Collagen I (acid-soluble from rat tail; C-7661) was from Sigma. N-Ethylmaleimide (NEM) was purchased from Sigma (E-1271). The antibodies (Abs) and their sources were as follows: rabbit-anti-DDR1 (SC-532) from Santa Cruz Biotechnology; mouse anti-phosphotyrosine, clone 4G10, from Upstate Biotechnology; rabbit anti-DDR1 (phosphotyrosine 513) from Abbexa Ltd. (Cambridge, UK); goat anti-rabbit Ig horseradish peroxidase-conjugated (P0448, DAKO A/S, Denmark); sheep anti-mouse Ig-horseradish peroxidase (Amersham Biosciences); and goat anti-mouse IgG FITC-conjugated (F-9006, Sigma). Mouse anti-DDR1 monoclonal Abs were generated in our laboratory (19 (link)). Anti-DDR1 (phosphotyrosine 513), here referred to as anti-pY513, is a phospho-specific Ab that recognizes phosphorylated Tyr-513 in DDR1b. Specificity for phosphorylated DDR1b, and not DDR1a, was confirmed (data not shown). Rabbit anti-pY-DDR1 (pY792/796/797), a custom-made Ab against the triple-phosphorylated DDR1 activation loop, was made by Abnova Taiwan Corporation, using as immunogen a peptide encompassing amino acids 780–799 of DDR1 (DDR1b nomenclature). The antigen was KLH-CIKIADFGMSRNL(pY)AGD(pY)(pY)RV.
Free full text: Click here
