goat anti mouse tie2 r d systems af762  (R&D Systems)

Journal: Development (Cambridge, England)

Article Title: TIE1 dependent lymphatic vascular remodeling is mediated by its second tyrosine kinase domain.

doi: 10.1242/dev.204469

Figure Lengend Snippet: Fig. 1. TIE2 and TIE1 expression in LECs of developing and mature lymphatics.

Article Snippet: Primary antibodies for immunohistochemistry Antibody Supplier Catalogue # Application Goat anti-mouse VEGFR3 R&D Systems AF743 1:50 Goat anti-human TIE1 R&D Systems AF619 1:100 Goat anti-mouse TIE2 R&D Systems AF762 1:200 Goat anti-mouse integrinα9 R&D Systems AF3827 1:50 Goat anti-VE-Cadherin R&D Systems AF1002 1:400 Goat anti-human Prox1 R&D Systems AF2727 1:100 Sheep anti-mouse FOXC2 R&D Systems AF6989 1:200 Rabbit anti-GFP Invitrogen A11122 1:200 Rabbit anti-FOXO1 Cell Signaling 2880 1:200 Rabbit anti-Lyve1 abcam Ab14917 1:800 Rabbit anti-Prox1 abcam Ab37128 1:250 Rat anti-mouse CD31 PD Pharmingen 557355 1:200 Rat anti-endomucin abcam ab106100 1:200 Development: doi:10.1242/dev.204469: Supplementary information D ev el o pm en t • S up pl em en ta ry in fo rm at io n

Techniques: Expressing

Journal: Development (Cambridge, England)

Article Title: TIE1 dependent lymphatic vascular remodeling is mediated by its second tyrosine kinase domain.

doi: 10.1242/dev.204469

Figure Lengend Snippet: Fig. 2. LEC specific deletion of Tie1 but not Tie2 results in defective lymphatic

Article Snippet: Primary antibodies for immunohistochemistry Antibody Supplier Catalogue # Application Goat anti-mouse VEGFR3 R&D Systems AF743 1:50 Goat anti-human TIE1 R&D Systems AF619 1:100 Goat anti-mouse TIE2 R&D Systems AF762 1:200 Goat anti-mouse integrinα9 R&D Systems AF3827 1:50 Goat anti-VE-Cadherin R&D Systems AF1002 1:400 Goat anti-human Prox1 R&D Systems AF2727 1:100 Sheep anti-mouse FOXC2 R&D Systems AF6989 1:200 Rabbit anti-GFP Invitrogen A11122 1:200 Rabbit anti-FOXO1 Cell Signaling 2880 1:200 Rabbit anti-Lyve1 abcam Ab14917 1:800 Rabbit anti-Prox1 abcam Ab37128 1:250 Rat anti-mouse CD31 PD Pharmingen 557355 1:200 Rat anti-endomucin abcam ab106100 1:200 Development: doi:10.1242/dev.204469: Supplementary information D ev el o pm en t • S up pl em en ta ry in fo rm at io n

Techniques:

Journal: Developmental biology

Article Title: Loss of Flow Responsive Tie1 Results in Impaired Aortic Valve Remodeling

doi: 10.1016/j.ydbio.2019.07.011

Figure Lengend Snippet: Key Resource Table (Antibodies for IHC)

Article Snippet: Ultrastructural analysis of endothelial morphology was examined with transmission electron microscopy (EM) as described ( Odelin et al., 2014 ). table ft1 table-wrap mode="anchored" t5 caption a7 Antibodies SOURCE IDENTIFIER Working concentration Rabbit anti-pHH3 Upstate Cat#06–570; PRID:AB_310177 1:100 Rat anti-BrdU Abcam Cat#ab6326; PRID:AB_305426 1:100 Rabbit anti Cleaved Caspase-3 (Asp175) Cell Signaling Cat#9664; PRID:AB_2070042 1:400 Goat anti Tie1 R&D Systems Cat#AF619; PRID:AB_355481 1:50 Goat anti-mouse Tie2 R&D Systems Cat#AF762; PRID:AB_2203220 1:400 Rat anti-mouse CD31 Pharmingen, Cat#557355; PRID:AB_396660 1:200 Mouse anti Collagen III Abcam Cat#ab6310; PRID:AB_305413 1:400 Rabbit anti Collagen I Abcam Cat#ab34710; PRID:AB_731684 1:100 Rabbit anti-Erg1 Abcam Cat#ab92513; PRID:AB_2630401 1:100 Goat anti-VE-Cadherin R&D Systems Cat#AF1002; PRID:AB_2077789 1:400 Rabbit anti-Versican (GAGβ) Chemicon Cat#AB1033; PRID:AB_90462 1:1500 Hyaluronic acid binding protein, biotinylated (HABP) Millipore Cat#385911 1:100 Goat anti-Sox9 Cell Signaling Cat#82630 1:100 Rabbit anti Versican V0,V1 Neo (DPEAAE) ThermoFisher Cat#PA1-1748A; PRID:AB_2304324 1:1500 Sheep anti-Foxc2 R&D Systems Cat#AF6989; PRID:AB_10973139 1:200 NL557 Conjugated donkey anti-sheep IgG R&D Systems Cat#NL010; PRID:AB_884220 1:600 Open in a separate window Key Resource Table (Antibodies for IHC) 2.4.

