mouse anti vdbp  (Santa Cruz Biotechnology)

Journal: Frontiers in Physiology

Article Title: A new SO 2 probe ZSO targeting VDBP inhibits high glucose induced endothelial cell senescence and calcification

doi: 10.3389/fphys.2025.1719853

Figure Lengend Snippet: ZSO inhibits high glucose-induced vascular endothelial cell senescence and calcification through downregulating VDBP protein levels. (A) Endothelial cells were treated with high glucose (30 mM) and various concentrations of ZSO (1, 5, 10 μM) for 24 h. RT-qPCR was then performed to quantify VDBP mRNA level, and the results were statistically analyzed. (B) Endothelial cells were treated with DOM and various concentrations of ZSO (1, 5, 10 μM) for 4 days. RT-qPCR was then performed to quantify VDBP mRNA level, and the results were statistically analyzed. (C) Endothelial cells were treated with high glucose (30 mM) and ZSO (5 μM) for 24 h, followed by Western blot analysis of VDBP protein level. (D) Statistical analysis of VDBP protein level under senescence conditions. (E) Endothelial cells were treated with 5 μM ZSO under calcification-inducing conditions for 4 days, followed by Western blot analysis of VDBP protein level. (F) Quantitative analysis of VDBP protein level under calcification conditions. (G) Immunohistochemical staining of VDBP in mouse thoracic aorta sections, with brown coloration indicating positive immunoreactivity. (H) Quantitative analysis of VDBP-positive staining intensity in vascular tissues, with corresponding statistical graphs. (I) Measurement of VDBP protein level in mouse serum using an ELISA kit, followed by statistical analysis. (J) Endothelial cells were treated with high glucose (30 mM) and 5 μM ZSO for 24 h, and the ubiquitination level of VDBP was assessed. (K) Quantification of VDBP ubiquitination level. ns p > 0.05, *p < 0.05, **p < 0.01, n = 3.

Article Snippet: Beads were washed, and Western blot was used for detection, first incubated with ubiquitination antibody (Abcam, ab137031), developed, and then incubated with VDBP antibody (Santa Cruz, sc-365441) and developed.

Techniques: Quantitative RT-PCR, Western Blot, Immunohistochemical staining, Staining, Enzyme-linked Immunosorbent Assay, Ubiquitin Proteomics

Journal: Journal of Inflammation Research

Article Title: Vitamin D Binding Protein: A Potential Factor in Geriatric COVID-19 Acute Lung Injury

doi: 10.2147/JIR.S470097

Figure Lengend Snippet: The VDBP expression in injured alveolar epithelial cells. ( A ) The VDBP protein level in LPS-induced injury of alveolar epithelial cells at different time points. The representative Western blot results and the relative expression of VDBP from the Western blot were shown. ( B ) The VDBP gene expression in the LPS-induced injury of alveolar epithelial cells. The q-PCR was used to analyze the relative expression of genes. Columns and error bars represented mean ± SEM. ** P < 0.01, *** P < 0.001. VDBP: Vitamin D-binding protein, LPS: Lipopolysaccharide, NC: negative control, GAPDH: glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Then separate equal amounts of proteins on 12% SDS-PAGE for Western blot using antibodies against VDBP (1:2000 dilution, Proteintech, US) and GAPDH (1:5000 dilution, Proteintech, US), the protein was observed using the ECL chemiluminescence method.

Techniques: Expressing, Western Blot, Gene Expression, Binding Assay, Negative Control