER Neo vector or with the vector encoding for the shERK5. Protein expression was analyzed by WB at different times after transfection. The data represent means ± SD; *p

    Article Snippet: The antibody against KEAP1 and MEF2 (E-17) were from Santa Cruz Biotechnology.

    Techniques: Expressing, Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Western Blot

    Cells performing OXPHOS activate an antioxidant response. A) Different cell lines were grown in OXPHOS medium for at least 1 month before mRNA extraction. mRNA expression was quantified by qPCR and represented as the % of mRNA compared to control cells. B) Cells were treated with 20 mM DCA for 24 and 48 h and KEAP1 and NQO1 mRNA levels were quantified by qPCR. C) The expression of different proteins was analyzed in cells growing in OXPHOS medium or treated with DCA as described above. The data represent means ± SD; *p

    Journal: EBioMedicine

    Article Title: Human Leukemic Cells performing Oxidative Phosphorylation (OXPHOS) Generate an Antioxidant Response Independently of Reactive Oxygen species (ROS) Production

    doi: 10.1016/j.ebiom.2015.11.045

    Figure Lengend Snippet: Cells performing OXPHOS activate an antioxidant response. A) Different cell lines were grown in OXPHOS medium for at least 1 month before mRNA extraction. mRNA expression was quantified by qPCR and represented as the % of mRNA compared to control cells. B) Cells were treated with 20 mM DCA for 24 and 48 h and KEAP1 and NQO1 mRNA levels were quantified by qPCR. C) The expression of different proteins was analyzed in cells growing in OXPHOS medium or treated with DCA as described above. The data represent means ± SD; *p

    Article Snippet: The antibody against KEAP1 and MEF2 (E-17) were from Santa Cruz Biotechnology.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Increase in ROS levels is not essential for KEAP1 downregulation. A) Jurkat cells were treated with increasing concentrations of H 2 O 2 for 1 h and mRNA expression was analyzed. B) OCI-AML3 cells (left) or primary tumor cells from a BCL patient (right) were treated with 1.5 mM NAC 1 h before adding DCA (20 mM) for 24 h. Cells were labeled with CH-H2DCFDA and analyzed by FACs for ROS production. Keap1 mRNA and protein were analyzed as described in Fig. 2 . C) Primary tumor cells from 2 BCL patients were treated as in (B) before analyzing KEAP1 mRNA expression, results represent the means ± SD of these two patients in triplicate. The data represent means ± SD; *p

    Journal: EBioMedicine

    Article Title: Human Leukemic Cells performing Oxidative Phosphorylation (OXPHOS) Generate an Antioxidant Response Independently of Reactive Oxygen species (ROS) Production

    doi: 10.1016/j.ebiom.2015.11.045

    Figure Lengend Snippet: Increase in ROS levels is not essential for KEAP1 downregulation. A) Jurkat cells were treated with increasing concentrations of H 2 O 2 for 1 h and mRNA expression was analyzed. B) OCI-AML3 cells (left) or primary tumor cells from a BCL patient (right) were treated with 1.5 mM NAC 1 h before adding DCA (20 mM) for 24 h. Cells were labeled with CH-H2DCFDA and analyzed by FACs for ROS production. Keap1 mRNA and protein were analyzed as described in Fig. 2 . C) Primary tumor cells from 2 BCL patients were treated as in (B) before analyzing KEAP1 mRNA expression, results represent the means ± SD of these two patients in triplicate. The data represent means ± SD; *p

    Article Snippet: The antibody against KEAP1 and MEF2 (E-17) were from Santa Cruz Biotechnology.

    Techniques: Expressing, Labeling, FACS

    miR-23a targets KEAP1 mRNA. A) Jurkat cells were transfected with the whole miR-23a–27a–24-2 locus or with the constructs miR-23 ∆ 24–27 and miR-23 ∆ 23. The expression of KEAP1 mRNA was analyzed by qPCR and represented as the % of mRNA compared to cells transfected with the control vector. B) Expression of KEAP1 protein and the quantification. C) Jurkat cells were transfected with the different constructs together with a reporter plasmid containing the 3′UTR of KEAP1 mRNA downstream of the luciferase mRNA. Data are represented as the % of luciferase expression in cells transfected with the empty vector. D) The expression of NQO-1 mRNA was analyzed by qPCR in cells transfected as in (A). The data represent means ± SD; *p

