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Ang 2

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Ang-2 is a 498 amino acid secreted glycoprotein that acts as an antagonist of the Tie2 receptor. It inhibits angiogenesis and promotes vascular regression and endothelial cell apoptosis.

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Lab products found in correlation

Ang 1, by R&D Systems (12 mentions) Vascular endothelial growth factor (vegf), by R&D Systems (3 mentions) Pdgf bb, by R&D Systems (2 mentions) Ang 2, by Merck Group (2 mentions) Hepatocyte growth factor (hgf), by R&D Systems (2 mentions) Fgf19, by R&D Systems (2 mentions) Interleukin 6 (il 6), by R&D Systems (2 mentions) G csf, by R&D Systems (2 mentions) Hank s balanced salt solution, by Thermo Fisher Scientific (1 mentions) Coomassie protein assay kit, by Thermo Fisher Scientific (1 mentions) Mip 2, by R&D Systems (1 mentions) Neubauer hemocytometer, by Hausser Scientific (1 mentions) Haosmc, by Lonza (1 mentions) Smgm 2 medium, by Lonza (1 mentions) Collagenase 2, by Worthington (1 mentions) Tumor necrosis factor α tnf α, by R&D Systems (1 mentions) Mouse mfg e8, by R&D Systems (1 mentions) Human mfg e8, by R&D Systems (1 mentions) Hek293 cell, by American Type Culture Collection (1 mentions) Fibroblast growth factor (fgf), by R&D Systems (1 mentions)

Spelling variants of this product

Human Ang-2 DuoSet ELISA kit (2 citations) Human biotinylated ANG-2 (2 citations)

39 protocols using ang 2

1

Angiopoietin-2 Modulation in Transplantation

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The mice (n = 175) were randomly divided into three groups according to Ang2 treatment (0 ng/kg Ang2, control: n = 56; 50 ng/kg Ang2: n = 60; and 500 ng/kg Ang2: n = 59). The mice were given IP injections with 50 or 500 ng/kg Ang2 (R&D System; Minneapolis, MN) at 18 hours and 30 minutes before the transplantation procedures because the half-life of Ang2 was 18 hours [23 (link)]. The first Ang2 injection (before 18 hours) was administered to act on the OT grafts after transplantation; and the second injection (before 30 minutes), to prolong the Ang2 effect during neovascularization. For the control group, normal saline was also injected intraperitoneally by using the same scheme described earlier.
Youm H.W., Lee J., Kim E.J., Kong H.S., Lee J.R., Suh C.S, & Kim S.H. (2016). Effects of Angiopoietin-2 on Transplanted Mouse Ovarian Tissue. PLoS ONE, 11(11), e0166782.
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2

Bronchoalveolar Lavage Cell Analysis

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Immediately after collection of BAL fluids, erythrocytes were lysed using 0.2% saline and the remaining cells were resuspended in Hanks' Balanced Salt Solution (Invitrogen, Grand Island, NY). Total cell count of each BAL sample was determined using a Neubauer hemocytometer (Hausser Scientific, Horsham, PA). Differential cell counts were performed (200 cells for each experimental condition) on cytospin slides stained with Protocol HEMA3 solution (Fisher scientific, Fair Lawn, NJ). MPO levels were determined using Suzuki’s protocol [17 (link)]. Total protein concentration in BAL fluid was measured using Coomassie protein assay kit (Thermo Scientific, Rockford, IL). Ang-2, KC and MIP-2 concentration in plasma, tissue homogenates and BAL fluid was measured using specific ELISAs (Ang-2, R&D Systems, Minneapolis, MN; KC and MIP-2, Millipore Inc., Billerica, MA) according to the manufacturer’s instructions.
Lee J.Y., Linge H.M., Ochani K., Lin K, & Miller E.J. (2016). N-Ethylmaleimide Sensitive Factor (NSF) Inhibition Prevents Vascular Instability following Gram-Positive Pulmonary Challenge. PLoS ONE, 11(6), e0157837.
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3

Isolation and Characterization of Mouse Vascular Smooth Muscle Cells

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Mouse VSMCs were enzymatically isolated as described in the literature [33 (link)]. In brief, the thoracic aortas of the young and aged mice were isolated, and the adventitia and intima were removed from the vessels. The aortae were cut into 1–2-mm pieces and placed into tubes containing 2 mg/mL collagenase II (Worthington Biochemical, Lakewood, NJ, USA) at 37 °C for 3–5 h. The isolated cells were washed with and plated in complete Dulbecco’s Modified Eagle’s Medium (DMEM). To ensure the purity of the isolated cells, > 95% of the cells had to be positive for the two specific smooth muscle cell markers: smooth muscle α-actin and smooth muscle myosin heavy chain for them to be used [34 ]. Early passage VSMCs were treated with Ang II (1 μM, R&D Systems), tumor necrosis factor-α (TNF-α, 20 ng/mL, R&D Systems), or mouse MFG-E8 (250 ng/mL, R&D Systems) for 24 h prior to the subsequent experiments. Human aortic smooth muscle cells (hAoSMCs) were purchased from Lonza (Basel, Switzerland) and cultured in SmGM-2 medium (Lonza) for 24 h in either the presence or absence of human MFG-E8 (250 ng/mL, R&D systems).
Chiang H.Y., Chu P.H, & Lee T.H. (2019). MFG-E8 mediates arterial aging by promoting the proinflammatory phenotype of vascular smooth muscle cells. Journal of Biomedical Science, 26, 61.
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4

