human rage/ager antibody  (Bio-Techne corporation)

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Self-assembled elastin-like polypeptide fusion protein coacervates as competitive inhibitors of advanced glycation end-products enhance diabetic wound healing

doi: 10.1016/j.jconrel.2021.03.032

Figure Lengend Snippet: Effect of AGEs and RAGEs inhibitors on HUVEC number and ROS generation. (A) Viable cell number. Viable cell number was measured after 24 h of incubation in each condition and normalized to the BSA control = 100%. AGEs alone significantly decreased cell number (p = 0.0147). vRAGE-ELP, but not empty ELP, restored cell viability in a dose-dependent manner. NAC, an antioxidant, also recovered cell viability. N = 6. Statistics: One-way ANOVA followed by Tukey’s test: #p < 0.05, ### p < 0.001, * p < 0.05, ** p < 0.01, *** p < 0.001. Asterisks indicate comparisons with AGE group. (B) ROS levels. AGE stimulation increased ROS generation in HUVECs. The ROS level significantly decreased when cells were treated vRAGE-ELP or NAC, while empty ELPs had no effect. AGE = 100 μg/mL, vRAGE-ELP = 1 mg/mL, ELP = 1 mg/mL, NAC = 30 mM. Blue: Nuclei, Green: ROS. Scale bar = 200 μm.

Article Snippet: When cells became confluent, cells were treated with BSA (100 μg/mL), AGE (100 μg/mL) only, AGE (100 μg/mL) + vRAGE-ELP (1 mg/mL) (pre-incubated for 30 min), AGE (100 μg/mL) + ELP (1 mg/mL) (pre-incubated for 30 min), AGE (100 μg/mL) + NAC (30 mM) or AGE (100 μg/mL) + Human RAGE antibody (10 μg/mL, R&D Systems, MAB11451) and incubated at 37 °C for 6 h. Cells were then washed with fresh media twice and fixed in 2% paraformaldehyde in PBS for 20 min at room temperature.

Techniques: Incubation, Control

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Self-assembled elastin-like polypeptide fusion protein coacervates as competitive inhibitors of advanced glycation end-products enhance diabetic wound healing

doi: 10.1016/j.jconrel.2021.03.032

Figure Lengend Snippet: Effect of AGEs and RAGEs inhibitors on surface ICAM-1 expression. (A) Images of immunostaining. vRAGE-ELP coacervates suppressed ICAM-1 expression in AGE-stimulated cells. Blue: DAPI-stained nuclei. Red: antibody-stained ICAM-1. Scale bar = 20 μm. (B) Cell-based ELISA. vRAGE-ELP significantly reduced the expression of ICAM-1 (p < 0.0001). N = 6. Statistics: One-way ANOVA, Tukey’s test. #### p < 0.0001, **** p < 0.0001. Asterisks indicate comparisons with AGE group. BSA = 100 μg/mL. AGE = 100 μg/mL. vRAGE-ELP = 1 mg/mL. Empty ELP = 1 mg/mL. NAC = 30 mM. Anti-RAGE antibody = 10 μg/mL.

Article Snippet: When cells became confluent, cells were treated with BSA (100 μg/mL), AGE (100 μg/mL) only, AGE (100 μg/mL) + vRAGE-ELP (1 mg/mL) (pre-incubated for 30 min), AGE (100 μg/mL) + ELP (1 mg/mL) (pre-incubated for 30 min), AGE (100 μg/mL) + NAC (30 mM) or AGE (100 μg/mL) + Human RAGE antibody (10 μg/mL, R&D Systems, MAB11451) and incubated at 37 °C for 6 h. Cells were then washed with fresh media twice and fixed in 2% paraformaldehyde in PBS for 20 min at room temperature.

Techniques: Expressing, Immunostaining, Staining, In-Cell ELISA

Journal: ACS Omega

Article Title: Crosstalk between Aldosterone and Glycation through Rac-1 Induces Diabetic Nephropathy

doi: 10.1021/acsomega.3c05085

Figure Lengend Snippet: G.HSA + Aldo effect on RAGE and NF-κB protein expressions in renal cells. HEK293T cells, after 24 h treatment of glycated samples (200 μg/mL), were lysed and lysates were used for Western blotting - (a) RAGE, NF-κB and β-actin expressions in cells treated with glycated HSA samples; ratio of (b) RAGE and (c) NF-κB to β-actin by densitometry analysis. The data are presented as mean ± SD, n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, *indicates p value in comparison to HSA; # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001, # indicates the p-value in comparison to G.HSA.

Article Snippet: Primary antibodies for CML (MAB3247), RAGE (MAB11451), NF-κB (SC8008), β-actin (SC130301), and secondary antibody HRP conjugated goat antimouse IgG were procured from R&D systems (Minneapolis, United States) and Santacruz (Dallas, United States).

Techniques: Western Blot, Comparison