hfn7 1 monoclonal antibody  (Novus Biologicals)

Journal: The American journal of pathology

Article Title: Overexpression of Corin Ameliorates Kidney Fibrosis through Inhibition of Wnt/β-Catenin Signaling in Mice.

doi: 10.1016/j.ajpath.2023.09.008

Figure Lengend Snippet: Figure 1 Wnt/b-catenin signaling is activated and corin expression is decreased along with the increased expression of b-catenin in mouse kidney after unilateral ureteral obstruction (UUO). A and B: Western blot analyses of kidney expression of several proteins, including Wnt1, a-smooth muscle actin (a- SMA), type I collagen, fibronectin, Snail, and plasminogen activator inhibitor-1 (PAI-1), in sham and obstructed kidneys at day 7 after UUO. Representative Western blot analysis (A) and quantitative data relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH; B) are presented. C: Representative mi- crographs using immunohistochemical staining for corin and active b-catenin in kidney tissues at days 4 and 7 after UUO. Boxed areas show magnification of images. Arrows indicate corin- or active b-cateninepositive tubular epithelial cells. Arrowheads in the enlarged boxed areas indicate positive staining. D and E: Western blot analysis of kidney expression of corin and active b-catenin protein at days 4 and 7 after UUO. Representative Western blot analysis (D) and quantitative data (E) are presented. F: Linear regression graph of kidney corin and b-catenin protein expression at days 0, 4, and 7 after UUO. The Spearman correlation coefficient (R2) is shown. G: Representative micrographs using immunohistochemical staining for proprotein convertase subtilisin/kexin-6 (PCSK6) in kidney tissues at days 4 and 7 after UUO. Boxed area shows magnification of image. Arrows indicate PCSK6-positive tubular epithelial cells. Arrowheads in the enlarged boxed area indicate positive staining. H and I: Western blot analysis of PCSK6 protein in kidney at days 4 and 7 after UUO. Representative Western blot analysis (H) and quantitative data (I) are presented. Data are the means SEM (B, E, and I). n Z 6 animals per group (B, E, and I). *P < 0.05, **P < 0.01, and ***P < 0.001. Scale bars: 50 mm (C and G, main images); 200 mm (C and G, enlarged images).

Article Snippet: HKC-8 cells were treated with recombinant human transforming growth factor (TGF)-b1 ajp.amjpathol.org - The American Journal of Pathology Table 1 Used Antibodies in Experiments Antibodies Dilution for IHC/IF Dilution for WB Company Product code Anti-Wnt1 monoclonal antibody e 1:1000 Abcam (Cambridge, UK) ab15251 Anti-corin monoclonal antibody IHC: 1:200 1:1000 Abcam ab274323 Antieb-catenin nonphosphorylated (active) S37/T41 monoclonal antibody IHC: 1:2000 1:1000 Abcam ab246504 GAPDH monoclonal antibody e 1:5000 ProteinTech (Chicago, IL) 60004-1-Ig HA tag polyclonal antibody e 1:1000 ProteinTech 51064-2-AP MYC tag polyclonal antibody e 1:1000 ProteinTech 16286-1-AP DYKDDDDK tag recombinant antibody (binds to FLAG tag epitope) e 1:500 ProteinTech 80010-1-RR Smooth muscle actin polyclonal antibody IHC: 1:200 IF: 1:100 1:1000 ProteinTech 14395-1-AP Fibronectin monoclonal antibody IHC: 1:200 IF: 1:100 1:2000 ProteinTech 66042-1-Ig SNAIL1 polyclonal antibody e 1:500 ProteinTech 13099-1-AP PAI-1 polyclonal antibody e 1:1000 ProteinTech 13801-1-AP PCSK6 polyclonal antibody IHC: 1:100 1:500 Novus Biologicals (Centennial, CO) NBP1-05030 Collagen type I polyclonal antibody e 1:2000 ProteinTech 14695-1-AP CD31 IHC: 1:100 IF: 1:100 e Santa Cruz Biotechnology (Dallas, TX) sc-376764 PDGFRb IHC: 1:100 IF: 1:100 e Affinity Biosciences (China) AF6133 e, not applicable; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HA, hemagglutinin; IF, immunofluorescence; IHC, immunohistochemical; PAI-1, plasminogen activator inhibitor 1; PCSK6, proprotein convertase subtilisin/kexin-6; PDGFRb, platelet-derived growth factor receptor b; WB, Western blot. at 2 ng/mL for different periods of time, except as otherwise indicated in some experiments.

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining