Anti mnsod

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-MnSOD is a laboratory product that detects the manganese superoxide dismutase (MnSOD) protein. MnSOD is an enzyme responsible for scavenging superoxide radicals in the mitochondria. This antibody can be used to measure the expression levels of MnSOD in various biological samples.

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Close spelling variants of this product

MnSOD (29 citations) Goat anti-MnSOD-1:100 (2 citations)

14 protocols using anti mnsod

Leelamine Modulates CXCR7/CXCR4 Signaling

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Leelamine (LEE, Figure 1A) was purchased from Cayman Chemical (Ann Arbor, MI, USA). LEE stock solution (10 mM) was prepared in EtOH, storage at −20 °C and finally diluted in cell culture medium for use. Anti-CXCR7 and anti-CXCR4 antibodies were purchased from abcam (Cambridge, UK). Anti-MnSOD, anti-fibronectin, anti-vimentin, anti-MMP-9, anti-MMP-2, anti-N-cadherin, anti-E-cadherin, anti-twist, anti-snail, anti-occludin, and anti-b-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-phospho-STAT3(Tyr705), anti-STAT3, anti-phospho-JAK1(Tyr1022/1023), anti-JAK1, anti-phospho-JAK2(Tyr1007/1008), and anti-JAK2 were purchased from Cell Signaling Technology (Beverly, MA, USA). Alexa Fluor^® 488 donkey anti-rabbit IgG (H+L) antibody and Alexa Fluor^® 594 donkey anti-mouse IgG (H+L) antibody was obtained from Life Technologies (Grand Island, NY, USA). The GSH/GSSG-Glo™ Assay kit was purchased from Promega (Madison, WI, USA).

Jung Y.Y., Um J.Y., Sethi G, & Ahn K.S. (2022). Potential Application of Leelamine as a Novel Regulator of Chemokine-Induced Epithelial-to-Mesenchymal Transition in Breast Cancer Cells. International Journal of Molecular Sciences, 23(17), 9848.

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Green Tea Polyphenols Therapeutic Study

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GTPs (91.21% catechins and 71.72% EGCG) were purchased from Corona Science & Technology Development Co. Ltd. (Fu Zhou, China). Rabbit polyclonal antibodies (anti-MnSOD, anti-HMGCS2, anti-Nampt, and anti-catalase) were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, USA). Anti-FOXO3a and anti-SIRT3 antibodies were from Cell Signaling Technology Inc. (Danvers, USA). Anti-SOD2/MnSOD (acetyl K122) and anti-4-HNE antibodies were from Abcam Inc. (Cambridge, UK). Total cholesterol (TC), triglycerides (TG), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), and blood glucose were detected using kits from BioSino Bio-Technology & Science Inc. (Beijing, China). ELISA kits for the quantitative measurement of serum cystatin C and insulin were from Biovendor Inc. (Heidelberg, Germany) and Mercodia AB (Uppsala, Sweden). Creatinine and N-acetyl-β-D-glucosaminidase (NAG) assay kits were purchased from the Nanjing Jiancheng Bioengineering Institute (Jiangsu, China). DMEM/high glucose was purchased from Gibco Inc. (New York, USA). Lipofectamine 3000 transfection reagent was from Invitrogen Inc. (California, USA).

Yi W., Xie X., Du M., Bu Y., Wu N., Yang H., Tian C., Xu F., Xiang S., Zhang P., Chen Z., Zuo X, & Ying C. (2017). Green Tea Polyphenols Ameliorate the Early Renal Damage Induced by a High-Fat Diet via Ketogenesis/SIRT3 Pathway. Oxidative Medicine and Cellular Longevity, 2017, 9032792.

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Corilagin Attenuates EMT and Wnt Signaling in Cancer

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Corilagin (CLG) was procured from Sigma-Aldrich (St. Louis, MO). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and bovine serum albumin (BSA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Alexa Fluor^® 488 donkey anti-mouse IgG (H+L) antibody and Fluor^® 594 donkey anti-rabbit IgG (H+L) antibody was obtained from Life Technologies (Grand Island, NY, USA). Anti-MnSOD(sc-137254), anti-Fibronectin(sc-6952), anti-Vimentin(sc-6260), anti-E-cadherin(sc-8426), anti-N-cadherin(sc-271386), anti-Occludin(sc-5562), anti-Twist(sc-15393), anti-MMP-2(sc-53630), anti-MMP-9(sc-393859), anti-Wnt3a(sc-136163), anti-FZD-1(sc-398082), and anti-β-actin(sc-47778) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-Snail(3879S), anti-Axin-1 anti-Azin-1(3323S), anti-β-catenin(9562S), anti-p-GSK3β(9322S), and anti-GSK3β(9315S) antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA).

Hwang S.T., Yang M.H., Kumar A.P., Sethi G, & Ahn K.S. (2020). Corilagin Represses Epithelial to Mesenchymal Transition Process Through Modulating Wnt/β-Catenin Signaling Cascade. Biomolecules, 10(10), 1406.

