- PA5-27297 - Invitrogen Antibodies ANGPT2 antibody

Submitted references Direct Thrombin Inhibitor Dabigatran Compromises Pulmonary Endothelial Integrity in a Murine Model of Breast Cancer Metastasis to the Lungs; the Role of Platelets and Inflammation-Associated Haemostasis.

Smeda M, Stojak M, Przyborowski K, Sternak M, Suraj-Prazmowska J, Kus K, Derszniak K, Jasztal A, Kij A, Kurpinska A, Kieronska-Rudek A, Wojnar-Lason K, Buczek E, Mohaissen T, Chlopicki S
Frontiers in pharmacology 2022;13:834472

Tumor Angiogenesis Is Differentially Regulated by Phosphorylation of Endothelial Cell Focal Adhesion Kinase Tyrosines-397 and -861.

Pedrosa AR, Bodrug N, Gomez-Escudero J, Carter EP, Reynolds LE, Georgiou PN, Fernandez I, Lees DM, Kostourou V, Alexopoulou AN, Batista S, Tavora B, Serrels B, Parsons M, Iskratsch T, Hodivala-Dilke KM
Cancer research 2019 Sep 1;79(17):4371-4386

Dual antiplatelet therapy with clopidogrel and aspirin increases mortality in 4T1 metastatic breast cancer-bearing mice by inducing vascular mimicry in primary tumour.

Smeda M, Kieronska A, Proniewski B, Jasztal A, Selmi A, Wandzel K, Zakrzewska A, Wojcik T, Przyborowski K, Derszniak K, Stojak M, Kaczor D, Buczek E, Watala C, Wietrzyk J, Chlopicki S
Oncotarget 2018 Apr 3;9(25):17810-17824

Endothelial cell-derived angiopoietin-2 is a therapeutic target in treatment-naive and bevacizumab-resistant glioblastoma.

Scholz A, Harter PN, Cremer S, Yalcin BH, Gurnik S, Yamaji M, Di Tacchio M, Sommer K, Baumgarten P, Bähr O, Steinbach JP, Trojan J, Glas M, Herrlinger U, Krex D, Meinhardt M, Weyerbrock A, Timmer M, Goldbrunner R, Deckert M, Braun C, Schittenhelm J, Frueh JT, Ullrich E, Mittelbronn M, Plate KH, Reiss Y
EMBO molecular medicine 2016 Jan 1;8(1):39-57

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Angiopoietin 2 using 30 µg of HepG2 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with an Angiopoietin 2 polyclonal antibody (Product # PA5-27297) at a dilution of 1:5000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Angiopoietin 2 using 50 µg of mouse spleen lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with an Angiopoietin 2 polyclonal antibody (Product # PA5-27297) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Angiopoietin 2 in Mouse lung extracts (50 µg) and HUVEC cell extracts (30 µg). Samples were separated by 7.5% SDS-PAGE and the membrane was probed with Angiopoietin 2 Polyclonal antibody (Product # PA5-27297) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using Angiopoietin 2 Polyclonal Antibody (Product # PA5-27297). Whole cell extract (30 µg) was separated by 7.5% SDS-PAGE, and the membrane was blotted with Angiopoietin 2 Polyclonal Antibody (Product # PA5-27297) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of Angiopoietin-2 was achieved by transfecting HeLa with Angiopoietin-2 specific siRNAs (Silencer® select Product # s1360, s1359). Western blot analysis (Fig. a) was performed using Whole-cell extracts from the Angiopoietin-2 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Angiopoietin 2 Polyclonal Antibody (Product # PA5-27297, 1:1000) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). Densitometric analysis of this western blot is shown in the histogram (Fig. b). Decrease in signal upon siRNA mediated knockdown confirms that antibody is specific to Angiopoietin-2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-Angiopoietin 2 Polyclonal Antibody (Product # PA5-27297) and a 65 kDa band corresponding to Angiopoietin-2 was observed across the cells and tissues tested. Whole cell extracts (30 µg lysate) of HUVEC (Lane 1), HAEC (Lane 2), HeLa (Lane 3), Mouse Aorta (Lane 4), Rat Aorta (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Angiopoietin 2 (Product # PA5-27297) antibody at 1:250 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure EV1 Application of tissue microarrays ( TMA s) for the analysis of human glioma samples A, B Biopsies of 303 human gliomas ( WHO grade II , n = 16, WHO grade III , n = 35, and WHO grade IV , n = 252) were spotted on microscope slides (A) and processed for automated anti-Ang-2 immunohistochemistry (Ventana Benchmark platform) (B). Scale bar (B): 200 mum, inset: 50 mum. C Ang-2 expression was assessed by applying a semiquantitative scoring system (Harter et al , 2010 ). D Spatial expression of Ang-2 in brain specimens from glioblastoma patients was scored in normal-appearing gray matter ( NAGM ) ( n = 48), normal-appearing white matter ( NAWM ) ( n = 18), infiltration zone ( n = 39), and tumor center ( n = 62). Data information: In (C, D), for statistical analysis, Kruskal-Wallis test (followed by Dunn's post-test) was applied. *** P < 0.005. Whisker Box plots displaying median, 25-75 th percentile, upper and lower quartile.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIGURE 3 Effects of dabigatran on lung permeability and inflammation markers in the lungs of BALB/c mice injected with 4T1 breast cancer cells. Mice were treated with vehicle (black symbols) or dabigatran etexilate (grey symbols) as described in Section 2.5 . (A) depicts retention of Evans blue (EB) in the lung parenchyma, reflecting lung permeability. Control mice did not receive cancer cells and were euthanized at the end of the dabigatran pre-treatment to exclude the possibility that dabigatran affected pulmonary permeability of healthy animals before their inoculation with 4T1 cancer cells. After dabigatran/vehicle pre-treatment, the other mice were injected intravenously with 4T1 breast cancer cells or the vehicle and euthanized 24 h, 2 days, and 7 days after injection. (B-D) show the levels of Ang-2, E-selectin, and MMP-9, respectively. (E) shows representative Western blot images of Ang-2, E-selectin, and MMP-9 with densitometric data after their normalization to the total protein used as loading control based on the stain-free technique as described in Materials and Methods . For (A-D) , statistical analysis was performed with two-way ANOVA and an appropriate post hoc test. The symbols *, **, and *** denote statistical significance at p < .05, p < .01, and p < .001, respectively, and the data are depicted as the median +- IQR.

PA5-27297

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