mouse vegf quantikine colorimetric sandwich elisa kit  (R&D Systems)

Journal: Biology Open

Article Title: The autotaxin-LPA axis promotes membrane trafficking and secretion in yolk sac visceral endoderm cells

doi: 10.1242/bio.060081

Figure Lengend Snippet: Enpp2 knockout impairs secretion of angiogenic factors from the yolk sac. (A-B) Concentration of TGF-β1 (A) and VEGF (B) in the ECF of Enpp2 +/− and Enpp2 −/− embryos at E8.5 ( n =4 in [A], n =3 in [B], unpaired t -test). (C-D) Amount of TGF-β1 and VEGF secreted from the yolk sac. After the yolk sac was incubated for 16 h at 37°C in a culture medium, TGF-β1 and VEGF concentrations in the medium were measured by means of ELISA. Secretion of TGF-β1 and VEGF was reduced in the Enpp2 −/− yolk sacs ( n =3, unpaired t -test). (E) RT-PCR of Vegfa and Tgfb1 in the embryo and yolk sac of Enpp2 +/+ , Enpp2 +/− , and Enpp2 −/− at E8.5. RT- indicates a negative control in which a reverse-transcriptase was omitted from the reaction. (F) Western blot analysis of VEGF in the embryo and yolk sac of E8.5 mice. Abbreviation: n.s., not significant.

Article Snippet: Amounts of VEGF-A and TGF-β1 in the ECF and yolk sac were determined by use of a mouse VEGF Quantikine Colorimetric Sandwich ELISA kit (R&D Systems) and a TGF-β1 E MAX ImmunoAssay System (Promega), respectively.

Techniques: Knock-Out, Concentration Assay, Incubation, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Negative Control, Reverse Transcription, Western Blot

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: VEGFR-1/Flt-1 inhibition increases angiogenesis and improves muscle function in a mouse model of Duchenne muscular dystrophy

doi: 10.1016/j.omtm.2021.03.013

Figure Lengend Snippet: Characterization of 21B3 (A and B) Representative curves from the Biacore affinity analysis of 21B3 binding to (A) mouse or (B) human Flt-1. The observed response for the kinetic titration series is shown in red, and the fit to a 1:1 interaction model is shown in black. (C and D) Competition ELISA of VEGF binding to (C) mouse (n = 2 representative images) or (D) human sFlt-1 (n = 3) in the presence of increasing concentrations of 21B3. Data are presented as mean and standard deviation. (E) Inhibition of VEGFR-2 phosphorylation by mouse and human sFlt-1 in HUVECs. The VEGF bar represents 100% of the phospho-VEGFR2 signal. The data thus represent a percent of the signal seen with VEGF alone; for example, the addition of sFlt-1 inhibited the signal to ∼25% compared with VEGF alone. Phosphorylation increased with the addition of 10-fold molar excess mouse 21B3 and 4-fold excess human 21B3 compared with sFlt-1 (n = 3–6; data represent mean and standard deviation). ∗∗∗∗p < 0.0001 by ordinary ANOVA with multiple comparisons. (F) Ribbon model of the sFlt-1:VEGF complex (PDB 5T89 ) highlighting the 21B3 epitope. sFlt-1 is shown in blue and cyan for each member of the complex dimer, and the VEGF dimer is shown as dark and light green. The two peptides in sFlt-1 domain 2 that were shown to interact with 21B3 using hydrogen-deuterium exchange are depicted in magenta. RU, response unit.

Article Snippet: Free VEGF in diluted serum samples was quantified in a solid-phase sandwich ELISA, as per manufacturer protocol (MMV00; R&D Systems).

Techniques: Binding Assay, Titration, Enzyme-linked Immunosorbent Assay, Standard Deviation, Inhibition, Phospho-proteomics