Nox4 Antibody

Images

 
Western Blot: Nox4 Antibody [NB110-58849] - Silencing of NOX4 in H4-II-C3 rat hepatoma cell line. Image from verified customer review.
Immunocytochemistry/ Immunofluorescence: Nox4 Antibody [NB110-58849] - Porcine aortic endothelial cells stained with NB110-58849 (green). Nuclei were counterstained with DAPI (blue). Image from verified customer review.
Immunohistochemistry: Nox4 Antibody [NB110-58849] - Detection of NOX4 in proximal convoluted tubules of the kidney using NB110-58849 at 5 ug/mL.
Western Blot: Nox4 Antibody [NB110-58849] - Detection of NOX4 in HUVEC whole cell lysate. Lanes 1 and 2: serum-starved HUVEC lysate denatured at 95C for 5 minutes. Lanes 3: serum-starved HUVEC lysate denatured at room ...read more
Biological Strategies: Western Blot: Nox4 Antibody [NB110-58849] - Detection of NOX4 on IMR90 lysate. Image courtesy of Naomi Logsdon, University of Alabama at Birmingham.
Western Blot: Nox4 Antibody [NB110-58849] - Detection of NOX4 in human kidney lysates using NB110-58849 at 2.0 ug/mL. A non-specific band is often observed running at 50 kDa in tissue lysates which is believed to ...read more
Immunocytochemistry/ Immunofluorescence: Nox4 Antibody [NB110-58849] - Detection of NOX4 in mouse brain showing positive staining in neurons in the cortex. Image provided by Rachel Reith.
Immunocytochemistry/ Immunofluorescence: Nox4 Antibody [NB110-58849] - NOX4 antibody was tested in Hek293 cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Bv, Pm, Rb, ShSpecies Glossary
Applications WB, IB, ICC/IF, IHC, IHC-Fr, IHC-P
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml
Validated by:
   

Biological Strategies

     

Nox4 Antibody Summary

Immunogen
A synthetic peptide made to an internal region of the human NOX4 protein (between residues 100-200) [UniProt Q9NPH5].
Localization
Endoplasmic reticulum, endoplasmic reticulum membrane, multi-pass membrane protein, cell membrane, nucleus (probable).
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
NOX4
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 2 ug/mL
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:50 - 1:200
  • Immunohistochemistry 5 ug/mL
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 5 ug/mL
Application Notes
Immunohistochemistry-Frozen was reported in scientific literature. In Western blot this antibody recognizes a band at ~67 kDa or larger representing isoform 1 of NOX4, and what appears to be a non-specific band ~48 kDa. The observed band size may vary depending on sample type and glycosylation. In ICC/IF cytoplasmic and mitochondrial staining is observed. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. Use in Immunoblotting reported in scientific literature (PMID 28152080).
Control Peptide
Nox4 Peptide (NB110-58849PEP)
Reviewed Applications
Read 7 Reviews rated 4.4
using
NB110-58849 in the following applications:

Publications
Read Publications using
NB110-58849 in the following applications:

Reactivity Notes

Porcine reactivity reported in scientific literature (PMID: 24403605)

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
PBS
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for Nox4 Antibody

  • EC 1.6.3
  • Kidney oxidase-1
  • Kidney superoxide-producing NADPH oxidase
  • KOX
  • KOX-1
  • KOX-1Renal NAD(P)H-oxidase
  • NADPH oxidase 4
  • Nox4
  • RENOX
  • RENOXEC 1.6.3.-

Background

NOX4 (NADPH oxidase 4) is a constitutive NADPH oxidase which complex with CYBA/p22phox for generating superoxide intracellularly and is found primarily in vascular cells, fibroblasts, osteoclasts and is abundantly expressed in the kidney. NOX4 regulates signaling cascades through phosphatases inhibition and function as oxygen sensor regulating the KCNK3/TASK-1 potassium channel as well as HIF1 alpha activity. NOX4 has also been implicated in regulation of insulin signaling cascade, apoptosis, bone resorption and LPS-mediated activation of NFkB. In vascular smooth muscle cells, NOX4 is positively regulated by Poldip2 which is a Nox4/p22phox interacting protein and Nox4/p22phox-Poldip2 complex regulates Rho-dependent cytoskeletal reorganization as well as cellular migration. TGF- beta is another positive regulator of Nox4 which is important in vascular fibrosis and differentiation. Unlike Nox1/Nox2, Nox4 is constitutively active, producing primarily H2O2 rather thanO2 and this difference in the species generated underlie NOX-specific actions in cell signaling. Nox4 contributes to basal ROS production through its constitutive activity and to increased ROS generation when stimulated by Ang II, glucose, TNF alpha and growth factors. NOX4 has been implicated in hypertension, atherosclerosis, and cardiovascular and renal complications of diabetes and in remodeling of pulmonary arteries in hypertension.