Techniques: Concentration Assay, Binding Assay

Journal: Developmental biology

Article Title: Loss of Flow Responsive Tie1 Results in Impaired Aortic Valve Remodeling

doi: 10.1016/j.ydbio.2019.07.011

Figure Lengend Snippet: Polarized expression of Tie1 and other flow sensitive endothelial markers in the remodeling aortic valves. A, Tie1 (red), Cdh5 (red), CD31 (green) and Tie2 (red) expression in remodeling aortic valves was detected by immunostaining Tie1fl/fl hearts at E14.5, E16.5, and E18.5. B and C, AoV sections of P0 Tie1fl/fl heart were co-immunostained for Tie1 (red) and CD31 (green) or Cdh5 (red) and CD31 (green). D, P0 Tie1+/lacz heart AoV sections were stained for (β-Gal, β-galactosidase) with X-gal (blue) and then with Tie2 antibody (red, E). DAPI (blue) denotes nuclei. Scale bars: 50 μm. From E16.5, Tie1 and Cdh5 begin to have higher expression on the arterial sides (arrows); in contrast, CD31 and Tie2 have higher expression on the ventricular sides (arrowheads). F, Quantification of relative expression of Tie1, Cdh5, CD31 and Tie2 in remodeling aortic valves: arterial side (predicted to face oscillatory shear stress) vs. ventricular side (predicted to face high laminar shear stress). *P<0.05, **P<0.0,1***P<0.001 (all compared with E14.5).

Article Snippet: Ultrastructural analysis of endothelial morphology was examined with transmission electron microscopy (EM) as described ( Odelin et al., 2014 ). table ft1 table-wrap mode="anchored" t5 caption a7 Antibodies SOURCE IDENTIFIER Working concentration Rabbit anti-pHH3 Upstate Cat#06–570; PRID:AB_310177 1:100 Rat anti-BrdU Abcam Cat#ab6326; PRID:AB_305426 1:100 Rabbit anti Cleaved Caspase-3 (Asp175) Cell Signaling Cat#9664; PRID:AB_2070042 1:400 Goat anti Tie1 R&D Systems Cat#AF619; PRID:AB_355481 1:50 Goat anti-mouse Tie2 R&D Systems Cat#AF762; PRID:AB_2203220 1:400 Rat anti-mouse CD31 Pharmingen, Cat#557355; PRID:AB_396660 1:200 Mouse anti Collagen III Abcam Cat#ab6310; PRID:AB_305413 1:400 Rabbit anti Collagen I Abcam Cat#ab34710; PRID:AB_731684 1:100 Rabbit anti-Erg1 Abcam Cat#ab92513; PRID:AB_2630401 1:100 Goat anti-VE-Cadherin R&D Systems Cat#AF1002; PRID:AB_2077789 1:400 Rabbit anti-Versican (GAGβ) Chemicon Cat#AB1033; PRID:AB_90462 1:1500 Hyaluronic acid binding protein, biotinylated (HABP) Millipore Cat#385911 1:100 Goat anti-Sox9 Cell Signaling Cat#82630 1:100 Rabbit anti Versican V0,V1 Neo (DPEAAE) ThermoFisher Cat#PA1-1748A; PRID:AB_2304324 1:1500 Sheep anti-Foxc2 R&D Systems Cat#AF6989; PRID:AB_10973139 1:200 NL557 Conjugated donkey anti-sheep IgG R&D Systems Cat#NL010; PRID:AB_884220 1:600 Open in a separate window Key Resource Table (Antibodies for IHC) 2.4.

