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Western Blot (WB) (Figure 1. Western blot analysis of NADPH oxidase 4/NOX4 using anti-NADPH oxidase 4/NOX4 antibody Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human HepG2 whole cell lysates, Lane 2: human SW620 whole cell lysates, Lane 3: human HK-2 whole cell lysates, Lane 4: human HL-60 whole cell lysates, Lane 5: human 293T whole cell lysates, Lane 6: human SW579 whole cell lysates, Lane 7: human SK-OV-3 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NADPH oxidase 4/NOX4 antigen affinity purified polyclonal antibody at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for NADPH OXIDASE 4/NOX4 at approximately 67KD. The expected band size for NADPH OXIDASE 4/NOX4 is at 67KD.)

Rabbit NADPH oxidase 4 Polyclonal Antibody | anti-NOX4 antibody

Anti-NADPH oxidase 4 antibody

Gene Names
Nox4; AI648021
Reactivity
Human, Rat; Predicted to work with: Mouse
Applications
Western Blot, Immunohistochemistry
Purity
Immunogen affinity purified.
Synonyms
NADPH oxidase 4; Polyclonal Antibody; Anti-NADPH oxidase 4 antibody; 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; anti-NOX4 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Rat; Predicted to work with: Mouse
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
No cross reactivity with other proteins
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
Sequence Length
578
Applicable Applications for anti-NOX4 antibody
Western Blot (WB), Immunohistochemistry (IHC) Paraffin
Application Notes
Western Blot:
Concentration:0.1-0.5ug/ml
Tested Species: Hu, Rat
Predicted Species: Ms

Immunohistochemistry (Paraffin-embedded Section):
Concentration:0.5-1ug/ml
Tested Species: Rat
Predicted Species: Hu,Ms
Antigen Retrieval: By Heat

Tested Species: In house tested species with positive results

Predicted Species: Species predicted to be fit for the product based on sequence similarities.

By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH 6.0, for 20mins is required for the staining of formalin/paraffin sections.

Other applications have not been tested.

MyBioSource provides a series of assays reacted with primary antibodies. Antibody can be supported by chemiluminescence kit MBS176460 in WB, supported by SA1022 in IHC(P), IHC(F) and ICC.
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of mouse NOX4(561-578aa NRNNSYGTKFEYNKES), identical to the related rat sequence and different from the related human sequence by two amino acids.
Reconstitution
0.2ml of distilled water will yield a concentration of 500ug/ml.
Preparation and Storage
At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. Western blot analysis of NADPH oxidase 4/NOX4 using anti-NADPH oxidase 4/NOX4 antibody Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human HepG2 whole cell lysates, Lane 2: human SW620 whole cell lysates, Lane 3: human HK-2 whole cell lysates, Lane 4: human HL-60 whole cell lysates, Lane 5: human 293T whole cell lysates, Lane 6: human SW579 whole cell lysates, Lane 7: human SK-OV-3 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NADPH oxidase 4/NOX4 antigen affinity purified polyclonal antibody at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for NADPH OXIDASE 4/NOX4 at approximately 67KD. The expected band size for NADPH OXIDASE 4/NOX4 is at 67KD.)

Western Blot (WB) (Figure 1. Western blot analysis of NADPH oxidase 4/NOX4 using anti-NADPH oxidase 4/NOX4 antibody Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human HepG2 whole cell lysates, Lane 2: human SW620 whole cell lysates, Lane 3: human HK-2 whole cell lysates, Lane 4: human HL-60 whole cell lysates, Lane 5: human 293T whole cell lysates, Lane 6: human SW579 whole cell lysates, Lane 7: human SK-OV-3 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NADPH oxidase 4/NOX4 antigen affinity purified polyclonal antibody at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for NADPH OXIDASE 4/NOX4 at approximately 67KD. The expected band size for NADPH OXIDASE 4/NOX4 is at 67KD.)

Immunohistochemistry (IHC)

(Figure 2. IHC analysis of NADPH OXIDASE 4/NOX4 using anti-NADPH OXIDASE 4/NOX4 antibody (PA1929). NADPH OXIDASE 4/NOX4 was detected in paraffin-embedded section of rat kidney tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-NADPH OXIDASE 4/NOX4 Antibody overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

Immunohistochemistry (IHC) (Figure 2. IHC analysis of NADPH OXIDASE 4/NOX4 using anti-NADPH OXIDASE 4/NOX4 antibody (PA1929). NADPH OXIDASE 4/NOX4 was detected in paraffin-embedded section of rat kidney tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-NADPH OXIDASE 4/NOX4 Antibody overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

Flow Cytometry (FC/FACS)

(Figure 3. Flow Cytometry analysis of U20S cells using anti-NADPH oxidase 4/NOX4 antibody Overlay histogram showing U20S cells stained (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti- NADPH oxidase 4/NOX4 Antibody (1ug/1x106 cells) for 30 min at 20 degree C. DyLight 488 conjugated goat anti-rabbit IgG (BA1127, 5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

Flow Cytometry (FC/FACS) (Figure 3. Flow Cytometry analysis of U20S cells using anti-NADPH oxidase 4/NOX4 antibody Overlay histogram showing U20S cells stained (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti- NADPH oxidase 4/NOX4 Antibody (1ug/1x106 cells) for 30 min at 20 degree C. DyLight 488 conjugated goat anti-rabbit IgG (BA1127, 5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

Western Blot (WB)

(Figure 4. Western blot analysis of NADPH oxidase 4/NOX4 using anti-NADPH oxidase 4/NOX4 antibody Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: rat kidney tissue lysate. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NADPH oxidase 4/NOX4 antigen affinity purified polyclonal antibody at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. Specific bands were detected for NADPH oxidase 4/NOX4 at approximately 67-80KD. The expected band size for NADPH oxidase 4/NOX4 are at 67-80KD.)

Western Blot (WB) (Figure 4. Western blot analysis of NADPH oxidase 4/NOX4 using anti-NADPH oxidase 4/NOX4 antibody Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: rat kidney tissue lysate. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NADPH oxidase 4/NOX4 antigen affinity purified polyclonal antibody at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. Specific bands were detected for NADPH oxidase 4/NOX4 at approximately 67-80KD. The expected band size for NADPH oxidase 4/NOX4 are at 67-80KD.)
Related Product Information for anti-NOX4 antibody
NADPH oxidase 4 is an enzyme that in humans is encoded by the NOX4 gene, and is a member of the NOX family of NADPH oxidases. This gene encodes a member of the NOX-family of enzymes that functions as the catalytic subunit the NADPH oxidase complex. The encoded protein is localized to non-phagocytic cells where it acts as an oxygen sensor and catalyzes the reduction of molecular oxygen to various reactive oxygen species (ROS). The ROS generated by this protein have been implicated in numerous biological functions including signal transduction, cell differentiation and tumor cell growth. A pseudogene has been identified on the other arm of chromosome 11. Alternative splicing results in multiple transcript variants

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
UniProt Accession #
NCBI Official Full Name
NADPH oxidase 4
NCBI Official Synonym Full Names
NADPH oxidase 4
NCBI Official Symbol
Nox4
NCBI Official Synonym Symbols
AI648021
NCBI Protein Information
NADPH oxidase 4; KOX-1; kidney oxidase-1; renal NAD(P)H-oxidase; superoxide-generating NADPH oxidase 4; kidney superoxide-producing NADPH oxidase
UniProt Protein Name
NADPH oxidase 4
Protein Family
UniProt Gene Name
Nox4
UniProt Synonym Gene Names
Renox; KOX-1
UniProt Entry Name
NOX4_MOUSE

Uniprot Description

NOX4: Constitutive NADPH oxidase which generates superoxide intracellularly upon formation of a complex with CYBA/p22phox. Regulates signaling cascades probably through phosphatases inhibition. May function as an oxygen sensor regulating the KCNK3/TASK-1 potassium channel and HIF1A activity. May regulate insulin signaling cascade. May play a role in apoptosis, bone resorption and lipolysaccharide-mediated activation of NFKB. May produce superoxide in the nucleus and play a role in regulating gene expression upon cell stimulation. Isoform 3 is not functional. Isoform 4 displays an increased activity. Isoform 5 and isoform 6 display reduced activity. 7 isoforms of the human protein are produced by alternative splicing.

Protein type: Oxidoreductase; Nucleolus; EC 1.6.3.-; Membrane protein, multi-pass; Membrane protein, integral; Cell cycle regulation

Cellular Component: focal adhesion; mitochondrion; membrane; endoplasmic reticulum; perinuclear region of cytoplasm; apical plasma membrane; integral to membrane; stress fiber; plasma membrane; cell junction; NADPH oxidase complex

Molecular Function: NAD(P)H oxidase activity; superoxide-generating NADPH oxidase activity; oxidoreductase activity, acting on NADH or NADPH, with oxygen as acceptor; oxidoreductase activity

Biological Process: positive regulation of apoptosis; cell morphogenesis; superoxide metabolic process; cell aging; homocysteine metabolic process; positive regulation of smooth muscle cell migration; cardiac muscle cell differentiation; positive regulation of protein kinase B signaling cascade; positive regulation of MAP kinase activity; negative regulation of cell proliferation; positive regulation of stress fiber formation; superoxide release; bone resorption

Research Articles on NOX4

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Product Notes

The NOX4 nox4 (Catalog #AAA176126) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-NADPH oxidase 4 antibody reacts with Human, Rat; Predicted to work with: Mouse and may cross-react with other species as described in the data sheet. AAA Biotech's NADPH oxidase 4 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC) Paraffin. Western Blot: Concentration:0.1-0.5ug/ml Tested Species: Hu, Rat Predicted Species: Ms Immunohistochemistry (Paraffin-embedded Section): Concentration:0.5-1ug/ml Tested Species: Rat Predicted Species: Hu,Ms Antigen Retrieval: By Heat Tested Species: In house tested species with positive results Predicted Species: Species predicted to be fit for the product based on sequence similarities. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH 6.0, for 20mins is required for the staining of formalin/paraffin sections. Other applications have not been tested. MyBioSource provides a series of assays reacted with primary antibodies. Antibody can be supported by chemiluminescence kit MBS176460 in WB, supported by SA1022 in IHC(P), IHC(F) and ICC. . Researchers should empirically determine the suitability of the NOX4 nox4 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "NADPH oxidase 4, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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