P47phox

Manufactured by Merck Group

Sourced in United States, United Kingdom

P47phox is a laboratory equipment product. It is a protein that plays a core role in the activation of the NADPH oxidase enzyme complex, which is responsible for the generation of superoxide radicals in phagocytic cells. The product specifications and intended use are not available for this description.

Automatically generated - may contain errors

Lab products found in correlation

Gp91phox, by BD (4 mentions) Interleukin 6 (il 6), by Merck Group (2 mentions) P22phox, by Santa Cruz Biotechnology (2 mentions) Phospho src, by Cell Signaling Technology (2 mentions) Phospho erk1 2, by Cell Signaling Technology (2 mentions) Gapdh, by Abcam (2 mentions) P67phox, by Merck Group (2 mentions) β actin, by Santa Cruz Biotechnology (2 mentions) Sodium diethyldithiocarbamate, by Merck Group (1 mentions) Dihydroethidium, by Thermo Fisher Scientific (1 mentions) Glutathione ethyl ester, by Merck Group (1 mentions) Dithiothreitol (dtt), by Promega (1 mentions) Osmotic minipumps, by Merck Group (1 mentions) Diethylenetriaminepentaacetic acid, by Merck Group (1 mentions) Streptavidin sepharose, by GE Healthcare (1 mentions) Feso4 7h2o, by Merck Group (1 mentions) Protein a g plus agarose, by Santa Cruz Biotechnology (1 mentions) α tubulin, by Santa Cruz Biotechnology (1 mentions) Recombinant ck2, by New England Biolabs (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions)

Spelling variants of this product

Anti-p47phox (9 citations) Rabbit anti-p47phox (3 citations) P47phox antibody (2 citations) Phospho-p47phox antibody (2 citations) Anti-p47phox antibody (2 citations)

20 protocols using p47phox

Biochemical Assay Reagents and Antibodies

Check if the same lab product or an alternative is used in the 5 most similar protocols

Sodium diethyldithiocarbamate, FeSO₄ 7H₂O, diethylenetriaminepentaacetic acid, and glutathione ethyl ester were purchased from Sigma‐Aldrich. N‐Hydroxysulfosuccinimidobiotin was obtained from Merck, dithiothreitol from Promega, streptavidin–Sepharose from GE Healthcare Bio‐Sciences, and Protein A/G‐Plus agarose from Santa Cruz Biotechnology. Osmotic mini‐pumps were purchased from Alzet and CL from Sigma‐Aldrich. Dihydroethidium and N‐(Biotinoyl)‐N′‐(iodoacetyl) ethylenediamine were obtained from Invitrogen. Monoclonal antibodies were purchased from the following vendors: β₁ subunit of Na⁺‐K⁺ ATPase from Upstate Biotechnology; anti‐glutathione from Virogen, eNOS antibody from Sigma‐Aldrich, and p47^phox, phosphorylated eNOS116, phosphorylated eNOS1177, neuronal NOS (nNOS), and α tubulin from Santa Cruz Biotechnology. All chemicals used in Krebs solutions were analytical grade and were obtained from BDH.

Karimi Galougahi K., Liu C., Garcia A., Gentile C., Fry N.A., Hamilton E.J., Hawkins C.L, & Figtree G.A. (2016). β3 Adrenergic Stimulation Restores Nitric Oxide/Redox Balance and Enhances Endothelial Function in Hyperglycemia. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 5(2), e002824.

+ Open protocol

+ Expand

CK2 Phosphorylation Assay on Immobilized Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Ten or five picomoles of recombinant NOXO1β, p47^PHOX, NOXA1, p67^PHOX, and BSA (Sigma-Aldrich, Lyon, France) were applied directly on nitrocellulose membrane in a single spot and let to air dry. The membranes were blocked for 1 hour at room temperature in TBS/T (20 mmol/L Tris-HCl [pH 7.6], 137 mmol/L NaCl, 0.1% Tween 20) containing 5% nonfat dry milk and then incubated with 1 μg/mL recombinant CK2 (New England Biolabs) diluted in TBS/T containing 1% fat dry milk overnight at 4°C. After several washes with TBS/T, bound CK2 was detected with a mouse monoclonal anti-CK2 α/α′ antibody at 1/1000 dilution for 1 hour at room temperature. Antibody binding was detected using horseradish peroxidase–conjugated anti-mouse IgG. Blots were visualized using ECL Western blotting reagents.

Liu D., Marie J.C., Pelletier A.L., Song Z., Ben-Khemis M., Boudiaf K., Pintard C., Leger T., Terrier S., Chevreux G., El-Benna J, & Dang P.M. (2022). Protein Kinase CK2 Acts as a Molecular Brake to Control NADPH Oxidase 1 Activation and Colon Inflammation. Cellular and Molecular Gastroenterology and Hepatology, 13(4), 1073-1093.