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.

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Publications for Nox4 Antibody (NB110-58849)(53)

We have publications tested in 6 confirmed species: Human, Mouse, Rat, Bovine, Porcine, Rabbit.

We have publications tested in 6 applications: IB, ICC/IF, IHC, IHC-Fr, IHC-P, WB.


Filter By Application
IB
(1)
ICC/IF
(6)
IHC
(4)
IHC-Fr
(1)
IHC-P
(2)
WB
(36)
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Filter By Species
Human
(16)
Mouse
(22)
Rat
(6)
Bovine
(1)
Porcine
(1)
Rabbit
(1)
All Species
Showing Publications 1 - 10 of 53. Show All 53 Publications.
Publications using NB110-58849 Applications Species
Liu R, Yan X. Sulforaphane protects rabbit corneas against oxidative stress injury in keratoconus through activation of the Nrf-2/HO-1 antioxidant pathway Int. J. Mol. Med. Aug 10 2018 [PMID: 30106111] (IHC-P, Rabbit) IHC-P Rabbit
Dai Y, Zhang H, Zhang J, Yan M. Isoquercetin attenuates oxidative stress and neuronal apoptosis after ischemia/reperfusion injury via Nrf2-mediated inhibition of the NOX4/ROS/NF-kB pathway. Chem. Biol. Interact. 2018 Feb 16 [PMID: 29454613] (WB, Rat) WB Rat
Moloney JN, Jayavelu AK, Stanicka J et al. Nuclear membrane-localised NOX4D generates pro-survival ROS in FLT3-ITD-expressing AML Oncotarget [PMID: 29285262] (WB, Human) WB Human
Shanmugasundaram K, Nayak BK, Friedrichs WE et al. NOX4 functions as a mitochondrial energetic sensor coupling cancer metabolic reprogramming to drug resistance. Nat Commun. 2017 Oct 19 [PMID: 29051480] (WB, Human) WB Human
Tsubouchi K, Araya J, Minagawa S et al. Azithromycin attenuates myofibroblast differentiation and lung fibrosis development through proteasomal degradation of NOX4. Autophagy. 2017 Jun 14 [PMID: 28613983] (ICC/IF, Human) ICC/IF Human
Wang L, Ma R, Guo Y et al. Antioxidant Effect of Fructus Ligustri Lucidi Aqueous Extract in Ovariectomized Rats Is Mediated through Nox4-ROS-NF-kB Pathway. Front Pharmacol. 2017 Jun 07 [PMID: 28588482] (IHC, WB, Rat) IHC, WB Rat
Kim YM, Kim SJ, Tatsunami R et al. ROS-induced ROS Release Orchestrated by Nox4, Nox2 and Mitochondria in VEGF Signaling and Angiogenesis. Am. J. Physiol., Cell Physiol. Apr 19 2017 [PMID: 28424170] (WB, Human) WB Human
Sigala F, Efentakis P, Karageorgiadi D et al. Reciprocal regulation of eNOS, H2S and CO-synthesizing enzymes in human atheroma: Correlation with plaque stability and effects of simvastatin. Redox Biol. 2017 Feb 12 [PMID: 28214453] (IHC, Human) IHC Human
Dickson BJ, Gatie MI, Spice DM, Kelly GM. NOX1 and NOX4 are required for the differentiation of mouse F9 cells into extraembryonic endoderm. PLoS ONE. 2017 Feb 02 [PMID: 28152080] (IB, Mouse) IB Mouse
Chen L, Zhao M, Li J et al. Critical role of X-box binding protein 1 in NADPH oxidase 4-triggered cardiac hypertrophy is mediated by receptor interacting protein kinase 1. Cell Cycle. Dec 8 2016 [PMID: 27929749]
Show All 53 Publications.

Reviews for Nox4 Antibody (NB110-58849) (7) 4.47

Average Rating: 4.4
(Based on 7 reviews)
We have 7 reviews tested in 4 species: Human, Mouse, Rat, Other.

Reviews using NB110-58849:
Filter by Applications
WB
(5)
IF
(1)
IHC-P
(1)
All Applications
Filter by Species
Human
(2)
Mouse
(2)
Rat
(1)
Other
(2)
All Species
Images Ratings Applications Species Date Details
Western Blot Nox4 NB110-58849
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reviewed by:
Anonymous
WB Rat 03/01/2018
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Summary

ApplicationWestern Blot
Sample TestedH4-II-E-C3 rat hepatoma cell line
SpeciesRat
Lotn
Immunofluorescence Nox4 NB110-58849
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reviewed by:
Anonymous
IF Other 07/25/2016
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Summary

ApplicationImmunofluorescence
Sample Testedporcine aortic endothelial cells
SpeciesOther
LotL

