protein

DDR1, Fc Chimera, Recombinant, Human (Discoidin Domain Receptor 1, CAK, CD167, EC 2.7.10.1, EDDR1, HGK2, MCK10, NEP, NTRK4, PTK3, PTK3A, RTK6, TRKE)

MBS638129

0.05 mg

710 USD

Recombinant Protein

DDR1, Fc Chimera, Recombinant, Human (Discoidin Domain Receptor 1, CAK, CD167, EC 2.7.10.1, EDDR1, HGK2, MCK10, NEP, NTRK4, PTK3, PTK3A, RTK6, TRKE)

[DDR1, Fc Chimera]

Human

> 85%, as determined by SDS-PAGE and visualized by silver stain.

Supplied as a lyophilized powder from PBS, pH 7.4. Reconstitute with 500ul sterile PBS. For long term storage a carrier protein (e.g. 0.1% HSA or BSA ) should be added to the vial.

Lyophilized powder may be stored at -20 degree C. Stable for 6 months after receipt at -20 degree C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20 degree C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

ELISA (EL/EIA)

Puritiy: Highly Purified. Activity: Measured by its ability to bind Collagen I in a functional ELISA [Leitinger, B. (2003) J. Biol. Chem. 278:16761]. Immobilized Collagen I at 10ug/ml (100ul/well) can bind rhDDR1 with an apparent KD <10nM. Source: Recombinant protein corresponding to Asp21-Thr416 from human DDr1 and Pro 100-Lys330 corresponding to human IgG1 with a IEGRMD linker expressed in mouse NSO cells.

Hybridoma: NSO myeloma cells with spleen cells from Balb/c mice. A DNA sequence encoding the extracellular domain of human DDR1 was fused with the Fc region of human IgG 1 via a linker peptide. The chimeric protein was expressed in a mouse myeloma cell line, NS0. Endotoxin: < 1.0 EU per 1ug. SDS-PAGE: 90-95kD, reducing conditions. Biological Activity: Measured by its ability to bind Collagen I in a functional ELISA. When rhDDR1 is coated at 10ug/ml (100ul/well) rhDDR1 binds with an apparent KD <10nM.

Molecular Biology; MB-Receptors

DDR1, also known as CAK, CD167a, RTK6, and TrkE, is a 120-140kD type I transmembrane glycoprotein that belongs to the discoidin-like domain containing subfamily of receptor tyrosine kinases (1, 2). Mature human DDR2 consists of a 398 amino acid (aa) extracellular domain (ECD) that includes the discoidin-like domain, a 27 aa transmembrane segment, and a 470 aa cytoplasmic region with a tyrosine kinase domain (3). Within the ECD, human DDR1 shares 53% aa sequence identity with human DDR2 and 93% with mouse and rat DDR1. DDR1 is expressed on epithelial tissues, activated monocytes and neutrophils, and in several cancers (2, 4). Compared to isoform DDR1b, DDR1a lacks 37 aa's that include a Shc-interacting NPxY motif in the cytoplasmic juxtamembrane region (5). Two additional kinase deficient splice forms are expressed in colon cancer (6). The discoidin-like domain mediates binding to collagens I-V (1, 7, 8). DDR1 selectively recognizes the triple helical structure of collagen (7, 8). It is expressed on the cell surface as a dimer which can include different isoforms (5, 9). DDR1 oligomerization enhances collagen binding and also modulates collagen fibrillogenesis (10, 11). The transmembrane segment contains a leucine zipper and GxxxG motif, but neither is exclusively required for dimerization (9). Collagen binding induces prolonged autophosphorylation, including the NPxY motif (7, 8). Collagen binding also results in the proteolytic cleavage of a tyrosine phosphorylated 60kD C-terminal fragment (CTF), and a 60kD ECD fragment (12, 13). TIMP3 and TAPI-1 inhibit shedding of the ECD fragment but not the CTF (12). Overexpression of DDR1a promotes MMP-2 activation and results in an increased invasiveness of a glioblastoma cell line; DDR1b does not (14). The recombinant human DDR1/Fc is a disulfide-linked homodimeric protein. Based on N-terminal amino acid sequencing, the recombinant protein starts at Asp 21. Each monomer has a calculated molecular mass of approximately 70.5kD. As a result of glycosylation, recombinant human DDR1 migrates as an approximately 90-95kD protein on a SDS-PAGE under reducing conditions.

0.05 mg

710 USD