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Anti catalase

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Anti-catalase is a laboratory reagent used to inhibit the activity of the enzyme catalase. Catalase is a common enzyme found in most living organisms that catalyzes the decomposition of hydrogen peroxide into water and oxygen. Anti-catalase can be used to study the role of catalase in biological systems.

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Lab products found in correlation

Anti pparα, by Thermo Fisher Scientific (3 mentions) Anti cyp2e1, by Abcam (3 mentions) Anti nadsyn, by Antibodies-online (3 mentions) Anti tdo2, by Thermo Fisher Scientific (3 mentions) Anti nmnat1, by LifeSpan BioSciences (3 mentions) Anti β actin, by Merck Group (3 mentions) Anti chop, by Cell Signaling Technology (2 mentions) Anti p22phox, by Santa Cruz Biotechnology (1 mentions) Anti nf kb, by Santa Cruz Biotechnology (1 mentions) Anti ve cadherin, by Cell Signaling Technology (1 mentions) Anti sod1, by Santa Cruz Biotechnology (1 mentions) Anti inos, by Santa Cruz Biotechnology (1 mentions) Anti zo 1, by Thermo Fisher Scientific (1 mentions) D glucose, by Merck Group (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Ab51254, by Abcam (1 mentions) Ab125068, by Abcam (1 mentions) Anti β actin, by Novus Biologicals (1 mentions) A8529, by ABclonal (1 mentions) Arg10112, by Arigo Biolaboratories (1 mentions)

Spelling variants of this product

Catalase (805 citations) Rabbit anti-catalase (CAT) (2 citations)

15 protocols using anti catalase

1

Erucin Synthesis and Cell Assays

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Erucin was produced by myrosinase-catalyzed hydrolysis of glucoerucin isolated from Eruca sativa Mill. defatted seed meals according to Citi et al. [21 (link)]. Erucin was dissolved in dimethyl sulfoxide (DMSO; Merck KGaA, Darmstadt, Germany) and this solution (10−2 M) was freshly diluted in the appropriate culture medium.
An aqueous solution of d-(+)-Glucose (45% w/w; Merck KGaA, Darmstadt, Germany) was diluted before each experiment in culture medium up to a final concentration of 25 mM. CelLytic™ MT Cell Lysis Reagent, Fluoromount Aqueous Mounting Medium, and 3 kDa FITC-Dextran were obtained from Life Technologies (Carlsbad, CA, USA). Anti-VE-Cadherin was obtained from Cell Signalling (Danvers, MA, USA) and anti-ZO-1 was obtained from Life Technologies (Carlsbad, CA, USA). Anti-SOD-1, anti-iNOS, anti-NF-KB and anti-p22phox antibodies were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-catalase and anti-β-actin were obtained from Merck KGaA, (Darmstadt, Germany).
Martelli A., Piragine E., Gorica E., Citi V., Testai L., Pagnotta E., Lazzeri L., Pecchioni N., Ciccone V., Montanaro R., Di Cesare Mannelli L., Ghelardini C., Brancaleone V., Morbidelli L, & Calderone V. (2021). The H2S-Donor Erucin Exhibits Protective Effects against Vascular Inflammation in Human Endothelial and Smooth Muscle Cells. Antioxidants, 10(6), 961.
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2

Immunoblotting and Immunohistochemistry of Liver Samples

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Paraffin-embedded liver sections were handled using standard methods as described previously [27 ]. The following antibodies were used for immunoblotting: anti-PPARα (Thermo Fisher Scientific), anti-4-HNE (Northwest Life Science Specialties, Vancouver, WA), anti-CHOP (Cell Signaling Technology, Danvers, MA), anti-catalase (EMD Millipore, Burlington, MA), anti-CYP2E1 (Abcam, Cambridge, MA), anti-NADSYN (Antibodies-online, Limerick, PA), anti-TDO2 (Thermo Fisher Scientific), and anti-NMNAT1 (Lifespan Biosciences, Seattle, WA). Methyl green was used to counterstain cell nuclei for 4-HNE staining.
Yue R., Chen G.Y., Xie G., Hao L., Guo W., Sun X., Jia W., Zhang Q., Zhou Z, & Zhong W. (2021). Activation of PPARα-catalase pathway reverses alcoholic liver injury via upregulating NAD synthesis and accelerating alcohol clearance. Free radical biology & medicine, 174, 249-263.
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3

Comprehensive Western Blot Analysis

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We utilized the following antibodies to perform Western blot analysis. The primary antibodies were anti-4-HNE (ARG23717, Arigo Biolaboratories, HSZ, Taiwan), anti-xCT (26864–1-AP, Proteintech, Rosemont, IL, USA), anti-GPX4 (ARG41400, Arigo Biolaboratories, HSZ, Taiwan), anti-GAPDH (ARG10112, Arigo Biolaboratories), anti-β-actin (NB600-501, Novus Biologicals, TPE, Taiwan), anti-catalase (219010, Merck, DA, Germany), anti-SOD2 (# 06–984, Merck), anti-SOD1 (ab51254, Abcam, EN, UK), anti-ULK1 (A8529, ABclonal, Woburn, MA, USA), anti-ND-1 (A5250, ABclonal), anti-UQCRC2 (A4181, ABclonal), anti-p62 (66184–1-Ig, Proteintech), and anti-Lamp2 (ab125068, Abcam). The secondary antibodies were anti-mouse IgG (7076, Cell Signaling, Danvers, MA, USA) and anti-rabbit IgG (7074, Cell Signaling).
Bow Y.D., Ko C.C., Chang W.T., Chou S.Y., Hung C.T., Huang J.L., Tseng C.H., Chen Y.L., Li R.N, & Chiu C.C. (2023). A novel quinoline derivative, DFIQ, sensitizes NSCLC cells to ferroptosis by promoting oxidative stress accompanied by autophagic dysfunction and mitochondrial damage. Cancer Cell International, 23, 171.
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4

Western Blot Analysis of Hepatic Proteins

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Mouse livers homogenates were loaded onto 12% sodium dodecyl sulfate-polyacrylamide gel, transblotted onto polyvinylidene difluoride membranes (Bio-Rad, Hercules, CA), blocked with 5% nonfat dry milk, and then incubated with the following antibodies, including anti-PPARα, anti-TDO2 (Thermo Fisher Scientific), anti-catalase (EMD Millipore), anti-CYP2E1, anti-ADH, anti-ALDH2, anti-NQO1 (Abcam), anti-GSTM1 (Proteintech, Rosemont, IL), anti-ATF4, anti-GAPDH (Cell Signaling Technology), anti-NADSYN (Antibodies-online), anti-NMNAT1 (Lifespan Biosciences) and anti-β-actin (Sigma-Aldrich, St. Louis, MO). The membranes were then incubated with HRP-conjugated goat anti-rabbit IgG, goat anti-mouse IgG, or rabbit anti-goat IgG (Thermo Fisher Scientific). The bound complexes were detected by enhanced chemiluminescence and quantified by densitometry analysis.
Yue R., Chen G.Y., Xie G., Hao L., Guo W., Sun X., Jia W., Zhang Q., Zhou Z, & Zhong W. (2021). Activation of PPARα-catalase pathway reverses alcoholic liver injury via upregulating NAD synthesis and accelerating alcohol clearance. Free radical biology & medicine, 174, 249-263.
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5

Protein Detection Reagents for Western Blotting and Immunofluorescence

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Detecting reagents used for western blotting include: streptavidin-HRP (Cell Signaling, 3999S, 1:3000), anti-Myc (Cell Signaling, 2276S, 1:3000), anti-V5 (Abcam, ab27671, 1:3000), anti-β actin (GenScript, A00702, 1:3,000), anti-FLAG (GNI, GNI14110-FG, 1:3000), anti-ABCD3 (Sigma, P0497, 1:3000), anti-MFN1 (Cell Signaling, D6E2S, 1:3000), anti-MFN2 (Cell Signaling, D1E9, 1:3000). Antibodies used for immunofluorescence include: anti-MFN1 (Cell Signaling, D6E2S, 1:500), anti-MFN2 (Cell Signaling, D1E9, 1:500), anti-calnexin (Abcam, ab22595, 1:500), anti-EEA1 (Cell Signaling, C45B10, 1:500), anti-GM130 (Abcam, ab52649, 1:500), anti-LAMP1 (Cell Signaling, D2D11, 1:500), anti-ABCD3 (Sigma, SAB4200181, 1:500), anti-ABCD3 (Sigma, P0497, 1:500), anti-catalase (Merck/Millipore, 219010, 1:500), anti-FLAG (GNI, GNI14110-FG, 1:500), anti-FLAG (Proteintech, 20543-1-AP, 1:500), anti-Myc (Cell Signaling, 2276S, 1:500), anti-COX4 (Proteintech, 11242-1-AP, 1:500).
Huo Y., Sun W., Shi T., Gao S, & Zhuang M. (2022). The MFN1 and MFN2 mitofusins promote clustering between mitochondria and peroxisomes. Communications Biology, 5, 423.
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6

Assessing PPARα and Catalase in Liver

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Liver nuclear extracts were prepared using a nuclear extraction kit from Cayman Chemical. PPARα activity in the extracts was assessed by detecting double-stranded DNA-bound PPARα with specific antibodies in an ELISA format (Cayman Chemical). Catalase activity in whole liver homogenates was examined using a catalase activity assay kit (BioVision). The following antibodies were used for immunoblotting: anti-PPARα (Thermo Fisher Scientific), anti-4-HNE (Northwest Life Science Specialties, Vancouver, WA), anti-CHOP (Cell Signaling Technology, Danvers, MA), anti-catalase (EMD Millipore, Burlington, MA), anti-CYP2E1 (Abcam, Cambridge, MA), anti-NADSYN (Antibodies-online, Limerick, PA), anti-TDO2 (Thermo Fisher Scientific), and anti-NMNAT1 (Lifespan Biosciences, Seattle, WA). Methyl green was used to counterstain cell nuclei for 4-HNE staining.
Yue R., Chen G.Y., Xie G., Hao L., Guo W., Sun X., Jia W., Zhang Q., Zhou Z, & Zhong W. (2021). Activation of PPARα-catalase pathway reverses alcoholic liver injury via upregulating NAD synthesis and accelerating alcohol clearance. Free radical biology & medicine, 174, 249-263.
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7

Transcriptional and Translational Regulation in HAEC

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RNA isolation, reverse transcription and SYBR® green-based (Applied Biosystems) quantitative PCR was carried according to standard protocols using a Quant Studio 7 Flex Real Time PCR thermocycler (Applied Biosystems) with the associated sequence detection system and software. Expression was calculated using the ΔΔCT method. Relative gene expression was normalized to β-actin (house-keeping gene). Western blot analyses of HAEC lysates were conducted according to standard protocols using the following specific antibodies:
anti-Sirt3 (rabbit monoclonal, Cell Signalling Technology), anti-C/EBP-β [C19] (rabbit polyclonal, Santa Cruz Biotechnology), anti-SOD2 (rabbit polyclonal, Abcam), anti-catalase (mouse monoclonal, Sigma), anti-glutathion peroxidase 1 (rabbit polyclonal, Novus Biologicals), anti-total eNOS (mouse polyclonal, BD Transduction Laboratories), anti-eNOS (pThr495) (mouse, monoclonal, BD Transduction Laboratories), anti-eNOS (pSer1177) (mouse, monoclonal, BD Transduction laboratories), anti-xanthine oxidase (rabbit polyclonal, Biorbit), anti-NADPH oxidase subunit p22phox (rabbit polyclonal, Biorbit), and anti-β actin (mouse monoclonal, Sigma-Aldrich). Specific signals were detected using species-specific secondary antibodies.
Winnik S., Gaul D.S., Siciliani G., Lohmann C., Pasterk L., Calatayud N., Weber J., Eriksson U., Auwerx J., van Tits L.J., Lüscher T.F, & Matter C.M. (2016). Mild endothelial dysfunction in Sirt3 knockout mice fed a high-cholesterol diet: protective role of a novel C/EBP-β-dependent feedback regulation of SOD2. Basic Research in Cardiology, 111, 33.
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8

Immunohistochemical Analysis of Renal Inflammation and Redox Status

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Immunohistochemical staining was performed on renal sections as described previously [19 (link)]. Infiltration of lymphocytes and macrophages was assayed by staining with anti-CD3 antibody (1:200 dilution; Thermo Fisher Scientific) and anti-Iba1 antibody (1:800 dilution; Abcam, Cambridge, MA, USA), respectively. The number of CD3-positive and Iba1-positive cells was calculated in 20 high power fields (HPFs).
Levels of redox-related enzymes were also assayed by immunohistochemical staining using the following antibodies: anti-catalase (1:500; Sigma-Aldrich); anti-GPX1 (1:300) and anti-iNOS (1:200) (both Abcam). The kidney slides were stained using a biotin-free immunoenzymatic antigen detection system (Expose mouse/rabbit specific HRP/DAB IHC kit, Abcam, USA). The intensity of immunohistochemical staining signal for CAT, GPX1 and inducible nitric oxide synthase (iNOS) was evaluated using the scales described previously [20 (link)], and the positive cells were counted in 20 HPFs.
Li Y., Yan M., Yang J., Raman I., Du Y., Min S., Fang X., Mohan C, & Li Q.Z. (2014). Glutathione S-transferase Mu 2-transduced mesenchymal stem cells ameliorated anti-glomerular basement membrane antibody-induced glomerulonephritis by inhibiting oxidation and inflammation. Stem Cell Research & Therapy, 5(1), 19.
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9

Protein Expression Analysis in Ocular Cells

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Mouse eyecups or ARPE-19 Cells were lysed in RIPA buffer with protease inhibitor cocktail, PMSF and sodium orthovanadate (Santa Cruz Biotechnology) and sonicated by ultrasound (on ice). Protein concentration was quantified using BCA kit (Pierce Biotechnology, Inc., Rockford, IL, United States). Twenty-five micrograms of protein were resolved by SDS-PAGE gel and electro-transferred to nitrocellular membranes. After blocking with 5% non-fat milk, membranes were blotted overnight at 4°C with following primary antibodies: anti-Nrf2 (1:1000; Santa Cruz Biotechnology), anti-XBP1 (1:500; Santa Cruz Biotechnology), anti-Catalase (1:2000; Sigma-Aldrich), anti-SOD1 (1:2000; Abcam), anti-SOD2 (1:1000; Assay Designs, MI, United States) and anti-β-actin (1:5,000; Abcam). After incubation with HRP-conjugated secondary antibodies, membranes were developed with chemiluminescence substrate (Thermo Fisher Scientific, Rockford, IL, United States: #34076) using Vision Works LS image acquisition and analysis software (UVP, Upland, CA, United States), and bands were semi-quantified by densitometry.
Chen C., Zhong Y., Wang J.J., Yu Q., Plafker K., Plafker S, & Zhang S.X. (2018). Regulation of Nrf2 by X Box-Binding Protein 1 in Retinal Pigment Epithelium. Frontiers in Genetics, 9, 658.
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10

Protein Quantification by Western Blot

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Proteins were fractionated by SDS-PAGE and transferred onto a nitrocellulose membrane (Schleicher & Schuell Bioscience). The blot was blocked (1 hour at room temperature) with TBST (Tris-buffered saline, 0.1% Tween-20) containing 5% nonfat milk and then probed with the primary antibodies, in the following dilutions: 1:500 anti-TXNRD2 (Atlas Antibodies), 1:1000 anti-TXNRD2 (clone 1C4 as described earlier24 (link)), 1:10 000 anti-ACTIN (Sigma), 1:5000 anti-catalase (Sigma), 1:500 anti-LC3B (Sigma), 1:2000 anti-HSP60 (BD Bioscience), 1:5000 anti-HSP25 (Biomol GmbH), 1:500 anti–hypoxia inducible factor 1, α subunit (HIF1α) (Santa Cruz Biotechnology,), and 1:1000 anti-p62/SQSTM1 (Sigma). After washing with TBST (2×15 minutes), immunodetection of the signals was performed by using horseradish peroxidase–conjugated, corresponding secondary antibodies followed by 2 additional washing steps (2×10 minutes). Finally, the signal was detected by using ECL Plus Detection Reagent (Amersham Biosciences). Protein expression was quantified by using ImageJ software (National Institutes of Health).
Kiermayer C., Northrup E., Schrewe A., Walch A., de Angelis M.H., Schoensiegel F., Zischka H., Prehn C., Adamski J., Bekeredjian R., Ivandic B., Kupatt C, & Brielmeier M. (2015). Heart-Specific Knockout of the Mitochondrial Thioredoxin Reductase (Txnrd2) Induces Metabolic and Contractile Dysfunction in the Aging Myocardium. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 4(7), e002153.
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