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Antioxidant Enzyme Expression Profiling

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APN, SCO, and other chemical reagents were obtained from Sigma-Aldrich (MO, USA). Anti- MnSOD, anti-CuZnSOD, anti-GAPDH, and anti-NF-E2-related factor 2 (Nrf2) antibodies were provided by Santa Cruz Biotechnology (TX, USA). Anti-HO-1 and anti-pphospho-Nrf2 (p-Nrf2) antibodies were supplied from Stressgen (MI, USA) and Epitomics (CA, USA), respectively.

Lee G.Y., Lee C., Park G.H, & Jang J.H. (2017). Amelioration of Scopolamine-Induced Learning and Memory Impairment by α-Pinene in C57BL/6 Mice. Evidence-based Complementary and Alternative Medicine : eCAM, 2017, 4926815.

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Immunohistochemical Analysis of Oxidative Stress and Inflammatory Markers

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Pancreas and lung sections were incubated with the following primary antibodies: Anti-NRF2 (sc-365949, 1:200, Santa Cruz Biotechnology, CA, USA); anti-HO-1 (sc-136960, 1:200, Santa Cruz Biotechnology, CA, USA); anti-Mn-SOD (sc-137254, 1:200, Santa Cruz Biotechnology, CA, USA); anti-NLRP3 (sc-134306, 1:200, Santa Cruz Biotechnology, CA, USA); anti-Caspase-1 (sc-56036, 1:200, Santa Cruz Biotechnology, CA, USA); and anti-ASC (sc-514414, 1:200, Santa Cruz Biotechnology, CA, USA), as previously described [48 (link)]. Sections were then incubated with the following secondary antibodies: Peroxidase-conjugated bovine anti-mouse immunoglobulin G (IgG) or peroxidase-conjugated goat anti-rabbit IgG (1:2000, Jackson Immuno Research, West Grove, PA, USA). Specific marking was revealed with a biotin-conjugated goat anti-rabbit IgG or biotin-conjugated goat anti-mouse IgG and avidin-biotin peroxidase complex (Vector Laboratories, Burlingame, CA, USA). Graphic presentation of densitometric analyses was performed Image J software (v1.52a) as previously described [49 (link)]. All immunohistochemical analyses were conducted by an observer without knowledge of the treatments.

Cordaro M., Fusco R., D’Amico R., Siracusa R., Peritore A.F., Gugliandolo E., Genovese T., Crupi R., Mandalari G., Cuzzocrea S., Di Paola R, & Impellizzeri D. (2020). Cashew (Anacardium occidentale L.) Nuts Modulate the Nrf2 and NLRP3 Pathways in Pancreas and Lung after Induction of Acute Pancreatitis by Cerulein. Antioxidants, 9(10), 992.

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Evaluating Cellular Antioxidant Markers in H9c2 Cells

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The H9c2 cells were cultured at 4.5×10⁵ cells/well in 6-well plates and allowed to grow for 24 h. They were then treated as described above to determine the CAT, MnSOD, p-IKB and nitrotyrosine levels. The H9c2 cells were collected using a scraper, treated with fixing buffer (4% formaldehyde, 0.1% NaN₃, 2% FBS, in PBS) for 20 min, and then permeabilized using a fixation and permeabilization buffer (fixing buffer containing 0.1% Triton X) for 30 min. Subsequently, these cells were incubated with the anti-CAT (diluted 1:250; cat. no. sc-271803; Santa Cruz Biotechnology, Inc.), anti-MnSOD (diluted 1:250; cat. no. sc-137254; Santa Cruz Biotechnology, Inc.), anti-p-IKB and anti-nitrotyrosine antibodies (both diluted 1:250; cat. no. sc-137254 and sc-32757 respectively; Santa Cruz Biotechnologies) for 1 h at 4°C, as required, followed by the respective anti-rabbit or anti-mouse FITC antibodies as the secondary antibodies (diluted both 1:5,000; cat. nos. A120-208F and A90-146F, respectively; Bethyl Laboratories), for 1 h at 4°C. The cells were collected and analyzed by FACS (FACSscan; BD Biosciences) and the data obtained were processed using the CellQuest software, version 5.2.1. These data are shown as percentages of positive cells.

Pecoraro M., Pala B., Di Marcantonio M.C., Muraro R., Marzocco S., Pinto A., Mincione G, & Popolo A. (2020). Doxorubicin-induced oxidative and nitrosative stress: Mitochondrial connexin 43 is at the crossroads. International Journal of Molecular Medicine, 46(3), 1197-1209.

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Antibody Selection for Molecular Analysis

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Monoclonal antibodies specific for Beclin-1 and p62 were purchased from Sigma-Aldrich (Saint Louis, USA). Anti-GAPDH monoclonal antibody was acquired from Calbiochem-Merck Chemicals Ltd. (Nottingham, UK). Similarly, anti-NLRP3 antibody was purchased from Adipogen (San Diego, USA); while anti-active caspase-3, anti-SIRT-1 and anti-Parkin were obtained from Cell Signalling Technology (Beverly, MA, USA). Finally, anti-IL-1β (p17), anti-OGG-1, anti-Bcl-2, anti-Bax, anti-MnSOD, anti-catalase, anti-ATG12 and anti-MAP-LC3 antibodies from (Santa Cruz Biotechnology). A cocktail of protease inhibitors (Complete™ Protease Inhibitor Cocktail) was purchased from Boehringer Mannheim (Indianapolis, IN). The Immun Star HRP substrate kit was obtained from Bio-Rad Laboratories Inc. (Hercules, CA).

