Anti nitrotyrosine

Manufactured by Merck Group

Sourced in United States, Germany, Italy

Anti-nitrotyrosine is a lab equipment product used for the detection and quantification of nitrotyrosine, a post-translational modification that occurs in proteins under conditions of nitrosative stress. It serves as a marker for oxidative and nitrosative damage.

Automatically generated - may contain errors

Lab products found in correlation

Facsscan, by BD (4 mentions) Anti inos, by BD (3 mentions) Anti caspase 1, by Adipogen (2 mentions) Anti β actin, by Merck Group (2 mentions) Anti ho 1, by Santa Cruz Biotechnology (2 mentions) Cellquest software, by BD (2 mentions) Anti cox2, by BD (2 mentions) Cell quest software version 4, by BD (2 mentions) Anti 8 oxo dg, by Merck Group (1 mentions) Anti nrf2, by Santa Cruz Biotechnology (1 mentions) Anti pten, by Cell Signaling Technology (1 mentions) Anti p akt, by Abcam (1 mentions) Anti ki67, by Abcam (1 mentions) Anti phospho stat3 p stat3, by Cell Signaling Technology (1 mentions) Vectastain elite abc reagent, by Vector Laboratories (1 mentions) Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Lsm800 confocal microscope, by Zeiss (1 mentions) Fluorescent secondary antibody, by Thermo Fisher Scientific (1 mentions) Anti lectin, by Vector Laboratories (1 mentions) Anti neun, by Abcam (1 mentions)

Close spelling variants of this product

Nitrotyrosine (32 citations) Anti-nitrotyrosine antibody (29 citations) Rabbit anti-nitrotyrosine (8 citations) Anti-3-nitrotyrosine (6 citations) Anti-nitrotyrosine rabbit polyclonal antibody (5 citations) Anti-nitrotyrosine (NT) polyclonal antibody (2 citations) Anti-3-nitrotyrosine (3-NT, (2 citations) Mouse anti-nitrotyrosine (2 citations) Primary anti-nitrotyrosine (2 citations) Mouse anti‐nitrotyrosine antibody (2 citations)

27 protocols using anti nitrotyrosine

Immunohistochemical Profiling of Oxidative Stress Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunohistochemical analysis was carried out using a standard avidin–biotin peroxidase complex method. Antibodies used in the study include: anti-8-oxo-dG (mouse monoclonal; JalCA), anti-nitrotyrosine (rabbit polyclonal; Millipore), anti-COX-2 (M-19; goat polyclonal; Santa Cruz), anti-Nrf2 (rabbit polyclonal; Santa Cruz), anti-Ki-67 (rabbit polyclonal, Abcam), anti-p-AKT (rabbit polyclonal; Abcam), and anti-PTEN (rabbit monoclonal; Cell Signaling Technology). Sections were then counterstained with hematoxylin. The total number of cells, the number of positively stained cells (including 1+, 2+, and 3+), and the immunostaining intensity were quantified using the Aperio ScanScope GL system (Vista, CA).

Chen J.X., Li G., Wang H., Liu A., Lee M.J., Reuhl K., Suh N., Bosland M.C, & Yang C.S. (2015). Dietary tocopherols inhibit PhIP-induced prostate carcinogenesis in CYP1A-humanized mice. Cancer letters, 371(1), 71-78.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Colon Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunohistochemical analysis was carried out using 4 μm paraffin-embedded sections of mouse colon. A standard avidin–biotin peroxidase complex method was employed. Briefly, deparaffinized sections on glass slides were heated in a microwave oven for antigen retrieval, endogenous peroxidase was quenched using 3% H₂O₂, and tissue sections were incubated at 4°C overnight with primary antibodies. Biotinylated secondary antibody and Vectastain Elite ABC reagent (Vector Laboratories, Burlingame, CA) were applied with 3, 3′-diaminobenzidine. Sections were then counterstained with hematoxylin. Primary antibodies used in the study included: anti-8-oxo-deoxyguanosine (8-oxo-dG) (mouse monoclonal; JalCA), anti-nitrotyrosine (rabbit polyclonal; Millipore), anti-Nuclear Factor Kappa B p65 subunit (NF-κB p65) (rabbit polyclonal, Abcam), anti-phospho-STAT3 (p-STAT3) (rabbit monoclonal, Cell Signaling Technology), anti-phospho-Histone H2AX (γH2AX) (rabbit monoclonal, Cell Signaling Technology), and anti-cleaved caspase-3 (rabbit polyclonal, Cell Signaling Technology). The number of positively stained cells and total number of cells were quantified using the Aperio ScanScope GL system (Vista, CA). About 10,000–30,000 cells from colon tumors or adjacent tissues were analyzed per mouse.

