pathscan rtk signaling antibody array (chemiluminescent readout)  (Cell Signaling Technology Inc)

Journal: Clinical Cancer Research

Article Title: Spatial Heterogeneity, Stromal Phenotypes, and Therapeutic Vulnerabilities in Colorectal Cancer Peritoneal Metastasis

doi: 10.1158/1078-0432.CCR-24-3780

Figure Lengend Snippet: Development of A5, a novel PAI-1–neutralizing antibody. A, Graphical illustration of phage screen and identification of PAI-1–neutralizing antibody. B, Recognition of different conformations of recombinant PAI-1 by A5 in Western blotting. C, A5 neutralized PAI-1 function in inhibiting tPA and uPA. D, Competitive binding of A5 with fluorine-containing tiplaxtinin on stable active PAI-1 assessed by 19F-NMR. E, Inhibition of proliferation of PM cells treated with PAI-1–positive ascites in vitro by 150 µg/mL A5 in biological triplicates. Human IgG served as a negative control. F, STAT3 signaling activation upon the exposure to PM ascites was suppressed by A5 as compared with IgG control. G, GSEA comparisons of HALLMARK pathway changes between CRC cells treated with A5 ( n = 2) vs. IgG ( n = 2). Pathways with an FDR adjusted P value of less than 0.05 are opacified. CRC, colorectal cancer; NES, normalized enrichment score.

Article Snippet: Then 25 μg of the protein lysate was used to measure total STAT3 and p-STAT3 (Tyr705) by ELISA using the PathScan Total Stat3 Sandwich ELISA kit (Cell Signaling Technology, 7305C) and the PathScan Phospho-Stat3 (Tyr705) Sandwich ELISA kit (Cell Signaling Technology, 7300C), respectively.

Techniques: Recombinant, Western Blot, Binding Assay, Inhibition, In Vitro, Negative Control, Activation Assay, Control