Nitrotyrosine Antibody (306507) [Unconjugated]

Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 3 mM peroxynitrite, 3 mM inactivated peroxynitrite, or 100 µM peroxyvanadate for 1 hour and recombinantE. coliDNAK treated with 1 mM 4-hydroxynonenal or 1 mM peroxyynitrite for 1 hour. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Nitrotyrosine Monoclonal Antibody (Catalog # MAB3248), followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF007). The lysates were also probed with Phospho-Tyrosine Monoclonal Antibody (MAB1676). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Nitrotyrosine was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using 25 µg/mL Mouse Anti-Nitrotyrosine Monoclonal Antibody (Catalog # MAB3248) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained (green). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Nitrotyrosine was detected in immersion fixed A549 human lung carcinoma cells using Mouse Anti-Nitrotyrosine Monoclonal Antibody (Catalog # MAB3248) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to Cytoplasmic. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Nitrotyrosine was detected in immersion fixed HepG2 human hepatocellular carcinoma cells using Mouse Anti-Nitrotyrosine Monoclonal Antibody (Catalog # MAB3248) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to Cytoplasmic. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

• Reactivity: All-Multi
• Applications: WB, ICC/IF
• Clone: 306507
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

Nitrotyrosine Antibody (306507) [Unconjugated] Summary

• Immunogen: Nitrotyrosine-modified KLH
• Specificity: Detects Nitrotyrosine adducts on proteins in Western blots. It does not cross-react with phosphotyrosine or 4‑hydroxynonenal adducts. Unfixed cells, tissues, and proteins can be treated with Peroxynitrite (Catalog # AR006) for use as positive controls with this antibody.
• Source: N/A
• Isotype: IgG3
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8-25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in TBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Nitrotyrosine Antibody (306507) [Unconjugated]

• Nitrotyrosine

Background

3-Nitrotyrosine is formed when tyrosine is reacted with peroxynitrite. Since peroxynitrite is formed from nitric oxide and superoxide anion, nitrotyrosine adducts on proteins have been used as markers of oxidative cellular damage and macrophage activation. Elevated nitrotyrosine immunoreactivity has been found in inflammation, osteoarthritis, neurodegenerative diseases, and ischemic damage to the heart and brain.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Nitrotyrosine Antibody (MAB3248)(12)

Publications using MAB3248

• Todorovi?, S;?erani?, MS;Toškovi?, B;Dikli?, M;Mitrovi? Ajti?, O;Suboti?ki, T;Vukoti?, M;Dragojevi?, T;Živkovi?, E;Opri?, S;Stojiljkovic, M;Ga?i?, J;?olakovi?, N;Crnokrak, B;?oki?, VP;?iki?, D; Proinflammatory Microenvironment in Adenocarcinoma Tissue of Colorectal Carcinoma International journal of molecular sciences 2024-09-19 [PMID: 39337548] (Western Blot, Human)
• JH Garcia, EA Akins, S Jain, KJ Wolf, J Zhang, N Choudhary, M Lad, P Shukla, S Gill, W Carson, L Carette, A Zheng, S Kumar, MK Aghi Multi-omic screening of invasive GBM cells in engineered biomaterials and patient biopsies reveals targetable transsulfuration pathway alterations bioRxiv : the preprint server for biology, 2023-02-24;0(0):. 2/24/2023 [PMID: 36865128]
• D ?iki?, A Bogdanovi?, D Markovi?, O Mitrovi?-A, T Suboti?ki, M Dikli?, M Vukoti?, T Dragojevi?, E Živkovi?, JF Santibanez, VP ?oki? Inflammation Promotes Oxidative and Nitrosative Stress in Chronic Myelogenous Leukemia Biomolecules, 2022-02-03;12(2):. 2022-02-03 [PMID: 35204748] (ICC, Human)
• CD Ene, SR Georgescu, M Tampa, C Matei, CI Mitran, MI Mitran, MN Penescu, I Nicolae Cellular Response against Oxidative Stress, a Novel Insight into Lupus Nephritis Pathogenesis Journal of personalized medicine, 2021-07-22;11(8):. 2021-07-22 [PMID: 34442337] (ELISA Capture, Human)
• CD Ene, MN Penescu, SR Georgescu, M Tampa, I Nicolae Posttranslational Modifications Pattern in Clear Cell Renal Cell Carcinoma Metabolites, 2020-12-27;11(1):. 2020-12-27 [PMID: 33375435] (ELISA Capture, Human)
• Prpar Mihevc S, Zako�ek Pipan M, �trbenc M et al. Nitrosative Stress in the Frontal Cortex From Dogs With Canine Cognitive Dysfunction Frontiers in Veterinary Science 11/19/2020 [PMID: 33330694]
• Prpar Mihevc S, Zako�ek Pipan M, �trbenc M et al. Nitrosative Stress in the Frontal Cortex From Dogs With Canine Cognitive Dysfunction Frontiers in Veterinary Science 2020-11-19 [PMID: 33330694]
• Uddin MM, Ibrahim MMH, Aryal D, Briski KP Sex-dimorphic moderate hypoglycemia preconditioning effects on Hippocampal CA1 neuron bio-energetic and anti-oxidant function Mol. Cell. Biochem. 8/10/2020 [PMID: 32779041]
• Alan G Jardine Deleterious effects of phosphate on vascular and endothelial function via disruption to the nitric oxide pathway Nephrol. Dial. Transplant., 2017-10-01;0(0):. 2017-10-01 [PMID: 27448672] (Western Blot, Human)
• Hongyun Li, Kalani Ruberu, Sonia Sanz Muñoz, Andrew M. Jenner, Adena Spiro, Hua Zhao et al. Apolipoprotein D modulates amyloid pathology in APP/PS1 Alzheimer's disease mice Neurobiology of Aging 5/1/2015 [PMID: 25784209]
• Teng Liu, Dan-dan Hou, Qian Zhao, Wei Liu, Pan-pan Zhen, Jian-ping Xu et al. Phytoestrogen alpha -Zearalanol attenuates homocysteine-induced apoptosis in human umbilical vein endothelial cells BioMed Research International 10/1/2013 [PMID: 24195080]
• Zhao Z, Chen Y, Wang J Age-related retinopathy in NRF2-deficient mice. PLoS ONE, 2011-04-29;6(4):e19456. 2011-04-29 [PMID: 21559389] (IHC-Fr, Mouse)

Review for Nitrotyrosine Antibody (MAB3248) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer IHC-P Human 10/11/2017

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: human cornea
• Species: Human
• Lot: XFNO31512B