Chen J.X., Liu A., Lee M.J., Wang H., Yu S., Chi E., Reuhl K., Suh N, & Yang C.S. (2016). δ- and γ-Tocopherols Inhibit PhIP/DSS-induced Colon Carcinogenesis by Protection against Early Cellular and DNA Damages. Molecular carcinogenesis, 56(1), 172-183.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Mouse Brain

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mice were anesthetized and immediately perfused with PBS and 4% paraformaldehyde (PFA) as previously described [32 (link)]. The brain was post-fixed in 4% PFA overnight and transferred to 30% sucrose in PBS. 50 μm thick sections were prepared by vibratome. Then, the brain sections were incubated with primary antibodies as following: anti-Caspase-1 (1:500, AdipoGen, CA, USA), anti-NeuN (1:300, Abcam, Cambridge, UK), anti-Lectin (1:300, DL-1177, Vector Laboratories, USA), anti-Nitrotyrosine (1:1000, Millipore, CA, USA) for overnight at 4 °C. After being rinsed three times, the sections were incubated with appropriate fluorescent secondary antibodies (Invitrogen, Carlsbad, CA) for 2 h at room temperature. Finally, the sections were stained with DAPI (0.5 μg/ml) for 5 min and mounted in Vectashield medium (Vector Laboratories, CA). Immunofluorescence images were performed with a Zeiss LSM800 confocal microscope (Carl Zeiss, Germany). The 3D surface visualization of grayscale intensity was plotted through ImageJ v1.50H and its “Interactive 3D Surface Plot” plug-in.

Sun M., Mao X.F., Li Z.M., Zhu Z.H., Gong D.M., Lu L., Chen X., Zhang Y., Fukunaga K., Ji Y., Gu A.H., Lu Y.M, & Han F. (2021). Endothelial peroxynitrite causes disturbance of neuronal oscillations by targeting caspase-1 in the arcuate nucleus. Redox Biology, 47, 102147.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Lung Tissue

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

At the end of the experiments, slices of lung tissue were incubated with anti-ICAM-1 (1/100 in PBS, SCB), anti-P-selectin (1/100 in PBS, SCB), anti-CD-8 (1:450 in PBS, sc-7970), anti-CD-11b (1:450 in PBS, sc-1186), anti-CD18 (1:350 in PBS, sc-18862), anti-CD4 (1:350 in PBS, sc-13573), anti-PAR (1/100 in PBS, SCB), and anti-nitrotyrosine (1:200 in PBS Millipore) as previously described [64 (link),69 (link),82 (link),83 (link),84 (link)]. After that, sections were treated with peroxidase-conjugated goat anti-rabbit IgG or bovine anti-mouse IgG secondary antibodies (1:2000 Jackson Immuno Research, West Grove, PA, USA). A biotin-conjugated goat anti-rabbit IgG or the biotin-conjugated goat anti-mouse IgG and avidin-biotin peroxidase combination (Vector Laboratories, Burlingame, CA, USA) were used to identify specific markers. Using an imaging device (Leica DM6, Milan, Italy), immunohistochemical pictures were captured (LasX Navigator, Milan, Italy). The number of positive pixels were analyzed with Image J plug-in [64 (link),69 (link),82 (link),83 (link),84 (link)]. All immunohistochemical analyses were carried out by two observers blind to the treatment [67 (link),68 (link),85 (link),86 (link),87 (link)].

Genovese T., Duranti A., Monaco F., Siracusa R., Fusco R., Impellizzeri D., D’Amico R., Cordaro M., Cuzzocrea S, & Di Paola R. (2023). Inhibition of Fatty Acid Amide Hydrolase (FAAH) Regulates NF-kb Pathways Reducing Bleomycin-Induced Chronic Lung Inflammation and Pulmonary Fibrosis. International Journal of Molecular Sciences, 24(12), 10125.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Inflammatory Markers

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunohistochemical examination was performed as previously described [31 (link)], 4 days after DNBS administration. The sections were incubated overnight with the following primary antibodies: anti-CD4⁺ (Santa Cruz Biotechnology (SCB), 1:100 in PBS, v/v, sc-19641, D.B.A, Milan, Italy); anti-CD8⁺ (SCB, 1:100 in PBS, v/v, sc-1177, D.B.A, Milan, Italy); anti-ICAM-1 (SCB, 1:100 in PBS, v/v, sc-107, D.B.A, Milan, Italy), anti-P-selectin (1:100 in PBS, v/v, sc-8419 SCB, D.B.A, Milan, Italy), anti-PARP (1:100 in PBS, v/v, sc-8007 SCB, D.B.A, Milan, Italy), and anti-nitrotyrosine (1:200 in PBS, v/v, Millipore, D.B.A, Milan, Italy). All slices were washed with PBS and then treated as previously reported [32 (link)]. Briefly, slices were incubated with universal secondary antibody. Specific labeling was detected with a biotin- conjugated goat anti-rabbit IgG and avidin-biotin peroxidase complex (Vector).
Five stained sections from each mouse were scored in a blinded fashion and observed using a Leica DM6 microscope (Leica Microsystems SpA, Milan, Italy) following a typical procedure [33 (link)]. The histogram profile is related to the positive pixel intensity value obtained.

Impellizzeri D., Fusco R., Genovese T., Cordaro M., D’Amico R., Trovato Salinaro A., Ontario M.L., Modafferi S., Cuzzocrea S., Di Paola R., Calabrese V, & Siracusa R. (2022). Coriolus Versicolor Downregulates TLR4/NF-κB Signaling Cascade in Dinitrobenzenesulfonic Acid-Treated Mice: A Possible Mechanism for the Anti-Colitis Effect. Antioxidants, 11(2), 406.

+ Open protocol

+ Expand

Protein Expression Analysis in Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

NTS tissues were added with pre-cooled lysate and then cracked by ultrasonography. The specimens were statically placed on ice for 30 min to be fully cracked and then centrifuged at 12,000 rpm for 15 min at 4°CC. Supernatant was collected for protein assay. The Bradford assay (Generay Biotechnology, Shanghai) was used to determine the concentration of protein in tissue. An amount of 5 μg of protein was loaded in each lane. The protein was separated by electrophoresis, transferred onto polyvinylidene fluoride membranes, which were blocked with 0.1% Tween-20 tris-buffered saline containing 10% nonfat milk for 1 h at room temperature, and then incubated overnight at 4°C with the rabbit polyclonal antibodies anti-AT1R (1:800; ab18801, Abcam, USA), anti-ATRAP (1:500; sc-134652, Santa Cruz Biotechnology), anti-Nox2 (1:1,000; Abcam), anti-p67phox (1:1,000; EPITOMICS, USA), and anti-nitrotyrosine (1:1,000; Millipore), followed by the peroxidase-conjugated goat anti-rabbit secondary antibody (1:2,000; SA00001-2, Proteintech Biotechnology). Western blotting reagents (Millipore Corp., Billerica, MA) were used to detect the signal and blots were exposed to X-ray film for densitometric analysis.

Feng X., Guo Q., Xue H., Duan X., Jin S, & Wu Y. (2020). Hydrogen Sulfide Attenuated Angiotensin II-Induced Sympathetic Excitation in Offspring of Renovascular Hypertensive Rats. Frontiers in Pharmacology, 11, 565726.

+ Open protocol

+ Expand

Immunofluorescence Analysis of Retinal Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

Eyes were embedded in Tissue Freezing Medium (Electron Microscopy Sciences) and 10μm thick sections were collected in-round on 12 slides such that each slide contained a representation of all areas of the eye/retina. Slides were rinsed with phosphate buffered saline (PBS) and incubated at room temperature in normal donkey serum at 1:20 in 0.1 M phosphate buffer with 0.5% bovine serum albumin and 0.1% Triton X 100 (PBT) for 2 hours. The slides were incubated overnight at 4°C in anti-nitrotyrosine (1:500, Millipore, Billerica, MA), anti-glial fibrillary acidic protein (GFAP, 1:400, DAKO, Carpinteria, CA), or anti-H ferritin (1:100, Abcam, Cambridge, MA) in PBT, rinsed with PBS and incubated with a secondary antibody (Life Technologies, Grand Island, NY) for 2 hours at room temperature. Slides were rinsed with PBS and mounted in Vectashield Mounting medium with 4’,6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Burlingame, CA) for imaging on a Nikon Eclipse epifluorescence microscope (Nikon, Melville, NY).

