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P40phox

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P40phox is a protein that plays a critical role in the activation of the NADPH oxidase complex, which is responsible for the production of reactive oxygen species (ROS) in phagocytic cells. It serves as an essential regulatory subunit of the NADPH oxidase complex, contributing to its assembly and activation.

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Lab products found in correlation

P22phox, by Santa Cruz Biotechnology (4 mentions) P47phox, by Santa Cruz Biotechnology (3 mentions) P67phox, by Santa Cruz Biotechnology (3 mentions) Dihydroethidium dhe, by Thermo Fisher Scientific (3 mentions) Nitrotyrosine, by Santa Cruz Biotechnology (1 mentions) Gp91phox, by BD (1 mentions) Phospho enos ser1177, by Cell Signaling Technology (1 mentions) Lysis buffer, by Cell Signaling Technology (1 mentions) Protease inhibitor cocktail, by Cell Signaling Technology (1 mentions) P p40phox, by Cell Signaling Technology (1 mentions) Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions) Imagequant las 4000, by GE Healthcare (1 mentions) Polyacrylamide gel, by Bio-Rad (1 mentions) Westernbright quantum, by Advansta (1 mentions) Gp91phox, by Santa Cruz Biotechnology (1 mentions) Apocynin, by Merck Group (1 mentions) N nitro l arginine methyl ester l name, by Merck Group (1 mentions) P enos, by Abcam (1 mentions) Ecl developer, by Merck Group (1 mentions) β actin, by Cell Signaling Technology (1 mentions)

5 protocols using p40phox

1

Protein Extraction and Antibody Detection in Myocardial Tissues

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The preparation of protein samples from myocardial tissues was performed as previously described (Becker et al. 2005 (link)). Antibodies to eNOS (BD Transduction Laboratories, 1:250 dilution), phospho-eNOS (Ser 1177) (Cell Signal, 1:500 dilution), SOD-1(Calbiochem, 1:5000 dilution), SOD-2 (BD Transduction Laboratories, 1:10,000 dilution), SOD-3 (Santa Cruz Biotechnology, 1:5000 dilution), or one of the following subunits of NAD(P)H oxidase: p67phox (Upstate, 1:1000 dilution), p22phox (Santa Cruz Biotechnology, 1:2000 dilution), gp91phox(BD Transduction Laboratories, 1:1000 dilution), p47phox (Santa Cruz Biotechnology, 1:1000 dilution), phos-p47phox (Upstate, 1:1000 dilution), p40phox (Santa Cruz Biotechnology, 1:800 dilution), Rac-1 (Santa Cruz Biotechnology, 1:5000 dilution), and nitrotyrosine (Santa Cruz Biotechnology, 1:1000 dilution) were used.
Song S., Kertowidjojo E., Ojaimi C., Martin-Fernandez B., Kandhi S., Wolin M, & Hintze T.H. (2015). Long-term methionine-diet induced mild hyperhomocysteinemia associated cardiac metabolic dysfunction in multiparous rats. Physiological Reports, 3(5), e12292.
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2

Western Blotting Analysis of Cell Signaling

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Cells were lysed in ice-cold lysis buffer (Cell Signaling Technology), 1mM phenylmethylsulfonyl fluoride (PMSF), and protease inhibitor cocktail (Cell Signaling Technology). Cell lysate were cleared by rapid centrifugation for 5 min at 4° C. Equal amounts of proteins were separated by SDS-PAGE on a 10% or 12% polyacrylamide gel (Bio-Rad), transferred to polyvinylidene difluoride (PVDF) membrane, blocked with 5% BSA (Fisher, Pittsburgh PA) in TBS-T (25 mM Tris, 0.15M NaCl, 0.05% Tween-20, pH 7.5). Primary antibodies were detected using horseradish peroxidase (HRP)-conjugated secondary antibodies and a chemiluminescent detection reagent (Western Bright Quantum [Advansta]). Antibodies to pAkt, Akt, p-p40phox, Ras and GRB2 were purchased from Cell Signaling Technology and p40phox from Santa Cruz Biotechnology. Blots were imaged using a GE ImageQuant LAS 4000 digital imaging system. Scanned images were processed for brightness and contrast using only the levels function of Adobe Photoshop applied to the entire image before cropping.
Vareechon C., Zmina S.E., Karmakar M., Pearlman E, & Rietsch A. (2017). Pseudomonas aeruginosa effector ExoS inhibits ROS production in human neutrophils. Cell host & microbe, 21(5), 611-618.e5.
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3