Blocking

Blocking Detailsblocking: PBS 2% BSA 1h at room temperature

Primary Anitbody

Dilution Ratio1:100 in PBS 1% BSA, 2h at 37°C

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG, alexa 488 conjugate, 1h at room temperature, in PBS 1% BSA
Secondary Manufacturer Cat#sc-362257
Secondary Concentration1:500

Details

Detection Notesepifluorescence, Leica DM2500 fluorescence microscope with a setting of 40X magnification, 200ms
Fixation Detailscells fixed in PFA 4% 15' at room temperature; permeabilization: 3x5' pbs-triton 0,1%
Wash Description3x5' PBS 0,5% BSA
Western Blot Nox4 NB110-58849
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reviewed by:
Anonymous
WB Other 07/22/2016
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Summary

ApplicationWestern Blot
Sample TestedPAOEC cell lysate
SpeciesOther
LotL

Blocking

Blocking Detailsmilk 5% in tbs 500 mM NaCl-tween 0,1%, 1h at room temp,

Primary Anitbody

Dilution Ratio1:1000 in milk 3% tbs 500 mM NaCl-tween 0,1%, overnight at 4°C

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG, HRP, in TBST 3% milk, 1h at room temperature
Secondary Manufacturer Cat#A0545 sigma-aldrich
Secondary Concentration1:3000

Details

Detection Noteswith Pierce ECL western blotting substrate, (2' incubation), and developed on Amersham Hyperfilm ECL, GE Healthcare, 2' exposure
 
reviewed by:
Paramananda Saikia
WB Mouse 07/01/2014
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Summary

ApplicationWestern Blot
Sample TestedMouse
SpeciesMouse
LotD

Blocking

Blocking Details5'sa

Primary Anitbody

Dilution Ratio1:500

Secondary Antibody

Secondary Descriptiongoat anti-rabbit
Immunohistochemistry-Paraffin Nox4 NB110-58849
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reviewed by:
Anonymous
IHC-P Mouse 01/30/2013
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Summary

ApplicationImmunohistochemistry-Paraffin
Sample Testedmouse brain
SpeciesMouse
CommentsThe Nox4 antibody showed positive staining in some neurons in the mouse cortex in our knockout mice. Very little expression was detected in control mice. It seemed that NOX4 was relatively specific, but though it was used at 1:50, it seemed a little weak.

Blocking

Blocking DetailsBlocked in PBS with 0.5% Triton-X and 10% goat serum

Primary Anitbody

Dilution Ratio1:50 Dilution, incubated overnight at 4 degrees. PBS with 0.5% Tween-80 and 10% goat serum.

Secondary Antibody

Secondary DescriptionAlexa Fluor 555 Goat anti-rabbit IgG
Secondary Manufacturer Cat#Life Technologies A-21428
Secondary Concentration1:250

Details

Detection NotesFluorescent microscope
Fixation DetailsFixed in paraformaldehyde, embedded in parafiin, antigen retrieval with sodium citrate buffer
Wash DescriptionWashed in PBS

Comments

CommentsThe Nox4 antibody showed positive staining in some neurons in the mouse cortex in our knockout mice. Very little expression was detected in control mice. It seemed that NOX4 was relatively specific, but though it was used at 1:50, it seemed a little weak.
Western Blot Nox4 NB110-58849
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reviewed by:
Vivian Lin
WB Human 12/10/2012
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Summary

ApplicationWestern Blot
Sample TestedHUVEC whole cell lysate
SpeciesHuman
LotE
CommentsLanes 1 and 2: serum-starved HUVEC lysate denatured at 95 oC for 5 minLanes 3: serum-starved HUVEC lysate denatured at RT for 10 min

Blocking

Blocking Details4% BSA/TBST for 2 hr at RT

Primary Anitbody

Dilution Ratio1:500 in 4% BSA/TBST, overnight 2-8 oC

Secondary Antibody

Secondary Descriptiongoat anti-rabbit IgG, HRP
Secondary Manufacturer Cat#sc-2004
Secondary Concentration1:5000 in 4% BSA/TBST, 1 hr at RT

Details

Detection NotesWestern Lighting Plus ECL reagent, 25 s exposure

Comments

CommentsLanes 1 and 2: serum-starved HUVEC lysate denatured at 95 oC for 5 minLanes 3: serum-starved HUVEC lysate denatured at RT for 10 min
Western Blot Nox4 NB110-58849
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reviewed by:
Anonymous
WB Human 11/15/2012
View

Summary

ApplicationWestern Blot
Sample TestedIMR90 fibroblast, +/- TGF beta, whole cell lysate
SpeciesHuman
LotE4, E6, E7
CommentsAll lot numbers detect a band just above the 64 kD marker (red arrow) corresponding to Nox4 isoform 1 (~67kD) as evidenced by strong induction upon TGF β1 treatment. Some lots also detect a band at ~55 kD. Background and nonspecific bands can be reduced by optimizing for each lot number. Overall excellent antibody, our lab’s “gold standard” for Nox4.