Pavillard L.E., Cañadas-Lozano D., Alcocer-Gómez E., Marín-Aguilar F., Pereira S., Robertson A.A., Muntané J., Ryffel B., Cooper M.A., Quiles J.L., Bullón P., Ruiz-Cabello J, & Cordero M.D. (2017). NLRP3-inflammasome inhibition prevents high fat and high sugar diets-induced heart damage through autophagy induction. Oncotarget, 8(59), 99740-99756.

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Western Blot Analysis of Muscle Proteins

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The skeletal muscle tissues were lysed with liquid nitrogen and RIPA lysis buffer (Beyotime, Jiangsu, China). The lysates were homogenized, and the homogenates were centrifuged at 13,000 g for 15 min at 4°C. Proteins were isolated from cultured cells using RIPA lysis buffer and by following the same protocol. The supernatants were collected, and protein concentrations were determined with a BCA Protein Assay kit (Pierce, number 23225). Equal aliquots (30 μg) of the protein samples were applied to 10% SDS-PAGE gels, transferred to polyvinylidene fluoride (PVDF) membranes, and blocked with 5% skim milk TBST (Tris-buffered Saline Tween-20) buffer. The membranes were incubated with primary antibodies including anti-PGC-1, anti-MnSOD, anti-p53, and anti-GAPDH (1 : 1,000; Santa Cruz Biotechnology) as well as anti-Atg7, anti-p62, and anti-LC3 (1 : 1,000; Cell Signaling) at 4°C overnight. Then, the membranes were incubated with anti-rabbit or anti-mouse antibodies at room temperature for 1 h. The protein bands were captured and documented through a CCD system (Image Station 2000MM, Kodak, Rochester, NY, USA) or a gel image analysis system (ChemiDox XRS, Bio-Rad, USA).

Huang Y.F., Lu L., Zhu D.J., Wang M., Yin Y., Chen D.X, & Wei L.B. (2016). Effects of Astragalus Polysaccharides on Dysfunction of Mitochondrial Dynamics Induced by Oxidative Stress. Oxidative Medicine and Cellular Longevity, 2016, 9573291.

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Mitochondrial p38β Phosphorylation Analysis

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Immunoblotting was performed using the following primary antibodies: anti-phospho-p38 (sc-17852-R, Santa Cruz), anti-p38β (sc-6187-R, Santa Cruz), anti-β-actin (sc-130656, Santa Cruz), anti-MnSOD (SC-30080, Santa Cruz), and anti-COX IV (#4844S, Cell Signaling Technology). To detect phosphorylated p38β (p-p38β) in mitochondria, anti-p-p38 antibody was first conjugated to protein A-Sepharose beads to immunoprecipitate all phosphorylated p38 kinase isoforms from mitochondrial lysate. The p-p38 complex was then blotted for p38β to yield p-p38β. Immunoreactive bands were visualized by the enhanced chemiluminescence method (Pierce ECL 2 Western Blotting Substrate) and quantified by NIH ImageJ software.

Luo T., Liu H, & Kim J.K. (2016). Estrogen Protects the Female Heart from Ischemia/Reperfusion Injury through Manganese Superoxide Dismutase Phosphorylation by Mitochondrial p38β at Threonine 79 and Serine 106. PLoS ONE, 11(12), e0167761.

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Murine Macrophage Cell Line Study

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The murine macrophage cell line J774-A1 and the culture Dulbecco’s Modified Eagle’s Medium (DMEM) were obtained from ATCC^® (Manassas, VA, USA). Fetal bovine serum (FBS), Bio-Rad protein assay, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), SP, LPS, H₂O₂ and anti-laminin A/C antibody were obtained from Sigma-Aldrich (Saint Louis, Missouri, USA). Anti-NF-κB, anti-IκBα, anti-COX-2, anti-MnSOD, anti-HO-1, and anti β-actin antibody for Western blot analysis were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Peroxidase-conjugated anti-mouse secondary antibody, peroxidase-conjugated goat and anti-rabbit IgG were obtained from Jackson Immuno Research (West Grove, PA, USA). All compounds used in in vivo study were purchased from Sigma-Aldrich Company Ltd. (Poole, United Kingdom). All solutions used for in vivo infusions were prepared using non-pyrogenic saline (0.9% wt/vol NaCl; Baxter Healthcare Ltd., Thetford, United Kingdom).

Filippone A., Lanza M., Campolo M., Casili G., Paterniti I., Cuzzocrea S, & Esposito E. (2020). The Anti-Inflammatory and Antioxidant Effects of Sodium Propionate. International Journal of Molecular Sciences, 21(8), 3026.

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