Techniques: Expressing, Immunostaining, Staining, Shear

Journal: Developmental biology

Article Title: Loss of Flow Responsive Tie1 Results in Impaired Aortic Valve Remodeling

doi: 10.1016/j.ydbio.2019.07.011

Figure Lengend Snippet: Nfatc1enCre mediates loxP excision exclusively in the valvular endocardium. A-D, X-gal staining of R26fslz;Nfatc1enCre heart sections shows that Nfatc1enCre-mediated recombination is restricted to the endothelium of the developing aortic valves (arrows) but not the endothelium of the artery (solid arrowheads), mesenchyme derived from EMT or ventricular endocardium (open arrowheads) at E12.5 (A), at E14.5 (B) and at P0 (C), and lacZ expression remains in the majority of the valvular endothelial cells throughout adulthood (D). E, Representative images of immuno-staining of Tie2-cko AoVs at E14.5 with anti-Tie1 antibody showing efficient specific Tie1 deletion in valvular endothelium (arrows) without change of normal expression of Tie1 in arterial endothelium (solid arrowheads) or ventricular endocardium (open arrowheads). F, Nfatc1enCre-mediated Tie1 deletion was not very efficient in a few (2 out of 9) mutant embryos. G and H, the same sections in E and F were co-stained for CD31, showing the integrity of endothelial cells. Yellow arrows indicate the mosaic Tie1 expression in the mutant valvular endothelium. DAPI, blue. Scale bars: 50μm in A-C, H-I and 200μm in D. I, The efficiency of Cre mediated Tie1 deletion in AoVs was assessed with qRT-PCR on valve leaflets of Tie1fl/fl (control) and Tie1fl/lz;Nfatc1enCre (Tie1-cko) mice at E14.5 and at P0. For normalization, Tie1 mRNA level in control mice at E14.5 was set as 1. ***P<0.001.

Article Snippet: Ultrastructural analysis of endothelial morphology was examined with transmission electron microscopy (EM) as described ( Odelin et al., 2014 ). table ft1 table-wrap mode="anchored" t5 caption a7 Antibodies SOURCE IDENTIFIER Working concentration Rabbit anti-pHH3 Upstate Cat#06–570; PRID:AB_310177 1:100 Rat anti-BrdU Abcam Cat#ab6326; PRID:AB_305426 1:100 Rabbit anti Cleaved Caspase-3 (Asp175) Cell Signaling Cat#9664; PRID:AB_2070042 1:400 Goat anti Tie1 R&D Systems Cat#AF619; PRID:AB_355481 1:50 Goat anti-mouse Tie2 R&D Systems Cat#AF762; PRID:AB_2203220 1:400 Rat anti-mouse CD31 Pharmingen, Cat#557355; PRID:AB_396660 1:200 Mouse anti Collagen III Abcam Cat#ab6310; PRID:AB_305413 1:400 Rabbit anti Collagen I Abcam Cat#ab34710; PRID:AB_731684 1:100 Rabbit anti-Erg1 Abcam Cat#ab92513; PRID:AB_2630401 1:100 Goat anti-VE-Cadherin R&D Systems Cat#AF1002; PRID:AB_2077789 1:400 Rabbit anti-Versican (GAGβ) Chemicon Cat#AB1033; PRID:AB_90462 1:1500 Hyaluronic acid binding protein, biotinylated (HABP) Millipore Cat#385911 1:100 Goat anti-Sox9 Cell Signaling Cat#82630 1:100 Rabbit anti Versican V0,V1 Neo (DPEAAE) ThermoFisher Cat#PA1-1748A; PRID:AB_2304324 1:1500 Sheep anti-Foxc2 R&D Systems Cat#AF6989; PRID:AB_10973139 1:200 NL557 Conjugated donkey anti-sheep IgG R&D Systems Cat#NL010; PRID:AB_884220 1:600 Open in a separate window Key Resource Table (Antibodies for IHC) 2.4.

Techniques: Staining, Derivative Assay, Expressing, Immunostaining, Mutagenesis, Quantitative RT-PCR, Control