Nrf2 Antibody

Western Blot: Nrf2 Antibody [NBP3-09069] - Western blot analysis of Nrf2 on mouse liver tissue lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ER1706-41, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:5,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry/Immunofluorescence: Nrf2 Antibody [NBP3-09069] - ICC staining of Nrf2 in Huvec cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1706-41, 1/500) for 1 hour at room temperature, washed with PBS. Alexa Fluor (TM) 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

Immunohistochemistry-Paraffin: Nrf2 Antibody [NBP3-09069] - Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-Nrf2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Flow Cytometry: Nrf2 Antibody [NBP3-09069] - Flow cytometric analysis of Nrf2 was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (1/100) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Immunocytochemistry/Immunofluorescence: Nrf2 Antibody [NBP3-09069] - ICC staining of Nrf2 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1706-41, 1/500) for 1 hour at room temperature, washed with PBS. Alexa Fluor (TM) 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

Immunocytochemistry/Immunofluorescence: Nrf2 Antibody [NBP3-09069] - ICC staining of Nrf2 in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1706-41, 1/500) for 1 hour at room temperature, washed with PBS. Alexa Fluor (TM) 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

Immunohistochemistry-Paraffin: Nrf2 Antibody [NBP3-09069] - Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-Nrf2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemistry-Paraffin: Nrf2 Antibody [NBP3-09069] - Immunohistochemical analysis of paraffin-embedded human lung tissue using anti-Nrf2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemistry-Paraffin: Nrf2 Antibody [NBP3-09069] - Immunohistochemical analysis of paraffin-embedded mouse fallopian tubes tissue using anti-Nrf2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemistry-Paraffin: Nrf2 Antibody [NBP3-09069] - Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Nrf2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Product Discontinued

Nrf2 Antibody Summary

• Immunogen: Synthetic peptide within N-terminal human Nrf2. (SwissProt: Q16236 Human; SwissProt: Q60795 Mouse; SwissProt: O54968 Rat)
• Localization: Cytoplasm. Nucleus.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: NFE2L2
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:50-1:100
  • Immunocytochemistry/ Immunofluorescence 1:500-1:1000
  • Immunohistochemistry 1:50-1:200
  • Immunohistochemistry-Paraffin 1:50-1:200
  • Western Blot 1:500
• Theoretical MW: 68 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: TBS (pH7.4), 0.5% BSA, 50% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Nrf2 Antibody

• HEBP1
• NFE2L2
• NF-E2-related factor 2
• NFE2-related factor 2
• NRF2 erythroid derived 2, like 2
• Nrf2
• nuclear factor (erythroid-derived 2)-like 2
• nuclear factor erythroid 2-related factor 2
• nuclear factor erythroid-derived 2-like 2

Background

Nrf2 is a transcription factor which belongs to the Cap N Collar (CNC-bZIP) subfamily with an implication in adaptive responses to oxidative stress. Nrf2 is a basic leucine zipper transcription factor that can bind to a NF-E2/AP-1 repeat sequence in the promoter of the beta-globin gene and has two types of binding partners, a cytoplasmic repressor (Keap I) and multiple nuclear binding partners. During oxidative stress on a cell, Nrf2 is released from Keap I, possibly via PKC phosphorylation at serine 40, and translocates to the nucleus. In the nucleus, Nrf2 binds to antioxidant responsive element (ARE) and leads to transcriptional activation of downstream genes for GSTs, glutamate-cysteine ligase, HO-1, and thioredoxin. Nrf2 and its downstream proteins have shown to have important functions in protection against oxidative stress or chemical-induced cellular damage in liver and lung, prevention in cancer formation in GI tract, and promotion of the wound-healing process.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: NFE2L2