Blocking

Blocking DetailsTBST + 5% milk OR SuperBlock T20 (Thermo Fisher)

Primary Anitbody

Dilution Ratio1:2000 or 1:5000 in TBST + 5% milk, TBST + 5% BSA, or SuperBlock, overnight at 4C

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG (H+L)-HRP
Secondary Manufacturer Cat#Pierce #31462
Secondary Concentration1:10,000 in milk or SuperBlock

Details

Detection NotesMillipore Luminata Chemiluminescent HRP substrates as listed, expose to film

Comments

CommentsAll lot numbers detect a band just above the 64 kD marker (red arrow) corresponding to Nox4 isoform 1 (~67kD) as evidenced by strong induction upon TGF β1 treatment. Some lots also detect a band at ~55 kD. Background and nonspecific bands can be reduced by optimizing for each lot number. Overall excellent antibody, our lab’s “gold standard” for Nox4.

Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for Nox4 Antibody (NB110-58849). (Showing 1 - 3 of 3 FAQs).

  1. Do you have an alternative product for NOX4 Antibody (NB110-58851). I will be using it for porcine.
    • NB110-58849 is validated in the same species (apart from porcine) and applications as NB110-58851 and should be a suitable alternative for you. NB110-58851 was raised to a synthetic peptide made to a C-terminal region (within residues 500-578) of the human NOX4 protein, whilst NB110-58849 was raised to a synthetic peptide made to an internal region (between residues 100-200) of the human NOX4 protein. Both antibodies are rabbit polyclonals. As NB110-58849 has not yet been tested in porcine samples, you may be interested in our Innovator's Reward programme. This allows you to try our primary antibodies in an untested species or application, without the financial risk of failure. To participate you simply need to go to the antibody’s webpage and complete an online review with an image, detailing the positive or negative results of your study. In return you will receive a discount voucher for 100% of the purchase price of the reviewed product. More details of this programme can be found here: Innovator's Reward Program. I ran an alignment for you between the human NOX4 sequence and an unreviewed porcine NOX4 sequence from Uniprot (accession number F1STQ7.). The homology is high between the two proteins. Please do, however, be aware that this porcine sequence is unreviewed.
  2. Is NB110-58849PEP the exact peptide sequence you raised your polyclonal antibody against? And, do you have any Westerns or IHC images with your NOX4 antibody combined with this as a blocking peptide and showing very little background?
    • We have used this blocking peptide in our testing in the past; however, I am unable to find any available images on our system. NOX4 does tend to be difficult to work with in WB, as there are 9 different isoforms of the protein. However, we will guarantee that the blocking peptide and the antibody will perform together.
  3. What is the molecular weight of NOX4 (for western blot)?
    • The MW of this protein is Length:578 Mass (Da):66,932 on Uniprot. There are Application Notes for this antibody: In Western blot this antibody recognizes a band at ~67 kDa or larger representing isoform 1 of NOX4, and what appears to be a non-specific band ~48 kDa. The observed band size may vary depending on sample type and glycosylation.

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB110-58849AF405
Alexa Fluor 488 Labeled NB110-58849AF488
Alexa Fluor 647 Labeled NB110-58849AF647
Alexa Fluor 700 Labeled NB110-58849AF700
Biotin Labeled NB110-58849B
DyLight 405 Labeled NB110-58849V
DyLight 488 Labeled NB110-58849G
DyLight 550 Labeled NB110-58849R
DyLight 650 Labeled NB110-58849C
FITC Labeled NB110-58849F
HRP Labeled NB110-58849H

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Blogs on Nox4.

NOX4 Antibodies: Don't NOX them until you've tried them
NOX4 is an NADPH oxidase that generates superoxide within the cell. It is primarily found in vascular cells, fibroblasts, and osteoclasts, with abundant expression in the kidney. Unlike its family members NOX1 and NOX2, NOX4 is constitutively active,...  Read full blog post.

NOX4 Antibodies in Diabetic Nephropathy Research
Diabetic nephropathy (DN) is one of the leading complications resulting from chronic diabetes. It manifests as progressive renal failure caused by mesangial cell hyperplasia and fibrosis, and is one of the leading causes of terminal kidney disease (1)...  Read full blog post.

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Recent Reviews

4.4
5
3
4
4
3
0
2
0
1
0

Anonymous
03/01/2018
Application: WB
Species: Rat

Anonymous
07/25/2016
Application: IF
Species: Other

Anonymous
07/22/2016
Application: WB
Species: Other

Bioinformatics

Gene Symbol NOX4