    Journal: EBioMedicine

    Article Title: Human Leukemic Cells performing Oxidative Phosphorylation (OXPHOS) Generate an Antioxidant Response Independently of Reactive Oxygen species (ROS) Production

    doi: 10.1016/j.ebiom.2015.11.045

    Figure Lengend Snippet: miR-23a targets KEAP1 mRNA. A) Jurkat cells were transfected with the whole miR-23a–27a–24-2 locus or with the constructs miR-23 ∆ 24–27 and miR-23 ∆ 23. The expression of KEAP1 mRNA was analyzed by qPCR and represented as the % of mRNA compared to cells transfected with the control vector. B) Expression of KEAP1 protein and the quantification. C) Jurkat cells were transfected with the different constructs together with a reporter plasmid containing the 3′UTR of KEAP1 mRNA downstream of the luciferase mRNA. Data are represented as the % of luciferase expression in cells transfected with the empty vector. D) The expression of NQO-1 mRNA was analyzed by qPCR in cells transfected as in (A). The data represent means ± SD; *p

    Article Snippet: The antibody against KEAP1 and MEF2 (E-17) were from Santa Cruz Biotechnology.

    Techniques: Transfection, Construct, Expressing, Real-time Polymerase Chain Reaction, Plasmid Preparation, Luciferase

    Effects of XXT on Nrf2 and Keap1 mRNA expression levels in HUVECs.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: The Activation of Nrf2 and Its Downstream Regulated Genes Mediates the Antioxidative Activities of Xueshuan Xinmaining Tablet in Human Umbilical Vein Endothelial Cells

    doi: 10.1155/2015/187265

    Figure Lengend Snippet: Effects of XXT on Nrf2 and Keap1 mRNA expression levels in HUVECs.

    Article Snippet: Anti-Nrf2, Keap1, GCLM, NQO1, HMOX1, and anti-Keap1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Expressing

    Effects of XXT on the protein expression levels of Keap1, Nrf2, HMOX1, GCLM, and NQO1 in HUVECs.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: The Activation of Nrf2 and Its Downstream Regulated Genes Mediates the Antioxidative Activities of Xueshuan Xinmaining Tablet in Human Umbilical Vein Endothelial Cells

    doi: 10.1155/2015/187265

    Figure Lengend Snippet: Effects of XXT on the protein expression levels of Keap1, Nrf2, HMOX1, GCLM, and NQO1 in HUVECs.

    Article Snippet: Anti-Nrf2, Keap1, GCLM, NQO1, HMOX1, and anti-Keap1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Expressing

    Schematic representation of XXT activities on Keap1-Nrf2-ARE pathway.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: The Activation of Nrf2 and Its Downstream Regulated Genes Mediates the Antioxidative Activities of Xueshuan Xinmaining Tablet in Human Umbilical Vein Endothelial Cells

    doi: 10.1155/2015/187265

    Figure Lengend Snippet: Schematic representation of XXT activities on Keap1-Nrf2-ARE pathway.

    Article Snippet: Anti-Nrf2, Keap1, GCLM, NQO1, HMOX1, and anti-Keap1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques:

    Increased mRNA expression of Nrf2-dependent genes and decreased Keap1 transcript and protein levels in the lungs from Keap1Δ2–3/Δ2–3 ;CctCre+ mice. ( A ) Messenger RNA levels of Nqo1 and Gclm in the lungs of Keap1flox/flox mice and Keap1Δ2–3/Δ2–3 ;CctCre+ mice. Lungs were isolated from 8-week-old mice. The fold change was calculated using the formula described in M aterials and M ethods ( n = 3). ( B ) Messenger RNA levels of Keap1 in the lungs of Keap1flox/flox mice and Keap1Δ2–3/Δ2–3 ;CctCre+ mice. Data represented are mean fold change ± SEM ( n = 3). Asterisks indicate a significant difference compared with Keap1flox/flox mice ( P < 0.05). ( C ) Lung lysates from Keap1flox/flox and Keap1Δ2–3/Δ2–3 ;CctCre+ mice, and cell lysates from human cancer cells (positive control). Tissue and cell lysates were prepared and analyzed through immunoblotting using an anti-Keap1 goat polyclonal that detects the full-length protein. The blot was stripped and probed with an anti-GAPDH antibody to ensure equal protein loading between samples ( n = 1). ( D ) Densitometric analysis of Keap1 protein expression normalized to GAPDH protein expression for experiment shown in C . Protein band intensities were quantified using ImageJ software (National Institutes of Health, Bethesda, MD).