Characterization of Primary PASMCs

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Human embryonic kidney (HEK) 293 cells were obtained from American Type Culture Collection (ATCC, Manassas, USA) and maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS). Human pulmonary arterial smooth muscle cells (PASMCs) were purchased from Sciencell (San Diego, CA, USA) and cultured in smooth muscle cell medium (SMCM) as previously described [40 (link)]. Rat PASMCs were isolated and cultured in DMEM supplemented with 10% FBS as reported by Zeng et.al. [15 (link)]. PASMCs at passage 3 or 4 were used for experiments, and the contractile phenotype was confirmed by the determination of contractile marker genes expression, such as a-SMA and SM22 [16 (link)]. Growth factors including PDGFBB, AngII, TGF-b, IGF, VEGF, ET-1, PDGFAA and FGF were obtained from R&D Systems. For the growth factors treatment, PASMCs were starved for 12h in starvation condition (0.2% FBS) and then stimulated with the growth factors at concentration as indicated.
Qian Z., Zhang L., Chen J., Li Y., Kang K., Qu J., Wang Z., Zhai Y., Li L, & Gou D. (2016). MiR-328 targeting PIM-1 inhibits proliferation and migration of pulmonary arterial smooth muscle cells in PDGFBB signaling pathway. Oncotarget, 7(34), 54998-55011.
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5

Macrophage-T cell Crosstalk in Angiotensin II-mediated Inflammation

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Macrophages and CD4+ TCs isolated as described above were divided into the following groups: (1) macrophages, (2) macrophages+Ang II (100 nmol/l, Sigma), (3) macrophages+CD4+ TCs+AngII, and (4) macrophages+CD4+ TCs+Ang II+anti-mouse IL-18-neutralizing monoclonal antibody (anti-IL-18 nAb; 100 ng/ml, R&D Systems). The numbers of macrophages and CD4+ TCs in each group were 5 × 105 and 2.5 × 106, respectively. After treatment for 24 hours in complete DMEM, the iNOS, IFN-γ, and IL-6 mRNA levels in the macrophages were detected by qRT-PCR. In addition, SMCs were treated with the supernatant of the cultures described above and Ang II for 12 hours, and the Bax and Bcl2 mRNA levels in the SMCs were measured.
Hu H., Zhang G., Hu H., Liu W., Liu J., Xin S., Zhao X., Han L., Duan L., Huang X, & Chang C. (2019). Interleukin-18 Expression Increases in the Aorta and Plasma of Patients with Acute Aortic Dissection. Mediators of Inflammation, 2019, 8691294.
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6

Hippocampal RAS Enzyme and Peptide Quantification

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Rats were sacrificed after urethane anesthesia (1.2 g/kg i.p., Sigma-Aldrich). Animals were perfused with 4°C saline, and hippocampal tissues were quickly removed and placed in freeze storage at −70°C. Thawed hippocampal samples were homogenized in lysis buffer containing a protease inhibitor cocktail (Applygen, Beijing, China). The levels of ACE, ACE2, Ang I, Ang II, and Ang-(1-7) were measured in the supernatants using a Parameter™ ACE (Nanjing Jiancheng Bioengineering Institute, China), ACE2 (Anaspec, USA), Ang I (R&D Systems, USA), Ang II (R&D Systems, USA), or Ang-(1-7) (R&D Systems, USA) immunoassay kit, respectively, according to the manufacturer's instructions. The absorbance at 405 nm was read using a SpectraMax M4 microplate reader (Molecular Devices, USA), and the concentration was determined by fitting curves using SoftMax Pro software (version 5.0, Molecular Devices, USA). Five rats from each group were randomly selected for ELISA.
Feng P., Wu Z., Liu H., Shen Y., Yao X., Li X, & Shen Z. (2020). Electroacupuncture Improved Chronic Cerebral Hypoperfusion-Induced Anxiety-Like Behavior and Memory Impairments in Spontaneously Hypertensive Rats by Downregulating the ACE/Ang II/AT1R Axis and Upregulating the ACE2/Ang-(1-7)/MasR Axis. Neural Plasticity, 2020, 9076042.
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7

