Horseradish peroxidase hrp

Manufactured by Merck Group

Sourced in United States, Germany, United Kingdom, China, Japan, Spain, Australia

Horseradish peroxidase (HRP) is an enzyme derived from the horseradish plant. It is a widely used laboratory reagent known for its ability to catalyze oxidation-reduction reactions in the presence of hydrogen peroxide. HRP is commonly employed as a reporter molecule in various analytical techniques, including immunoassays, Western blotting, and histochemical staining.

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Lab products found in correlation

213 protocols using horseradish peroxidase hrp

Western Blot Analysis of Cell Signaling

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Cell lysates were prepared in RIPA buffer (Beyotime) containing protease inhibitors and were separated by SDS polyacrylamide gel electrophoresis (SDS-PAGE), and transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, Billerica, MA, USA). Incubation with primary antibodies recognizing p16 (CST, USA), p21 (CST, USA), pRb (CST, USA), GNAQ (Abcam, USA), PLK2 (CST, USA), tubulin, and β-actin (Santa Cruz, USA) was followed by incubation with the appropriate secondary antibodies conjugated to horseradish peroxidase (HRP) (Millipore, Billerica, MA, USA). Signals were developed with enhanced chemiluminescence.

Wu W.P., Zhou M.Y., Liu D.L., Min X., Shao T., Xu Z.Y., Jing X., Cai M.Y., Xu S., Liang X., Mo M., Liu X, & Xiong X.D. (2021). circGNAQ, a circular RNA enriched in vascular endothelium, inhibits endothelial cell senescence and atherosclerosis progression. Molecular Therapy. Nucleic Acids, 26, 374-387.

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Quantification of Smad4 and β-catenin Interactions

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Cells were washed with PBS, harvested, and lysed in 0.2% NP-40 lysis buffer. The protein lysates were subjected to immunoprecipitation with antibodies containing anti-Smad4 or anti-β-catenin (Abcam, MA) overnight, and then incubated with protein A/G Plus Agarose (Fisher) for 4 hours. The immunocomplex was washed with PBS three times, followed by mixture with SDS buffer. For western blotting, whole cell or co-precipitated extracts were separated by a Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE), blotted onto Immobilon polyvinyl difluoride (PVDF) membrane (Millipore, MA), and incubated with primary antibodies containing anti-Trb3, anti-Smad4, anti-β-catenin or anti-GAPDH. The membranes were subsequently incubated in secondary antibodies conjugated with Horseradish Peroxidase (HRP) (Millipore, MA) and visualized with chemiluminescent HRP (Denville Scientific, NJ).

Fan J., Lee C.S., Kim S., Zhang X., Pi-Anfruns J., Guo M., Chen C., Rahnama M., Li J., Wu B.M., Aghaloo T.L, & Lee M. (2020). Trb3 Controls Mesenchymal Stem Cell Lineage Fate and Enhances Bone Regeneration by Scaffold-Mediated Local Gene Delivery. Biomaterials, 264, 120445.

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Western Blot Analysis of Phosphotyrosine Proteins

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Proteins separated by SDS-PAGE were transferred by semi-dry electroblotting onto polyvinylidene difluoride membranes in triplicate to detect proteins with three different antibodies. Phosphotyrosine protein detection was carried out with primary antibodies: the membrane blots were incubated overnight at 4°C using a 1:1000 dilution of mouse monoclonal IgG anti-phosphotyrosine 4G10 antibody (Millipore). Detection of tagged protein with His₆ or GST was carried out with primary antibodies using a 1:1000 dilution of mouse monoclonal IgG_2ak anti-histidine tagged antibody (Millipore) or mouse monoclonal IgG anti-GST tag antibody (Millipore), respectively. All membranes were incubated for 1 h with 1:20000 dilution of secondary antibody (goat anti mouse IgG) coupled with horseradish peroxidase (HRP) (Millipore). Supersignal substrate (Thermo scientific) was used for HRP detection and visualised by exposure to Amersham Hyperfilm ECL (GE).

Kang H.J., Gilbert C., Badeaux F., Atlan D, & LaPointe G. (2015). A tyrosine phosphorylation switch controls the interaction between the transmembrane modulator protein Wzd and the tyrosine kinase Wze of Lactobacillus rhamnosus. BMC Microbiology, 15, 40.

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Synthesis and Characterization of Multifunctional Hydrogels

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All reagents were used as received unless otherwise stated. N-isopropylacrylamide, methacrylic acid (MAA), azobisisobutyronitrile (AIBN), diethyl ether (Et2O), bovine serum albumin (BSA), fluorescein isothiocyanate (FITC), rhodamine isothiocyanate (RITC), hexamethylenediamine, N-[(1R,8S,9s)-bicyclo[6.1.0]non-4-yn-9-ylmethyloxycarbonyl]-1,8-diamino-3,6-dioxaoctane (BCN-NH2), 11-azido-3,6,9-trioxaundecan-1-amine (N3-NH2), bis-N-succinimidyl polyethylene glycol (NHS-PEG35), deuterated chloroform (CDCl3) (99.8 atom %), 4,7,10, 13,16,19,22,25,32,35,38,41,44,47,50,53hexadecaoxa-28,29-dithiahexapentacontanedioic acid di-N-succinimidyl ester (NHS-PEG16-DS), 2-ethyl-1-hexanol, glutathione (GSH), dithiothreitol (DTT), dihydroxyacetophenone (DHAP), diammonium hydrogen citrate (DAHC), trifluoroacetic acid (TFA), glucose oxidase (GOx) (252100 U/mg), horseradish peroxidase (HRP) (172.2 U/mg), α-glucosidase (ALG,19.3 U/mg) and maltose were purchased from Sigma-Adrich. Acetonitrile, Amplex Red, DyLight 405 and Hoechst 33258 were purchased from Thermo Fisher Scientific. Hexanes and glucose was purchased from VWR. N-ethyl-N'-(3dimethylaminopropyl) carbodiimide hydrochloride (EDAC) from Alfa Aesar, and N-[(9H-fluoren-9-ylmethoxy)carbonyl]-Lalanyl-L-alanine (Fmoc-Ala-Ala-OH) was purchased from Bachem. Dialysis bags with MWCO 1,000 or 12,000-14,000 Da were purchased from Millipore.

Gobbo P., Patil A.J., Li M., Harniman R., Briscoe W.H, & Mann S. (2018). Programmed assembly of synthetic protocells into thermoresponsive prototissues. Nature materials, 17(12).

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Quantifying Aflatoxins in Food Samples

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STG, aflatoxins (B1, B2, G1, G2 and M1), acetonitrile (ACN), methanol (MeOH), acetone, chloroform, n-heptane, sulphuric acid, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, sodium carbonate, sodium bicarbonate, sodium chloride, magnesium sulphate, Tween 20, bovine serum albumin (BSA), keyhole limpet hemocyanin (KLH), horseradish peroxidase (HRP), sodium borohydride and dimethylformamide (DMF) were purchased from Sigma-Aldrich (Dorset, UK & Zwijndrecht, the Netherlands). Primary-secondary amine (PSA) bulk sorbent was purchased from Agilent (Amstelveen, the Netherlands). Substrate 3,3',5,5'-tetramethylbenzidine (TMB) for HRP enzyme was obtained from Neogen (Lansing, USA).
Samples were blended using IKA A11 Basic laboratory mill (IKA, Staufen, Germany) and centrifuged in Sigma 4K10 centrifuge (Sigma, Osterode am Harz, Germany). Microtiter plates were read on BioTek EL808 type ELISA plate reader (BioTek, Bad Friedrichshall, Germany).

Oplatowska-Stachowiak M., Reiring C., Sajic N., Haasnoot W., Brabet C., Campbell K., Elliott C.T, & Salden M. (2018). Development and in-house validation of a rapid and simple to use ELISA for the detection and measurement of the mycotoxin sterigmatocystin. Analytical and bioanalytical chemistry, 410(12).

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Characterization of Pharmacological Agents

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1-(3-Chlorophenyl)piperazine hydrochloride (mCPP), 1-[5-(thiophen-2-ylmethoxy)-1H-indol-3-yl]propan-2-amine (BW 723C86), 1-[3-(2-dimethylaminoethyl)-1H-indol-5-yl]-N-methylmethanesulfonamide succinate (sumatriptan), Nω-Nitro-L-arginine methyl ester hydrochloride (LNAME), Evans Blue and horseradish peroxidase (HRP) were purchased from Sigma-Aldrich GmbH (Germany). 4-(6-ethoxy-1-methoxythioxanthen-9-ylidene)-1-methyl piperidine maleate (BF-1) was synthetized by Biofrontera Bioscience GmbH at Pharm-Eco Laboratories, Inc. Devens (USA). Saline (0.9%) was used as vehicle for mCPP, L-NAME, HRP and Evans Blue. DMSO in a maximal concentration of 0.1% diluted in saline was used as vehicle for BW 723C86, sumatriptan and BF-1. DMSO (0.1% in saline) by itself did not induce PPE (data not shown). Evans Blue was diluted in saline and filtered with 0.2 μm filter. All test compounds were freshly prepared for each experiment. All doses are expressed per kg bodyweight (BW).

Hunfeld A., Segelcke D., Bäcker I., Mecheri B., Hemmer K., Dlugosch E., Andriske M., Paris F., Zhu X, & Lübbert H. (2015). Hypoxia facilitates neurogenic dural plasma protein extravasation in mice: a novel animal model for migraine pathophysiology. Scientific Reports, 5, 17845.