    Journal:

    Article Title: Deletion of Keap1 in the Lung Attenuates Acute Cigarette Smoke-Induced Oxidative Stress and Inflammation

    doi: 10.1165/rcmb.2009-0054OC

    Figure Lengend Snippet: Increased mRNA expression of Nrf2-dependent genes and decreased Keap1 transcript and protein levels in the lungs from Keap1Δ2–3/Δ2–3 ;CctCre+ mice. ( A ) Messenger RNA levels of Nqo1 and Gclm in the lungs of Keap1flox/flox mice and Keap1Δ2–3/Δ2–3 ;CctCre+ mice. Lungs were isolated from 8-week-old mice. The fold change was calculated using the formula described in M aterials and M ethods ( n = 3). ( B ) Messenger RNA levels of Keap1 in the lungs of Keap1flox/flox mice and Keap1Δ2–3/Δ2–3 ;CctCre+ mice. Data represented are mean fold change ± SEM ( n = 3). Asterisks indicate a significant difference compared with Keap1flox/flox mice ( P < 0.05). ( C ) Lung lysates from Keap1flox/flox and Keap1Δ2–3/Δ2–3 ;CctCre+ mice, and cell lysates from human cancer cells (positive control). Tissue and cell lysates were prepared and analyzed through immunoblotting using an anti-Keap1 goat polyclonal that detects the full-length protein. The blot was stripped and probed with an anti-GAPDH antibody to ensure equal protein loading between samples ( n = 1). ( D ) Densitometric analysis of Keap1 protein expression normalized to GAPDH protein expression for experiment shown in C . Protein band intensities were quantified using ImageJ software (National Institutes of Health, Bethesda, MD).

    Article Snippet: Membranes were exposed to either a goat anti-Keap1 polyclonal antibody (Santa Cruz Biotechnology, Santa Cruz, CA), a rabbit anti-Keap1 polyclonal antibody (Santa Cruz) in PBST-0.5% milk overnight at 4°C followed by incubation with horseradish peroxidase–conjugated secondary antibodies (GE Healthcare UK Ltd, Buckinghamshire, UK).

    Techniques: Expressing, Mouse Assay, Isolation, Positive Control, Software

    Proposed Nrf2-ARE signaling pathway. Nrf2 is expressed constitutively in the cell and translocates directly to the nucleus following its synthesis. Following transactivation of its genes, Nrf2 is targeted for degradation by Keap1 in the nucleus, a

    Journal:

    Article Title: The Nrf2-Antioxidant Response Element Signaling Pathway and Its Activation by Oxidative Stress

    doi: 10.1074/jbc.R900010200

    Figure Lengend Snippet: Proposed Nrf2-ARE signaling pathway. Nrf2 is expressed constitutively in the cell and translocates directly to the nucleus following its synthesis. Following transactivation of its genes, Nrf2 is targeted for degradation by Keap1 in the nucleus, a

    Article Snippet: Nrf2 was stained with an anti-Nrf2 antibody ( An important question is how Keap1 targets Nrf2 for ubiquitylation, given their localization in distinct cellular compartments.

    Techniques:

    Regulation of rat GSTA2 and NQO1 gene expression. Induction of these two detoxication enzymes is regulated at the transcriptional level mediated by two distinct enhancers, XRE and ARE, controlled by the AhR and Nrf2, respectively. β-Naphthoflavone

    Journal:

    Article Title: The Nrf2-Antioxidant Response Element Signaling Pathway and Its Activation by Oxidative Stress

    doi: 10.1074/jbc.R900010200

    Figure Lengend Snippet: Regulation of rat GSTA2 and NQO1 gene expression. Induction of these two detoxication enzymes is regulated at the transcriptional level mediated by two distinct enhancers, XRE and ARE, controlled by the AhR and Nrf2, respectively. β-Naphthoflavone

    Article Snippet: Nrf2 was stained with an anti-Nrf2 antibody ( An important question is how Keap1 targets Nrf2 for ubiquitylation, given their localization in distinct cellular compartments.

    Techniques: Expressing

    Nuclear localization of Nrf2 in human umbilical vein endothelial cells. ) and visualized with a secondary antibody conjugated

    Journal:

    Article Title: The Nrf2-Antioxidant Response Element Signaling Pathway and Its Activation by Oxidative Stress

    doi: 10.1074/jbc.R900010200

    Figure Lengend Snippet: Nuclear localization of Nrf2 in human umbilical vein endothelial cells. ) and visualized with a secondary antibody conjugated

    Article Snippet: Nrf2 was stained with an anti-Nrf2 antibody ( An important question is how Keap1 targets Nrf2 for ubiquitylation, given their localization in distinct cellular compartments.

    Techniques: