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Anti 3 nitrotyrosine antibody

Manufactured by Abcam
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The Anti-3-Nitrotyrosine antibody is a laboratory tool used for the detection and quantification of 3-nitrotyrosine, a post-translational modification of proteins. This antibody can be utilized in various immunological techniques, such as Western blotting, ELISA, and immunohistochemistry, to study the presence and levels of 3-nitrotyrosine in biological samples.

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Lab products found in correlation

Antibody against β actin, by Merck Group (1 mentions) Anti rac1, by Merck Group (1 mentions) Phospho akt at ser473, by Cell Signaling Technology (1 mentions) Anti nitrotyrosine antibody, by Merck Group (1 mentions) Tetramethylrhodamine tmrm, by Thermo Fisher Scientific (1 mentions) Gw9662, by Merck Group (1 mentions) Mitotracker, by Thermo Fisher Scientific (1 mentions) Tgf β1, by Merck Group (1 mentions) A1r eclipse ti confocal microscope, by Nikon (1 mentions) Image pro plus 6, by Media Cybernetics (1 mentions) Anti zo 1 antibody, by Thermo Fisher Scientific (1 mentions) Hoechst 33258, by Merck Group (1 mentions) Anti inos antibody, by Abcam (1 mentions) 2 7 dichlorodihydrofluorescein diacetate, by Thermo Fisher Scientific (1 mentions)

4 protocols using anti 3 nitrotyrosine antibody

1

Antibody Immunodetection of Metabolic Proteins

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Antibodies against PPAR-γ, Akt1, phospho-Akt at Ser473 and VDAC were purchased from Cell Signaling (Danvers, MA). Antibodies against eNOS, p67phox, and p47 phox were purchased from BD Transduction laboratories (San Jose, CA). The anti-Rac1 and anti-phospho S617-eNOS were purchased from EMD Millipore (Temecula, CA). The anti-nitrotyrosine antibody was obtained from EMD Millipore (Billerica, MA). The anti-3-Nitrotyrosine antibody (cat#:ab110282) was obtained from Abcam (Cambridge, MA). The anti-CrAT was obtained from Proteintech (Rosemont, IL). The nitro-Tyr350 Akt1–specific antibody was raised against a synthetic peptide antigen CGRLPF(Y-NO2)NQDHEKL, where Y-NO2 represents 3-nitrotyrosine as described [33 (link)]. Antibody against β-actin, TGF- β1 and GW9662 were purchased from Sigma (St. Louis, MO). MitoTracker (cat#: M22425) and Tetramethylrhodamine (TMRM) were obtained from Invitrogen (Carlsbad, CA).
Sun X., Lu Q., Yegambaram M., Kumar S., Qu N., Srivastava A., Wang T., Fineman J.R, & Black S.M. (2020). TGF-β1 attenuates mitochondrial bioenergetics in pulmonary arterial endothelial cells via the disruption of carnitine homeostasis. Redox Biology, 36, 101593.
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2

Quantifying Nitrosative Stress in Rat RCECs

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RCECs were grown on a coated coverslip (3×104 cells per well), allowed to attach overnight and exposed to various experimental conditions. After treatment for 72 h, rat RCECs were fixed in fresh cold 4% paraformaldehyde at 4°C for 30 min and washed three times in PBS. Cells were blocked for 1 h in 5% normal goat serum and incubated with the primary antibody (anti-3-nitrotyrosine antibody 1:100; Abcam, Cambridge, MA, USA) in a moist chamber at 4°C overnight. Following three washes with PBS, the cells were incubated for 1 h at room temperature with a fluorescein-conjugated goat anti-mouse secondary antibody (1:00; Zhong Shan Jin Qiao, Beijing, People’s Republic of China). The coverslips were then mounted onto slides with 4′,6-diamidino-2-phenylindole–containing medium. Images were collected on a Nikon A1R Eclipse Ti confocal microscope (Nikon Corporation) and processed with Image-Pro Plus 6.0 software (Media Cybernetics, Silver Spring, MD, USA).
Fan C., Qiao Y, & Tang M. (2017). Notoginsenoside R1 attenuates high glucose-induced endothelial damage in rat retinal capillary endothelial cells by modulating the intracellular redox state. Drug Design, Development and Therapy, 11, 3343-3354.
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3

Immunofluorescence Analysis of ICH Markers

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Immunofluorescence was performed at 72 h after ICH as previously described [9 (link),32 (link)]. After antigen retrieval by heat treatment, the sections were incubated at 4°C overnight with primary antibodies: anti-iNOS antibody (Abcam, Cambridge, MA, USA), anti-3-Nitrotyrosine antibody (Abcam, Cambridge, MA, USA), anti-ZO-1 antibody (Invitrogen, Carlsbad, CA, USA). It was then incubated with the appropriate fluorescence conjugated secondary antibodies for 1.5 h at room temperature. Nuclei were stained by Hoechst 33258 (Sigma-Aldrich, St. Louis, MO, USA) for 10 min at room temperature. The slices were observed underneath a fluorescence microscope (Bx51, Olympus Corporation, Shinjuku-ku, Japan).
Chen M., Li X., Zhang X., He X., Lai L., Liu Y., Zhu G., Li W., Li H., Fang Q., Wang Z, & Duan C. (2015). The inhibitory effect of mesenchymal stem cell on blood–brain barrier disruption following intracerebral hemorrhage in rats: contribution of TSG-6. Journal of Neuroinflammation, 12, 61.
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4

Assessing Oxidative Stress in Adipose Arterioles

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Reactive oxygen species production was assessed using 5 μM
2’, 7’-dichlorodihydrofluorescein diacetate (Invitrogen, Eugene
OR), a cell-permeable redox-sensitive fluorophore that was previously used to
measure light-chain induced cellular ROS production (Shi et al., 2010 (link)) through conversion to the highly
fluorescent 2’, 7’-dichlorofluorescein (DCF). Living subject
adipose arterioles were exposed for 1 hour to 2 μM Aβ ±
PEGSOD (250 U/mL). At 45 minutes of treatment, acetylcholine
10−4M was administered and DCF was administered after 60
minutes of Aβ treatment. Fluorescence signal was measured using Olympus
IX51 fluorescent microscope (excitation 495, emission 525 nM). Living adipose
arteriole nitrotyrosine content was measured to assess reactive nitrogen
species-induced nitrative stress (Mohiuddin et
al., 2006) using Western blot (anti-3-nitrotyrosine antibody, Abcam,
Cambridge MA) densitometry normalized to β-actin content. Vehicle or
Aβ (2 μM) ± PEGSOD (250 U/mL) was administered to
arterioles for 1 hour and acetylcholine 10−4M was administered
at 45 minutes of treatment.
Truran S., Franco D.A., Roher A.E., Beach T.G., Burciu C., Serrano G., Maarouf C.L., Schwab S., Anderson J., Georges J., Reaven P, & Migrino R.Q. (2014). Adipose and leptomeningeal arteriole endothelial dysfunction induced by β-amyloid peptide: a practical human model to study Alzheimer's disease vasculopathy. Journal of neuroscience methods, 235, 123-129.
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