+ Open protocol

+ Expand

Immunostaining of Brain Slices for Oxidative Stress Markers

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Frozen brain slices and microglia-bearing coverslips mentioned above were immunostained as previously described [80 (link)]. Briefly, after incubation with primary antibodies of p47^phox (1:200, Sigma-Aldrich, St. Louis, MO, USA), PI3Kγ (1:500, Novus, Littleton, CO, USA), Iba1 (1:500, Millipore, Billerica, MA, USA) and NeuN (1:500, Millipore, Billerica, MA, USA), antibody binding was visualized with Alexa Fluor 488-labeled goat anti-mouse IgG and Cy3-labeled goat anti-rabbit IgG (1:200, both from Beyotime Biotechnology, Shanghai, China). Cell nuclei were counterstained with 4’,6-diamidino-2-phenylindole (DAPI). Images of the penumbral regions were obtained using a confocal microscope. The Pearson’s correlation coefficient and the numbers of antibody-positive cells were analyzed by a blinded investigator using Image J.

Wang L., Wang X., Li T., Zhang Y, & Ji H. (2018). 8e Protects against Acute Cerebral Ischemia by Inhibition of PI3Kγ-Mediated Superoxide Generation in Microglia. Molecules, 23(11), 2828.

+ Open protocol

+ Expand

Phosphorylation of p47phox Protein

Check if the same lab product or an alternative is used in the 5 most similar protocols

The immunoprecipitation assays were performed as previously described (Lang et al., 2000 ). Liver proteins were precipitated with polyclonal antibodies against p47^phox (Santa Cruz Biotechnology, Santa Cruz, CA). Immunocomplexes were recognized by western blot using specific antibodies against p47^phox and anti-phosphoserine (Sigma-Aldrich).

García-Ruiz I., Solís-Muñoz P., Fernández-Moreira D., Grau M., Colina F., Muñoz-Yagüe T, & Solís-Herruzo J.A. (2014). High-fat diet decreases activity of the oxidative phosphorylation complexes and causes nonalcoholic steatohepatitis in mice. Disease Models & Mechanisms, 7(11), 1287-1296.

+ Open protocol

+ Expand

siRNA Transfection for Gene Silencing

Check if the same lab product or an alternative is used in the 5 most similar protocols

We purchased the human siRNAs of scrambled, Nox2, MMP-9, p47^phox, p65, IL-6, Nox4, AT1R, and AT2R from Sigma (St. Louis, MO). Transient transfections with various siRNAs (100 nM) were performed using Lipofectamine transfection reagent in accordance with the manufacturer's protocols. The siRNA used in this study were labeled with EGFP. The transfection efficiency was evaluated by transfection with EGFP, and was determined to be approximately 60%.

Tsai M.H., Lee C.W., Hsu L.F., Li S.Y., Chiang Y.C., Lee M.H., Chen C.H., Liang H.F., How J.M., Chang P.J., Wu C.M, & Lee I.T. (2017). CO-releasing molecules CORM2 attenuates angiotensin II-induced human aortic smooth muscle cell migration through inhibition of ROS/IL-6 generation and matrix metalloproteinases-9 expression. Redox Biology, 12, 377-388.

+ Open protocol

+ Expand

Protein Expression Analysis in FDB Muscles

Check if the same lab product or an alternative is used in the 5 most similar protocols

FDB muscles were homogenized using an electric homogenizer (Fluko, Shanghai, China) in a lysis buffer containing in mM: 20Tris-HCl (pH 7.5), 1% Triton X-100, 2 EDTA, 20 NaF, 1 Na₂P₂O₇, 10% glycerol, 150 NaCl, 10 Na₃VO₄, 1 PMSF and protease inhibitors (Complete™, Roche Applied Science). Proteins were separated using SDS-PAGE and transferred to PVDF membranes. The following primary antibodies and their dilutions were used as follows: Total NF-κB total p65 subunit (1:1000; Cell Signaling); p-Ser536-p65 (1:1000; Cell Signaling); p47^phox (1:5000; Sigma); p-p47^phox (pSer359) (1:5000; Sigma); gp91^phox (1:2000; BD Transduction); p-p38 (thr180/tyr182) (1:2000; Santa Cruz); p38 (1:2000; Santa Cruz); Phospho-ERK1/2(Thr202/Tyr204)(1:2000; Cell Signaling); ERK 1/2 (1:2000; Santa Cruz); α-Tubulin (1:5000; Cell Signaling); anti-mouse IgG-HRP (1:20,000; Santa Cruz); anti-rabbit IgG-HRP (1:30,000; Thermo Scientific Pierce). The protein bands in the blots were visualized using a WESTAR Supernova detection kit (Cyanagen, Bologna, Italy) and ChemiDoc™ MP System (Bio-Rad, USA). The intensity of the bands was determined with ImageJ densitometry analysis.