Bricker-Anthony C., D'Surney L., Lunn B., Hines-Beard J., Jo M., Bernardo-Colon A, & Rex T.S. (2017). Erythropoietin either Prevents or Exacerbates Retinal Damage from Eye Trauma Depending on Treatment Timing. Optometry and vision science : official publication of the American Academy of Optometry, 94(1), 20-32.

+ Open protocol

+ Expand

Evaluating Oxidative Stress in Cell and Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

To examine ROS in tissue culture, B16.SIY cells were seeded in 100 mm culture dishes, incubated overnight and treated with cephalexin (50 μM) for 1 hour prior to 6 or 12 Gy. N-acetyl-cysteine (NAC, 3 mM) was added 1 hour prior to cephalexin as indicated. After 24 hours, cells were washed with PBS and trypsinized. CellROX Deep Red reagent (0.75 μM, Thermo) was added to suspended cells for 60 minutes at 37°C in the dark. Viability stain Sytox Blue (1 μM, Thermo) was added for the last 15 minutes. Stained cells were analyzed with a BD Fortessa flow cytometer and FlowJo software. Dead cells and debris were excluded.
To evaluate ROS in tumors, 5 μm sections from FFPE were blocked in 1% BSA + 5% normal horse serum, probed overnight at 4°C with anti-nitrotyrosine (5 μg/ml, Millipore), anti-8-OHdG (1/200, Abcam) or anti-4-HNE (1/200, Abcam), and detected with secondary-HRP conjugates and DAB (Vector Laboratories). Sections were mounted under PolyMount (Polysciences) and brightfield images were collected using an Axiovert 40 microscope.

Labay E., Mauceri H.J., Efimova E.V., Flor A.C., Sutton H.G., Kron S.J, & Weichselbaum R.R. (2016). Repurposing cephalosporin antibiotics as pro-senescent radiosensitizers. Oncotarget, 7(23), 33919-33933.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Nitrotyrosine

Check if the same lab product or an alternative is used in the 5 most similar protocols

For immunohistochemical analysis, antigen retrieval was performed by heating sections in 10 mM sodium citrate buffer (pH 6.0) for 20 min. Endogenous peroxidase was quenched by incubating the sections with Peroxidase Suppressor (Thermo Scientific, Rockford, IL, USA) for 15 min at RT. The slides were blocked with Non-Serum Protein Block (Dako, Carpinteria, CA, USA) for 20 min at RT. Primary antibodies were prepared in antibody diluent solution (0.5% non-fat dry milk and 1% BSA in TBS) and incubated overnight at 4 °C. The concentration of primary antibody and dilution for Anti-Nitrotyrosine was 1:6000 (Millipore, Temecula, CA, USA). The specificity of the nitrotyrosine antibody binding was confirmed by blocking the antibody with 3-nitrotyrosine (10 mM). Immunoreactivity was detected by Dako Envision+ System-HRP (Dako, Carpinteria, CA, USA).

Alund A.W., Mercer K.E., Pulliam C.F., Suva L.J., Chen J.R., Badger T.M, & Ronis M.J. (2016). PARTIAL PROTECTION BY DIETARY ANTIOXIDANTS AGAINST ETHANOL-INDUCED OSTEOPENIA AND CHANGES IN BONE MORPHOLOGY IN FEMALE MICE. Alcoholism, clinical and experimental research, 41(1), 46-56.

+ Open protocol

+ Expand

Assessing Lipid Peroxidation and Nitrotyrosine

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lipid peroxidation was assessed by the malondialdehyde (MDA) and the amounts of lipid peroxides were detected by high performance liquid chromatography and were expressed as μmol MDA/mg protein [89 (link)]. Nitrotyrosine was measured by Western blot with anti-nitrotyrosine (Millipore, 05-233, Darmstadt, Germany) 1/1000 dilution, and a secondary anti-mouse antibody (lot. Number OI192080; 1/5000; Pierce Biotechnology, Rockford, IL, USA). The signals obtained on immunoblot determinations were scanned and quantified by densitometric analysis with a chemoluminescence detection device (Odyssey Imaging System, Li-Cor Biosciences, Lincoln, NE, USA).

Herrera E.A., Farías J.G., González-Candia A., Short S.E., Carrasco-Pozo C, & Castillo R.L. (2015). Ω3 Supplementation and Intermittent Hypobaric Hypoxia Induce Cardioprotection Enhancing Antioxidant Mechanisms in Adult Rats. Marine Drugs, 13(2), 838-860.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!