Anti-Phospho-Nrf2(S40) antibody [SU0334]
cat.: ET1608-28
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, ICC/IF, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: SU0334
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1 mg/mL.
Purification: Protein A affinity purified.
Molecular weight: 90 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Ser40 of Human Nrf2 aa 20-69 / 605.
Positive control: HepG2 cell lysate, Raji cell lysate, Hela, A549, HepG2, human breast carcinoma tissue, human tonsil tissue, human kidney tissue, K562.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  ICC/IF
  IHC-P
  FC
  IP

1:1,000-1:5,000
1:50-1:200
1:50-1:200
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: Q16236 Human
Alternative names: erythroid derived 2 HEBP1 like 2 NF E2 related factor 2 NF-E2-related factor 2 NF2L2_HUMAN NFE2 related factor 2 NFE2-related factor 2 Nfe2l2 Nrf 2 NRF2 Nuclear factor (erythroid derived 2) like 2 Nuclear factor nuclear factor erythroid 2 like 2 Nuclear factor erythroid 2 related factor 2 Nuclear factor erythroid 2-related factor 2 Nuclear factor erythroid derived 2 like 2
Images
ET1608-28_1.jpg Fig1: Western blot analysis of Phospho-Nrf2(S40) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-28, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: HepG2 cell lysate
Lane 2: Raji cell lysate
ET1608-28_2.jpg Fig2: ICC staining of Phospho-Nrf2(S40) 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 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-28, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-28_3.jpg Fig3: ICC staining of Phospho-Nrf2(S40) in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-28, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-28_4.jpg Fig4: ICC staining of Phospho-Nrf2(S40) 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 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-28, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-28_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Phospho-Nrf2(S40) antibody [SU0334] (ET1608-28). The section was pre-treated using heat mediated antigen retrieval with 10 mM sodium citrate buffer (pH 6.0). The section were untreated (A, C) and treated (B) with λ-PPase at 1/25 dilution at 30℃ for 30 minutes after antigen retrieval. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-28, 1/200) for 30 minutes at room temperature. Goat anti-Rabbit IgG-HRP UltraPolymer antibody (HA1119) was used for 20 minutes at room temperature. DAB was used as the chromogen. The section were counterstained with hematoxylin and mounted with DPX. PBS was used instead of the primary antibody as the negative control, and is shown in the inset (C).
ET1608-28_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Phospho-Nrf2(S40) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-28, 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.
ET1608-28_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Phospho-Nrf2(S40) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-28, 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.
ET1608-28_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Phospho-Nrf2(S40) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-28, 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.
ET1608-28_9.jpg Fig9: Flow cytometric analysis of Phospho-Nrf2(S40) was done on K562 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1608-28, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
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