Oxidized PTP Active Site Antibody (335636) [Unconjugated]

Western blot shows oxidized PTP immunoprecipitated from lysates of HepG2 human hepatocellular carcinoma cell line using Rat Anti-SHP-2 antibody. PVDF membrane was probed with 1 µg/mL Mouse Anti-Oxidized PTP Active Site Monoclonal Antibody (Catalog # MAB2844) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band for Oxidized SHP-2 was detected at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Topoisomerase inhibitors induce JAK2-STAT1-CXCL1 and migration through ROS. a Relative DCFH-DA levels in SW480 and SW620 cells treated with VP-16 (V, 20 μM), ADM (A, 0.2 μg/ml), or CPT-11 (C, 80 μg/ml) for 0.5 h. P, a positive control with Rosup H2O2 (50 μg/ml, 0.5 h). (B) Western blot of oxidized PTPs after treatement with VP-16 (20 μM), ADM (0.2 μg/ml), or CPT-11 (80 μg/ml) for 0.5 h. c Confirmation of VP-16-induced PTP1B oxidization. After treatment with VP-16 (20 μM, 0.5 h), cell lysates (200 μg per sample) were immunoprecipitated with 1 μg anti-PTP1B or pre-immune IgG for 12 h. Precipitates and cell lysates (input, 50 μg per sample) were analyzed by Western blot with anti-oxPTP and anti-PTP1B. d Western blot of JAK2 and STAT1 phosphorylation and CXCL1 expression in cells pretreated with GSH (10 mM) for 2 h and subsequently treated with VP-16 (20 μM) or CPT-11 (80 μg/ml) for 0.5 h. e Migration assay of cells treated with GSH (10 mM) plus VP-16 (20 μM) or CPT-11 for 24 h Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31438997), licensed under a CC-BY license. Not internally tested by R&D Systems.

Topoisomerase inhibitors induce JAK2-STAT1-CXCL1 and migration through ROS. a Relative DCFH-DA levels in SW480 and SW620 cells treated with VP-16 (V, 20 μM), ADM (A, 0.2 μg/ml), or CPT-11 (C, 80 μg/ml) for 0.5 h. P, a positive control with Rosup H2O2 (50 μg/ml, 0.5 h). (B) Western blot of oxidized PTPs after treatement with VP-16 (20 μM), ADM (0.2 μg/ml), or CPT-11 (80 μg/ml) for 0.5 h. c Confirmation of VP-16-induced PTP1B oxidization. After treatment with VP-16 (20 μM, 0.5 h), cell lysates (200 μg per sample) were immunoprecipitated with 1 μg anti-PTP1B or pre-immune IgG for 12 h. Precipitates and cell lysates (input, 50 μg per sample) were analyzed by Western blot with anti-oxPTP and anti-PTP1B. d Western blot of JAK2 and STAT1 phosphorylation and CXCL1 expression in cells pretreated with GSH (10 mM) for 2 h and subsequently treated with VP-16 (20 μM) or CPT-11 (80 μg/ml) for 0.5 h. e Migration assay of cells treated with GSH (10 mM) plus VP-16 (20 μM) or CPT-11 for 24 h Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31438997), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: All-Multi
• Applications: WB
• Clone: 335636
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

Oxidized PTP Active Site Antibody (335636) [Unconjugated] Summary

• Immunogen: KLH-coupled perfomic acid oxidized synthetic peptide
  VHCSAG
• Specificity: Detects Oxidized PTP Active Site. Will react with any oxidized phosphatase containing the consensus sequence. Detects oxidized but not unoxidized DEP-1, PTP1B, TC-PTP, and SHP-2. Other phosphatases have not been tested.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • 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 PBS with Trehalose. *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.

Notes

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

Alternate Names for Oxidized PTP Active Site Antibody (335636) [Unconjugated]

• Oxidized PTP Active Site

Background

Oxidation of the essential cysteine in the active site of protein tyrosine phosphatases (PTPs) by exogenous or intracellular oxidants such as hydrogen peroxide is believed to play an important role in regulating the activity of these enzymes. The initial oxidation to a sulfenic acid (-SOH) is reversible, but the oxidation often progresses to an irreversible sulfonic acid (-SO₃H) that can be detected by modification-specific antibodies. The active sites of PTPs contain the consensus amino acid sequence (V/I)HCSXG, a sequence essential for catalytic activity.

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 Oxidized PTP Active Site Antibody (MAB2844)(18)

Publications using MAB2844

• Chen, Y;Lin, Y;Hong, J;Wang, J;Li, B;Liu, Z;Ma, Y;Sun, X;Wang, S;Li, M;Wu, M;Fu, D;Li, J;Wang, M;Hu, L; Oroxylin A reverses SHP-2 oxidative inactivation in GPVI signaling to suppress platelet activation and thrombus formation Thrombosis journal 2025-03-18 [PMID: 40102927] (Western Blot, Human)
• Cappabianca, L;Ruggieri, M;Sebastiano, M;Sbaffone, M;Martelli, I;Ruggeri, P;Di Padova, M;Farina, AR;Mackay, AR; Molecular Characterization and Inhibition of a Novel Stress-Induced Mitochondrial Protecting Role for Misfolded TrkAIII in Human SH-SY5Y Neuroblastoma Cells International journal of molecular sciences 2024-05-17 [PMID: 38791513] (Western Blot, Human)
• Seeyoung Choi, Jan Lee, Teri Hatzihristidis, Guillaume Gaud, Avik Dutta, Awadhesh Arya et al. THEMIS increases TCR signaling in CD4 + CD8 + thymocytes by inhibiting the activity of the tyrosine phosphatase SHP1 Science Signaling 5/9/2023 [PMID: 37159521]
• NC Luong, Y Abiko, T Shibata, K Uchida, E Warabi, M Suzuki, T Noguchi, A Matsuzawa, Y Kumagai Redox cycling of 9,10-phenanthrenequinone activates epidermal growth factor receptor signaling through S-oxidation of protein tyrosine phosphatase 1B J Toxicol Sci, 2020-01-01;45(6):349-363. 2020-01-01 [PMID: 32493877] (Neutralization, Western Blot, Human)
• Jiafei Liu, Like Qu, Lin Meng, Chengchao Shou Topoisomerase inhibitors promote cancer cell motility via ROS-mediated activation of JAK2-STAT1-CXCL1 pathway Journal of Experimental & Clinical Cancer Research 12/1/2019 [PMID: 31438997]
• R Duval, LC Bui, C Mathieu, Q Nian, J Berthelet, X Xu, I Haddad, J Vinh, JM Dupret, F Busi, F Guidez, C Chomienne, F Rodrigues- Benzoquinone, a leukemogenic metabolite of benzene, catalytically inhibits the protein tyrosine phosphatase PTPN2 and alters STAT1 signaling J. Biol. Chem., 2019-06-27;0(0):. 2019-06-27 [PMID: 31248982] (Western Blot)
• Marcus Grohmann, Florian Wiede, Garron T. Dodd, Esteban N. Gurzov, Geraldine J. Ooi, Tariq Butt et al. Obesity Drives STAT-1-Dependent NASH and STAT-3-Dependent HCC Cell 11/15/2018 [PMID: 30454647]
• KB Neves, AC Montezano, R Alves-Lope, T Bruder-Nas, RM Costa, RS Costa, RM Touyz, RC Tostes Upregulation of Nrf2 and Decreased Redox Signaling Contribute to Renoprotective Effects of Chemerin Receptor Blockade in Diabetic Mice Int J Mol Sci, 2018-08-19;19(8):. 2018-08-19 [PMID: 30126255] (Western Blot, Mouse)
• JH Kim, SJ Park, U Chae, J Seong, HS Lee, SR Lee, S Lee, DS Lee Peroxiredoxin 2 mediates insulin sensitivity of skeletal muscles through regulation of protein tyrosine phosphatase oxidation Int. J. Biochem. Cell Biol., 2018-03-29;99(0):80-90. 2018-03-29 [PMID: 29605633] (Western Blot, Mouse)
• Seeyoung Choi, Paul E. Love Detection of Intracellular Reduced (Catalytically Active) SHP-1 and Analyses of Catalytically Inactive SHP-1 after Oxidation by Pervanadate or H2O2 BIO-PROTOCOL 1/5/2018 [PMID: 29552589]
• S Torii, C Kubota, N Saito, A Kawano, N Hou, M Kobayashi, R Torii, M Hosaka, T Kitamura, T Takeuchi, H Gomi The pseudophosphatase phogrin enables glucose-stimulated insulin signaling in pancreatic ?-cells J. Biol. Chem., 2018-02-26;0(0):. 2018-02-26 [PMID: 29483197] (Western Blot, Mouse)
• W Wu, AJ Hale, S Lemeer, J den Hertog Differential oxidation of protein-tyrosine phosphatases during zebrafish caudal fin regeneration Sci Rep, 2017-08-16;7(1):8460. 2017-08-16 [PMID: 28814789] (Immunoprecipitation, Zebrafish)
• S Choi, C Warzecha, E Zvezdova, J Lee, J Argenty, R Lesourne, L Aravind, PE Love THEMIS enhances TCR signaling and enables positive selection by selective inhibition of the phosphatase SHP-1 Nat. Immunol, 2017-02-27;0(0):. 2017-02-27 [PMID: 28250424] (Western Blot, Human)
• Jiefei Tong, Mohamed Helmy, Florence M. G. Cavalli, Lily Jin, Jonathan St-Germain, Robert Karisch et al. Integrated analysis of proteome, phosphotyrosine-proteome, tyrosine-kinome, and tyrosine-phosphatome in acute myeloid leukemia PROTEOMICS 5/5/2017 [PMID: 28176486]
• Hirakawa S, Saito R, Ohara H Dual oxidase 1 induced by Th2 cytokines promotes STAT6 phosphorylation via oxidative inactivation of protein tyrosine phosphatase 1B in human epidermal keratinocytes. J. Immunol., 2011-03-16;186(8):4762-70. 2011-03-16 [PMID: 21411736] (Western Blot, Human)
• Sharma P, Chakraborty R, Wang L, Min B, Tremblay ML, Kawahara T, Lambeth JD, Haque SJ Redox regulation of interleukin-4 signaling. Immunity, 2008-10-17;29(4):551-64. 2008-10-17 [PMID: 18957266] (Western Blot, Human)
• Chen YY, Chu HM, Pan KT, Teng CH, Wang DL, Wang AH, Khoo KH, Meng TC Cysteine S-nitrosylation protects protein-tyrosine phosphatase 1B against oxidation-induced permanent inactivation. J. Biol. Chem., 2008-10-07;283(50):35265-72. 2008-10-07 [PMID: 18840608] (Western Blot, Human)