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Antioxidant Capacity Assay Protocol

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Gallic acid and Folin-Ciocalteau were obtained from Fluka Chemical Co. (Buchs, Switzerland) and propylene glycol from Chemco LTDA. 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,4,6-tris(2-pyridyl-s-triazine (TPTZ), o-dianisidine dihydrochloride, ethylene glycol bis (-aminoethyl ether)-N,N,N0,N0-tetraacetic acid (EGTA), reduced glutathione (GSH), 5,5-dithio-bis-(2-nitrobenzoic acid) (DTNB), hexadecyltrimethylammonium bromide (HTAB), luminol, horseradish peroxidase (HRP), xanthine, xanthine oxidase (XOD) and Trolox were obtained from Sigma Chemical Co. (St. Louis, MO, USA). Raw materials for formulations were obtained from Galena (Campinas, SP, Brazil) . All other reagents used were of pharmaceutical grade.

Takayama K.S., Monteiro M.C., Saito P., Pinto I.C., Nakano C.T., Martinez R.M., Thomaz D.V., Verri WA J.r., Baracat M.M., Arakawa N.S., Russo H.M., Zeraik M.L., Casagrande R., Couto R.O.D, & Georgetti S.R. (2022). Rosmarinus officinalis extract-loaded emulgel prevents UVB irradiation damage to the skin. Anais da Academia Brasileira de Ciencias, 94(4).

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Coniferyl Aldehyde Oxidation Assay

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Chemicals including coniferyl aldehyde (4-hydroxy-3-methoxycinnamaldehyde, Sigma-Aldrich), horseradish peroxidase (HRP, 250-330 units/mg, Sigma-Aldrich), hydrogen peroxide solution (30% wt in H 2 O, Sigma-Aldrich), sodium borohydride (Sigma-Aldrich) and ethyl acetate ([99.5%, Sigma-Aldrich) with analytical grades were used.

Aminzadeh S., Zhang L, & Henriksson G. (2017). A possible explanation for the structural inhomogeneity of lignin in LCC networks. Wood Sci Technol., 51(6).

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Antioxidant Assays of Natural Extracts

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Agarose, Au(III) chloride trihydrate (HAuCl₄·3H₂O), gallic acid, luminol, sucrose, fluorescein sodium salt, 2,2′-azobis (2-amidinopropane) dihydrochloride (AAPH), and horseradish peroxidase (HRP) were purchased from Sigma Aldrich (St. Louis, MO). All other chemicals were of the highest analytical grade.
Teas, herbal infusions, and aloe antioxidant drink (containing 30% of Aloe vera juice and pulp) are commercially available products and were purchased from a local store. Extra virgin olive oil (EVOO) samples and EVOO waste product extracts (oleuropein-enriched extracts from olive tree leaves and hydroxytyrosol-enriched extracts from olive mill wastewaters) were obtained from Italian manufacturing companies participating in the VIOLIN (Valorization of Italian OLive products through INnovative analytical tools) research project funded by Cariplo Foundation within the “Agroalimentare e Ricerca” (AGER) program.

Calabria D., Guardigli M., Severi P., Trozzi I., Pace A., Cinti S., Zangheri M, & Mirasoli M. (2021). A Smartphone-Based Chemosensor to Evaluate Antioxidants in Agri-Food Matrices by In Situ AuNP Formation. Sensors (Basel, Switzerland), 21(16), 5432.

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Pluronic Micelles for Antioxidant Assays

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Pluronic F127 ((PEO)₂₀₀(PPO)₆₅(PEO)₂₀₀, Mw 12600), Pluronic P104 ((PEO)₅₄(PPO)₆₃(PEO)₅₄, Mw 5900), Pluronic P85 ((PEO)₅₂(PPO)₄₀(PEO)₅₂, Mw 4600) and Pluronic F88 ((PEO)₂₀₈(PPO)₃₉(PEO)₂₀₈, Mw 11400), were supplied by BASF corporation. Hydrogen tetrachloroaurate (III) trihydrate (HAuCl₄·3H₂O), PEO (Mw 8000), sodium chloride (NaCl), hydrochloric acid (HCl), sodium hydroxide (NaOH), 4-acetamidophenol (AAP), horseradish peroxidase (HRP), and diethylenetriaminepentaacetic acid (DTPA) were purchased from Sigma-Aldrich, St. Louis, MO. 1-Hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine (CMH), EPR buffer, diethyldithiocarbamic acid sodium salt (DETC), and deferoxamine (DF) were purchased from Noxygen Science Transfer and Diagnostics, Elzach, Germany All reagents were used as received without farther purification. All aqueous solutions were prepared using bi-distilled water (Millipore, Billerica, MA, USA).

Sokolsky-Papkov M, & Kabanov A. (2019). Synthesis of Well-Defined Gold Nanoparticles Using Pluronic: The Role of Radicals and Surfactants in Nanoparticles Formation. Polymers, 11(10), 1553.

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