Anti ho 1 antibody

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The Anti-HO-1 antibody is a laboratory reagent used for the detection and analysis of the Heme Oxygenase 1 (HO-1) protein. HO-1 is an enzyme that plays a crucial role in the degradation of heme, a component of hemoglobin. This antibody can be used in various immunological techniques, such as Western blotting, immunohistochemistry, and ELISA, to identify and quantify the presence of HO-1 in biological samples.

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Anti-HO-1 (75 citations) HO-1 antibody (5 citations) Anti-heme oxygenase 1 (HO-1) antibody (2 citations)

7 protocols using anti ho 1 antibody

Immunoblotting Analysis of Endothelial Factors

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The following antibodies were used: anti-eNOS antibody, anti-p-eNOS antibody, anti-VASP antibody and anti-p-VASP antibody were purchased from Cell Signaling Technology. Anti-CSE antibody, anti-GPx antibody, anti-HO-1 antibody and anti-GAPDH antibody were purchased from Santa Cruze.
All drugs were purchased from Sigma-aldrich unless otherwise stated.

Wu D., Hu Q., Xiong Y., Zhu D., Mao Y, & Zhu Y.Z. (2017). Novel H2S-NO hybrid molecule (ZYZ-803) promoted synergistic effects against heart failure. Redox Biology, 15, 243-252.

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Flow Cytometric Analysis of Antioxidant Enzymes

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C6 cells (5.0 × 10⁴/well), primary astrocytes and mixed glial cells (3.5 × 10⁴/well) were seeded on 96-well plate and treated, after 24 h, with IS (15–60 μM). After 24 h cells were collected, washed with PBS and then incubated in Fixing Solution for 20 min and then incubated in Fix Perm Solution for 30 min, at 4°C. Anti-HO-1 antibody (Santa Cruz Biotechnologies; sc-10789; 1:100), NQO1 antibody (Santa Cruz Biotechnologies; sc-376023; 1:100), superoxide dismutase (SOD) antibody (Santa Cruz Biotechnologies; sc-11407; 1:100), anti-iNOS (BD Transducion Laboratories; 610431; 1:100) antibody, anti-COX-2 (BD Transducion Laboratories; 610203; 1:100) antibody and anti-nitrotyrosine antibody (Millipore; 06-284; 1:100) were added to C6 cells, primary astrocytes and mixed glial cells. The secondary antibody (Immuno Reagents; used at diluition 1:100) was added in Fix Perm Solution and cells were evaluated using a fluorescence-activated cell sorting (FACSscan; Becton Dickinson) and elaborated with Cell Quest software as previously reported (Adesso et al., 2013 (link)).

Adesso S., Magnus T., Cuzzocrea S., Campolo M., Rissiek B., Paciello O., Autore G., Pinto A, & Marzocco S. (2017). Indoxyl Sulfate Affects Glial Function Increasing Oxidative Stress and Neuroinflammation in Chronic Kidney Disease: Interaction between Astrocytes and Microglia. Frontiers in Pharmacology, 8, 370.

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Antibody Characterization for Cell Signaling Studies

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The antibodies were purchased from the following manufacturers: anti-GAPDH antibody (Santa Cruz Biotechnology: sc-25778), anti-PGRMC1 antibody for western blotting (Cell Signaling: 13856S), anti-PGRMC1 antibody for immunofluorescent staining (Abcam: ab48012), anti-PPARγ antibody (Abcam: ab59256), anti-FABP4 antibody (Abcam: ab66682), anti-LDL-R antibody (R&D: AF2255), anti-VLDL-R antibody (R&D: AF2258), anti-Tf-R antibody (Abcam: ab84039), anti-Na-K ATPase α1 antibody (Abcam: ab7671), anti-GLUT1 antibody (Abcam: ab115730), anti-GLUT4 antibody (Abcam: ab33780), anti-Akt antibody (Cell Signaling: #9272S), anti-pAkt antibody (Cell Signaling: #4060S), and anti-HO-1 antibody (Santa Cruz Biotechnology: sc-136960).

Furuhata R., Kabe Y., Kanai A., Sugiura Y., Tsugawa H., Sugiyama E., Hirai M., Yamamoto T., Koike I., Yoshikawa N., Tanaka H., Koseki M., Nakae J., Matsumoto M., Nakamura M, & Suematsu M. (2020). Progesterone receptor membrane associated component 1 enhances obesity progression in mice by facilitating lipid accumulation in adipocytes. Communications Biology, 3, 479.

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Triptolide, Sulforaphane, and Antioxidant Pathways

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Triptolide (>99% purity) was purchased from DND Pharm-Technology Co. (Shanghai, China). 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) was purchased from MP Biomedicals (Santa Ana, CA, USA). Sulforaphane (SFN, ≥98% purity) was purchased from Enzo Life Sciences (Lausen, Switzerland). Anti-NQO1 antibody was purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-Nrf2 antibody, anti-GCLC antibody and anti-HO-1 antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-Histone H3 antibody and anti-β-actin antibody were purchased from Cell Signaling Technology (Beverly, MA, USA). Other chemicals were of analytical grade from commercial suppliers.

Li J., Shen F., Guan C., Wang W., Sun X., Fu X., Huang M., Jin J, & Huang Z. (2014). Activation of Nrf2 Protects against Triptolide-Induced Hepatotoxicity. PLoS ONE, 9(7), e100685.

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AMPK and HO-1 Modulation in RAW 264.7 Cells

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RAW 264.7 cells were cultured in Rosewell Park Memorial Institute-1640 Medium supplemented with 10% fetal bovine serum, penicillin, and 100 μg/ml streptomycin (Gibco BRL Gaithersburg, MD). Antibodies against AMPK, p-AMPK, acetyl-CoA carboxylase (ACC), p-ACC, p38, and phospho-p38 (p-p38) were purchased from Cell Signaling Technology (Beverly, MA). Anti-HMGB1 antibody was purchased from Abcam (Cambridge, MA). Anti-HO-1 antibody, anti-β-actin antibody, scrambled siRNAs, and siAMPKα1 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). The enhanced chemiluminescence western blotting detection reagent was purchased from Amersham (Buckinghamshire, UK). The p38 MAPK inhibitors (PD98059, SB203580, and SP600125) were purchased from Calbiochem (San Diego, CA). All other chemicals, including LPS (Escherichia coli 0111:B4), naringin, zinc protoporphyrin (ZnPP; an inhibitor of HO-1 enzyme), and MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) were purchased from Sigma-Aldrich (St. Louis, MO). LPS and inhibitors were treated on cultured cells with appropriate concentration: 1 μg/ml of LPS, 10 μM of PD989052, SB203580, SP600125, and ZnPP.

Gil M., Kim Y.K., Hong S.B, & Lee K.J. (2016). Naringin Decreases TNF-α and HMGB1 Release from LPS-Stimulated Macrophages and Improves Survival in a CLP-Induced Sepsis Mice. PLoS ONE, 11(10), e0164186.

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Western Blot Analysis of HO-1

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Cells were lysed using a method as previously described[25 (link)]. Proteins of lysed cells were then separated by SDS-PAGE on 10% polyacrylamide gels. Separated proteins were transferred to nitrocellulose membranes for western blotting using a standard protocol as described previously[26 (link)]. A rabbit polyclonal anti-HO-1 antibody was purchased from Santa Cruz Biotechnology. SDS PAGE gels were stained using a Coomassie Blue staining kit (Bio-Rad). Molecular weight markers are based on the mobility of All Blue Protein Standard (Bio-Rad), ranging from 10 kDa to 250 kDa. The gel was imaged with an Odyssey Scanner (LI-COR Bioscience, Lincoln NE).

Mu J., Zhuang X., Wang Q., Jiang H., Deng Z.B., Wang B., Zhang L., Kakar S., Jun Y., Miller D, & Zhang H.G. (2014). Interspecies communication between plant and mouse gut host cells through edible plant derived exosome-like nanoparticles. Molecular nutrition & food research, 58(7), 1561-1573.

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Evaluation of Wy14,643 and Fibrates

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Wy14,643 (pirinixic acid, 4-Chloro-6-(2,3-xylidino)-2-pyrimidinylthioacetic acid) and fenofibrate were from Sigma (St. Louis, MO) and ICN Pharmaceuticals, Inc. (Montréal, Canada), respectively. Gemfibrozil was from Pfizer Canada (Kirkland, Canada). Fetal bovine serum (FBS) and other cell culture reagents were from Invitrogen (Burlington, Canada). The SYBR Green PCR mix was purchased from Applied Biosystems (Foster City, CA). Protein assay reagents were obtained from Bio-Rad Laboratories Inc. (Marnes-la-Coquette, France). The anti-HO-1 antibody was from Santa Cruz (Santa Cruz, CA). The anti-actin antibody was purchased from Sigma and the anti-rabbit IgG antibody was from GE Healthcare (Piscataway Township, NJ).

Bigo C., Kaeding J., El Husseini D., Rudkowska I., Verreault M., Vohl M.C, & Barbier O. (2014). PPARα: A Master Regulator of Bilirubin Homeostasis. PPAR Research, 2014, 747014.

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