Angiotensin II-Induced Cardiac Remodeling

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Wild-type mice and PAI-1−/− mice were infused with AngII (1 μg/kg/min; Sigma-Aldrich) or a vehicle control (PBS) using an osmotic minipump (Alzet, model 1002) as previously described18 (link); 9 to 10 male and female mice (16–20 weeks old) were used per group. Pumps remained implanted for 7 or 14 days. An additional group of 6 PAI-1−/− mice were infused with AngII as described earlier but also received injections of 300 μg/kg recombinant human BMP-7 in PBS (R&D Systems) every other day starting the day of minipump implantation. Arterial pressures were measured using the Coda2 noninvasive cuff system (Kent Scientific) for ≥8 consecutive measurements. Heart and tibia were extracted at day 7 or 28 after initiation of AngII. Transthoracic 2-dimensional M-mode echocardiography was performed using the Vevo770 (VisualSonics) equipped with a 30-MHz transducer. Mice were lightly anesthetized using a mixture of 1.5% isofluorane and oxygen (2 L/min) to ensure heart rates >400 to 450 bpm. Change in left ventricular area during the cardiac cycle was determined by endocardial border tracings to determine ejection fraction (average of parasternal long and short axis measurements). M-mode tracings were used to measure left ventricular systolic and diastolic chamber dimensions. The mean value from 3 beats was determined for each parameter.
Flevaris P., Khan S.S., Eren M., Schuldt A.J., Shah S.J., Lee D.C., Gupta S., Shapiro A.D., Burridge P.W., Ghosh A.K, & Vaughan D.E. (2017). Plasminogen Activator Inhibitor Type I Controls Cardiomyocyte Transforming Growth Factor-β and Cardiac Fibrosis. Circulation, 136(7), 664-679.
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8

Smooth Muscle Cells Response to Angiotensin II

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Human aortic smooth muscle cells (VSMCs) (ScienCell, no. 6110) were cultured in Smooth Muscle Cell Medium containing 2% fetal bovine serum (FBS), fibroblast growth factor-2, hydrocortisone, apo-transferrin, insulin, and insulin-like growth factor-1 (ScienCell, no. 1101). Ang II (100 nmol/L) was used to stimulate VSMCs for 1, 3 or 5 days. VSMCs were serum-starved for 24 h before stimulated by Ang II (R&D Systems, Minneapolis, MN, USA). Human embryonic kidney 293A cells were purchased from ATCC (Manassas, VA, USA) and were cultured in high glucose DMEM supplemented with 10% FBS.
Ma Y., Zheng B., Zhang X.H., Nie Z.Y., Yu J., Zhang H., Wang D.D., Shi B., Bai Y., Yang Z, & Wen J.K. (2021). circACTA2 mediates Ang II-induced VSMC senescence by modulation of the interaction of ILF3 with CDK4 mRNA. Aging (Albany NY), 13(8), 11610-11628.
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9

Molecular Mechanisms of Ang-II Signaling

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Ang-II and U0126 were purchased from R&D systems (Minneapolis, MN, USA). Sirt7, GAPDH, α-SMA, paxillin, phospho-Smad2, phospho-Smad3, Smad2/3, GFP and Vinculin antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). LaminB, PAI-1, FN, P-Thr/Ser and Collagen I antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). HRP-conjugated secondary antibodies were purchased from Santa Cruz Biotechnology.
Wang H., Liu S., Liu S., Wei W., Zhou X., Lin F., Wang J., Chen J., Zhang G, & Pang Y. (2017). Enhanced expression and phosphorylation of Sirt7 activates smad2 and ERK signaling and promotes the cardiac fibrosis differentiation upon angiotensin-II stimulation. PLoS ONE, 12(6), e0178530.
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10

Comprehensive Biomarker Evaluation in Kidney Injury

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The serum levels of BUN and creatinine were measured using automated, standardized procedures (Roche Hitachi 917/747, Mannheim, Germany). The levels of kidney injury molecule-1 (KIM-1; R&D Systems, Minneapolis, MN, USA), cytokine-induced neutrophil chemoattractant-1 (CINC-1; R&D Systems, Minneapolis, MN, USA), neutrophil gelatinase-associated lipocalin (NGAL, R&D Systems, Minneapolis, MN, USA), corticosterone (BioVendor, Germany), norepinephrine (LDN, Nordhorn, Germany), epinephrine (LDN, Nordhorn, Germany), and angiotensin II (Ang II, R&D Systems, Minneapolis, MN, USA) were determined through enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer’s instructions.
Chang C.Y., Pan P.H., Li J.R., Ou Y.C., Liao S.L., Chen W.Y., Kuan Y.H, & Chen C.J. (2021). Glycerol Improves Intracerebral Hemorrhagic Brain Injury and Associated Kidney Dysfunction in Rats. Antioxidants, 10(4), 623.
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