PRODUCT CODE: ET1607-4

NADPH oxidase 4/NOX4 Recombinant Rabbit Monoclonal Antibody [SY0214] (ET1607-4)

  • Recombinant

Applications

  • WB

  • ICC

  • IHC-P

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

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Western blot analysis of NADPH oxidase 4/NOX4 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 (ET1607-4, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: JAR cell lysate<br />
Lane 2: SH-SY5Y cell lysate
  • Western blot analysis of NADPH oxidase 4/NOX4 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 (ET1607-4, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: JAR cell lysate<br />
Lane 2: SH-SY5Y cell lysate
  • ICC staining of NADPH oxidase 4/NOX4 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 (ET1607-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of NADPH oxidase 4/NOX4 in A431 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 (ET1607-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of NADPH oxidase 4/NOX4 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 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1607-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human lung tissue using anti-NADPH oxidase 4/NOX4 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 (ET1607-4, 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.
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-NADPH oxidase 4/NOX4 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 (ET1607-4, 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.
  • Immunohistochemical analysis of paraffin-embedded rat stomach tissue using anti-NADPH oxidase 4/NOX4 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 (ET1607-4, 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.
  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-NADPH oxidase 4/NOX4 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 (ET1607-4, 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.
Western blot analysis of NADPH oxidase 4/NOX4 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 (ET1607-4, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: JAR cell lysate
Lane 2: SH-SY5Y cell lysate

Applications

  • WB

  • ICC

  • IHC-P

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

NADPH oxidase 4/NOX4 Recombinant Rabbit Monoclonal Antibody [SY0214] (ET1607-4)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

JAR cell lysate, SH-SY5Y cell lysate, Hela, A431, A549, human lung tissue, human kidney tissue, rat stomach tissue, rat kidney tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SY0214

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

67 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • ICC

  • 1:50-1:100

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

NADPH oxidase 4/NOX4

SYNONYMS

Kidney oxidase-1 antibody; Kidney superoxide-producing NADPH oxidase antibody; KOX 1 antibody; KOX antibody; Kox-1 antibody; NADPH antibody; NADPH oxidase 4 antibody; Nox4 antibody; NOX4_HUMAN antibody; Renal NAD(P)H-oxidase antibody; RENOX antibody

TISSUE SPECIFICITY

Expressed by distal tubular cells in kidney cortex and in endothelial cells (at protein level). Widely expressed. Strongly expressed in kidney and to a lower extent in heart, adipocytes, hepatoma, endothelial cells, skeletal muscle, brain, several brain tumor cell lines and airway epithelial cells.

DEVELOPMENTAL STAGE

Expressed in fetal kidney and fetal liver.

POST-TRANSLATIONAL MODIFICATION

Isoform 3 and isoform 4 are N-glycosylated. Isoform 4 glycosylation is required for its proper function.

SUBCELLULAR LOCATION

Cell membrane, Nucleus, Endoplasmic reticulum membrane.

FUNCTION

The superoxide-generating NADPH oxidase includes a membrane-bound flavocytochrome containing two subunits, gp91-phox and p22-phox, and the cytosolic proteins p47-phox and p67-phox. During activation of the NADPH oxidase, p47-phox and p67-phox migrate to the plasma membrane where they associate with the flavocytochrome, cytochrome b558, to form the active enzyme complex. The p22 and gp91-phox subunits also function as surface O2 sensors that initiate cellular signaling in response to hypoxic conditions. Nox4 (also known as Renox) is a renal gp91-phox homolog highly expressed at the site of erythropoietin production in the proximal convoluted tubule epithelial cells of the renal cortex. Nox4 is also expressed in fetal tissues, placenta, glioblastoma and vascular cells. Like gp91-phox, the enzymatic activity of Nox4 produces superoxide anions. In vascular cells, the addition of Angiotensin II increases Nox4 expression, which suggests a role for Nox4 in vascular oxidative stress response. The gene encoding human Nox4 maps to chromosome 11q14.2-q21.