Molecular Mechanism of Herb-Mediated Vascular Protection

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The herbs were provided by Shanghai Huayu Chinese Herbs Co. Ltd. (Shanghai, China). Dihydroethidium (DHE) was purchased from Molecular Probes Inc. (Eugene, USA). Antibodies of p47phox, p67phox, p40phox, p22phox, and gp91phox were from Santa Cruz Biotechnology (Santa Cruz, CA, USA); Ras-1 and β-actin were obtained from Cell Signaling Technology (Danvers, MA, USA); eNOS and p-eNOS were provided by Abcam (Cambridge, MA, USA). Goat anti-rabbit secondary antibodies were bought from Wuhan Boster Biotech Co. Ltd. (Wuhan, China). ECL developer was from Millipore (Billerica, MA, USA). BCA protein quantity kits and PVDF membranes were provided by Pierce (Rockford, IL, USA). Phenylephrine (PE), acetylcholine (ACh), Hcy, sodium nitroprusside (SNP), tempol, apocynin, and N-nitro-L-arginine methyl ester (L-NAME) were purchased from Sigma Chemical Co. Ltd. (St. Louis, MO, USA). All reagents available were of high purity.
Chu S., Mao X.D., Wang L, & Peng W. (2016). Effects of Huang Qi Decoction on Endothelial Dysfunction Induced by Homocysteine. Evidence-based Complementary and Alternative Medicine : eCAM, 2016, 7272694.
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4

Antioxidant Compound Synthesis and Validation

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LMH001 (2,3,-dihydroxyphenyl) methyl 4-hydroxy-3-(hydroxyrnethyl) benzoate, MW = 290.079) (molecular grade) was synthesized at Tocris Biosciences (Bristol, UK) [15 ]. AngII was purchased from Sigma-Aldrich. Polyclonal antibodies against Nox1, Nox2, Nox4, p22phox, p40phox, p47phox, p67phox, rac1 and CD31 were from Santa Cruz Biotechnology. Antibodies to mouse CD45 and CD68 were from Abcam (UK). Antibodies to phospho-ERK1/2 (ERK1Thr202/Tyr204 and ERK2Thr185/Tyr187), phospho-p38MAPKThr180/Tyr182, and phospho-JNKThr183/Tyr185, Thr221/Tyr223 were from Sigma-Aldrich (UK). DHE (dihydroethidium) was purchased from Invitrogen (UK). The scrambled control peptide and Nox2tat ([H]-RKKRRQRRRCSTRVRRQL-[NH2]) were provided by PeptideSynthetics (PPR Ltd. Fareham, UK). All other reagents and chemicals were from Sigma-Aldrich (UK) unless stated.
Fan L.M., Liu F., Du J., Geng L, & Li J.M. (2022). Inhibition of endothelial Nox2 activation by LMH001 protects mice from angiotensin II-induced vascular oxidative stress, hypertension and aortic aneurysm. Redox Biology, 51, 102269.
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5

Antibody Validation for Oxidative Stress

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Polyclonal antibodies against Nox1, Nox2, Nox4, p22phox, p40phox, p47phox, p67phox, rac1, insulin receptors (IRα and IRβ) were from Santa Cruz Biotechnology. Antibodies to phospho-ERK1/2, phospho-p38MAPK, phospho-JNK and phospho-Aktser473 were from Cell Signaling Technology. DHE (dihydroethidium) and 5-(and 6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate (DCF) were from Invitrogen (UK). All other reagents and chemicals were from Sigma unless stated otherwise.
Fan L.M., Cahill-Smith S., Geng L., Du J., Brooks G, & Li J.M. (2017). Aging-associated metabolic disorder induces Nox2 activation and oxidative damage of endothelial function. Free Radical Biology & Medicine, 108, 940-951.
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