protein based elisa like assay  (R&D Systems)

Journal: Scientific Reports

Article Title: Extracellular Superoxide Dismutase Regulates Early Vascular Hyaluronan Remodeling in Hypoxic Pulmonary Hypertension

doi: 10.1038/s41598-019-57147-7

Figure Lengend Snippet: Changes in hyaluronan content during hypoxia. Changes in HA levels during hypoxia, determined by HABP-based ELISA-like assay in ( A ) lung tissue after alveolar lavage in WT (gray bars) and SOD3KO (KO, red bars) mice; ( B ) serum of WT and SOD3KO mice; ( C ) lung tissue of control SMC-SOD3Flox (Lox, gray bars) and SMC-SOD3cKO (cKO, red bars) mice after induction with tamoxifen; ( D ) serum of SMC-SOD3Flox and SMC-SOD3cKO mice. All mice were 8–10 weeks old. Males and females were equally represented in the WT and SOD3KO groups; all SMC-SOD3Flox and SMC-SOD3cKO mice were males. Data represented as mean ± SEM *p < 0.05 **p < 0.01 by 2-way ANOVA.

Article Snippet: HA concentrations were assayed using the Aggrecan HA-binding protein (HABP) G1 Link-domain based ELISA-like assay (Quantikine, R&D Systems; Minneapolis, MN).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Scientific Reports

Article Title: Extracellular Superoxide Dismutase Regulates Early Vascular Hyaluronan Remodeling in Hypoxic Pulmonary Hypertension

doi: 10.1038/s41598-019-57147-7

Figure Lengend Snippet: Impact of hypoxia and SOD3 on perivascular hyaluronan and arterial muscularization. Relative changes in HA in the vascular adventitial vs . parenchymal compartments of the lung between normoxia and 14 days of hypoxia. Quantitation of HA in HABP-stained lung sections was achieved by DAB/methyl green color vector deconvolution in ImageJ followed by signal-to-perimeter measurement of vessel edges (5–10 vessels per HPF; 5 HPF per lung) normalized to alveolar background generated by cropping away all bronchovascular structures. ( A ) Representative images of vascular HA staining after image processing. ( B ) Mean alveolar background HA staining intensity. ( C ) HA intensity expressed as vascular HA (<50 or 50–100 micron diameter) relative to the alveolar HA content in normoxic and hypoxic WT and SOD3KO (KO) mice. ( D ) Representative histo-immunologic co-staining of HABP and αSMA in lung slices. Vessels are demarcated by black arrows. ( E ) Correlation between HA-coated and de novo muscularized small vessels (≤50 microns – left panel; 50–100 microns – right panel) during hypoxic PH. Color vectors were deconvolved in ImageJ (DAB/methyl green) and independently counted in a blinded fashion. Legend: WT normoxic (open black circles), 14d hypoxic (light gray closed circles), and 35d hypoxic (darker grey closed circles); SOD3KO (open red circles), 14d hypoxic (light pink closed circles) and 35d (darker pink closed circles). 5–10 randomly selected HPF were analyzed per animal. Data represented as mean ± SEM *p < 0.05 **p < 0.01 by 2-way ANOVA.

Article Snippet: HA concentrations were assayed using the Aggrecan HA-binding protein (HABP) G1 Link-domain based ELISA-like assay (Quantikine, R&D Systems; Minneapolis, MN).

Techniques: Quantitation Assay, Staining, Plasmid Preparation, Generated