goat anti mouse 412 lyve1 antibody  (R&D Systems)

Journal: Endocrinology

Article Title: GH Alters Lymphatic Vessels in Female Mice and STAT5 Phosphorylation in Human Lymphatic Endothelial Cells

doi: 10.1210/endocr/bqaf194

Figure Lengend Snippet: LVD quantification using IHC on the hearts of transgenic mice with perturbed GH signaling. (A) Hematoxylin and eosin staining of mouse heart sections from each genotype. (B) Representative images from IHC staining using 2 primary antibodies against LYVE1 and CD31. Fluorescent detection using a red Cy5 or green Alexa488 fluorophore bound to a secondary antibody was used to visualize LYVE1 and CD31, respectively. (C) Fluorescent IHC combined with brightfield imaging using the previously mentioned antibodies toward specific protein targets (LYVE1 and CD31) and using isotype control antibodies. (D, E) The LYVE1-positive area and total area were measured using a pixel classifier in QuPath across 3 to 5 sections per mouse from 3 mice per group (n = 3). The mean LVD ± SEM is shown in each bar graph. Group comparisons were performed using an unpaired t -test, with P -values displayed above the graph. Abbreviations: IHC, immunohistochemistry; LVD, lymphatic vascular density; LYVE1, lymphatic vessel-hyaluronic acid receptor 1.

Article Snippet: The primary antibodies were prepared in TBS-T with 1% BSA at the following concentrations: 4 ug/mL of goat anti-mouse lymphatic vessel-hyaluronic acid receptor 1 (LYVE1; R&D Systems Cat# AF2125, RRID: AB_2297188 ), 3 ug/mL of rabbit anti-mouse CD31 (Proteintech Cat# 28083-1-AP, RRID: AB_2881055 ), or 4 ug/mL goat isotype control antibody (R&D Systems Cat# AB-108-C, RRID: AB_354267 ) or 3 ug/mL rabbit isotype control antibodies (R&D Systems Cat# AB-105-C, RRID: AB_354266 ).

Techniques: Transgenic Assay, Staining, Immunohistochemistry, Imaging, Control

Journal: Endocrinology

Article Title: GH Alters Lymphatic Vessels in Female Mice and STAT5 Phosphorylation in Human Lymphatic Endothelial Cells

doi: 10.1210/endocr/bqaf194

Figure Lengend Snippet: Skin histology of the uncut and cut sides 1 month after surgery in transgenic mice with altered GH signaling. Representative H&E and fluorescent IHC images of cut (A) and uncut (B) skin from the bGH, WT littermate from the GHA cohort, and GHA mice. The H&E image is of the full skin thickness at 10×, while the IHC image is of the dermis at 20×. The IHC staining used 1 primary antibody against LYVE1 and 1 secondary conjugated to the Cy5 fluorophore along with DAPI staining of the nuclei. The LYVE1-positive area and total area were measured using a pixel classifier in QuPath across 3 to 5 sections per mouse from 2 to 3 mice per group (n = 2-3). To assess statistical differences between the cut and uncut sides and between WT and transgenic animals, a mixed-effects 2-way ANOVA was used. No statistical difference in LVD was observed between WT and transgenic animals in the dermis or hypodermis or between the cut and uncut sides in the bGH (C) and GHA (E) groups; therefore, P -values from post hoc analysis were not calculated or displayed. Bar graphs represent mean ± SEM. (D) Brightfield imaging was combined with fluorescent imaging of LYVE1 staining or isotype control antibody staining to assess for antibody specificity. Abbreviations: bGH, bovine GH; DAPI, 4',6-diamidino-2-phenylindole; GHA, GH antagonist; H&E, hematoxylin and eosin; IHC, immunohistochemistry; LVD, lymphatic vascular density; LYVE1, lymphatic vessel-hyaluronic acid receptor 1; WT, wild-type.

Article Snippet: The primary antibodies were prepared in TBS-T with 1% BSA at the following concentrations: 4 ug/mL of goat anti-mouse lymphatic vessel-hyaluronic acid receptor 1 (LYVE1; R&D Systems Cat# AF2125, RRID: AB_2297188 ), 3 ug/mL of rabbit anti-mouse CD31 (Proteintech Cat# 28083-1-AP, RRID: AB_2881055 ), or 4 ug/mL goat isotype control antibody (R&D Systems Cat# AB-108-C, RRID: AB_354267 ) or 3 ug/mL rabbit isotype control antibodies (R&D Systems Cat# AB-105-C, RRID: AB_354266 ).

Techniques: Transgenic Assay, Immunohistochemistry, Staining, Imaging, Control