Henríquez-Olguín C., Díaz-Vegas A., Utreras-Mendoza Y., Campos C., Arias-Calderón M., Llanos P., Contreras-Ferrat A., Espinosa A., Altamirano F., Jaimovich E, & Valladares D.M. (2016). NOX2 Inhibition Impairs Early Muscle Gene Expression Induced by a Single Exercise Bout. Frontiers in Physiology, 7, 282.

+ Open protocol

+ Expand

Western Blot Analysis of Protein Signaling

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

PC12 cells and primary neurons, or PC12 cells infected with lentiviral shRNA to NOX2 or GFP, or PC12 cells infected with Ad-PP5, Ad-dn-c-Jun or Ad-GFP, respectively, after treatments, were lysed, followed by Western blotting, as described previously (Chen et al., 2010 (link); Chen et al., 2014a (link)). The blots for detected protein were semi-quantified using NIH Image J software (National Institutes of Health, Bethesda, MD, USA). The following antibodies were used: phosphorylated JNK (p-JNK) (Thr183/Tyr185), JNK, p-c-Jun (Ser63), c-Jun, p22^phox (Santa Cruz Biotechnology, Santa Cruz, CA, USA), caspase-3, and PARP (Cell Signaling Technology, Beverly, MA, USA), PP5 (BD Biosciences, San Jose, CA), NOX2, p67^phox (Epitomics, Burlingame, CA, USA), Rac1 (Cytoskeleton, Denver, CO, USA), p40^phox, p47^phox, FLAG, HA, and β-tubulin (Sigma), goat anti-rabbit IgG-horseradish peroxidase (HRP), goat anti-mouse IgG-HRP, and rabbit anti-goat IgG-HRP (Pierce, Rockford, IL, USA).

Xu C., Wang X., Gu C., Zhang H., Zhang R., Dong X., Liu C., Hu X., Ji X., Huang S, & Chen L. (2017). Celastrol ameliorates Cd-induced neuronal apoptosis by targeting NOX2-derived ROS-dependent PP5-JNK signaling pathway. Journal of neurochemistry, 141(1), 48-62.

+ Open protocol

+ Expand

Transient Knockdown of Signaling Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Scrambled, ICAM-1, IL-6, p47^phox, JNK2, p42, p38, p65, p50, TLR2, and TLR4 human siRNAs were purchased from Sigma (St. Louis, MO). We transiently transfected siRNA (100 nM) using a Lipofectamine^® 2000 Reagent according to the manufacturer's instructions.

Lee C.W., Wu C.H., Chiang Y.C., Chen Y.L., Chang K.T., Chuang C.C, & Lee I.T. (2018). Carbon monoxide releasing molecule-2 attenuates Pseudomonas aeruginosa-induced ROS-dependent ICAM-1 expression in human pulmonary alveolar epithelial cells. Redox Biology, 18, 93-103.

+ Open protocol

+ Expand

Targeted knockdown of signaling proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human siRNAs of PKCα, p47^phox, p38, JNK1, and scrambled were from Sigma (St. Louis, MO). Transient transfection of siRNAs was performed using Metafectene transfection reagent from Biontex Lab (GmbH, Planegg/Martinsried, Germany) according to the manufacturer's instructions. The transfection mixture was diluted into 500 μL of DMEM/F-12 medium with 10% FBS and antibiotics and added directly to the cells. After 16 h transfection, the medium was replaced with serum-free DMEM/F-12 for 24 h. Cell lysates prepared from RASFs challenged with TNF-α were analyzed by Western blot.

Chi P.L., Liu C.J., Lee I.T., Chen Y.W., Hsiao L.D, & Yang C.M. (2014). HO-1 Induction by CO-RM2 Attenuates TNF-α-Induced Cytosolic Phospholipase A2 Expression via Inhibition of PKCα-Dependent NADPH Oxidase/ROS and NF-κB. Mediators of Inflammation, 2014, 279171.

+ Open protocol

+ Expand

siRNA-mediated Knockdown of Key Signaling Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

All human ASK1, p47^phox, TRAF2, NIK, IKKα, IKKβ, and p65 siRNA together with scramble siRNA were purchased from Sigma (St. Louis, MO, USA). Lipofectamine^TM RNAiMAX reagents were used to prepare siRNA liposome complexes (100 nM of siRNAs) according to the manufacturer’s instructions, as previously described Lee et al. (2008) (link).

Lin C.C., Lin W.N., Cho R.L., Wang C.Y., Hsiao L.D, & Yang C.M. (2016). TNF-α-Induced cPLA2 Expression via NADPH Oxidase/Reactive Oxygen Species-Dependent NF-κB Cascade on Human Pulmonary Alveolar Epithelial Cells. Frontiers in Pharmacology, 7, 447.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!