Desmocollin-3 is a 100-110kD type I transmembrane glycoprotein that belongs to the cadherin family of calcium dependent adhesion molecules (1-3). The human Desmocollin-3 is synthesized with a 108 amino acid (aa) propeptide. The mature protein consists of a 555 aa extracellular domain (ECD) that contains five cadherin-like domains, a 21 aa transmembrane segment, and a 185 aa cytoplasmic domain (4, 5). Within the ECD, human Desmocollin-3 shares 78% aa sequence identity with mouse and rat Desmocollin-3. It shares 52% and 64% aa sequence identity with Desmocollin-1 and -2, respectively. A splice variant of human Desmocollin-3 has a truncated and substituted cytoplasmic domain (4, 6). Desmocollin-3 is one of the principal components of desmosomes which form adhesive contacts between epithelial cells (1, 2). It is expressed in the basal and suprabasal layers of stratified epithelia in many tissues (5, 7, 8). Desmocollin-1 and -2, by contrast, are preferentially localized in the more superficial layers (7). In mouse, Desmocollin-3 is expressed throughout embryogenesis and is required for non-desmosomal functions during early development (9, 10). During apoptosis, both isoforms of Desmocollin-3 are proteolytically cleaved by caspases and metalloproteases (11). Desmocollin-3 can be upregulated or downregulated in carcinoma (6, 12, 13).
A DNA sequence encoding the extracellular domain of human Desmocollin-3 (DSC-3; Arg 136-Lys 690; Accession # Q14574) was fused to a 6x histidine tag at the C-terminus. The protein was expressed in a mouse myeloma cell line, NSO
Activity:Measured by the ability of the immobilized protein to support the adhesion of BUD-8 human fibroblast cells
When 3x10e4 cells/well are added to rhDSC3 coated plates (10ug/ml, 100ul/well), approximately 45 - 65% will adhere after 90 minutes at 37°C.
Applications:Suitable for use in Western Blot. Other applications not tested.
Recommended Dilution:Western Blot: 62.9kD protein migrates as an ~ 75-85kD protein in SDS-PAGE under reducing conditions. Optimal dilutions to be determined by the researcher.
Storage and Stability:Lyophilized and reconstituted products are stable for 12 months after receipt at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°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.
仕様
Size：50ug
Source Antigen：Recombinant Human, NSO
Grade：Purified
Purity：≥85%, as determined by SDS-PAGE under reducing conditions and visualized by silver stain. Endotoxin: ≤1EU/ug (LAL)
Form：Supplied as a lyophilized powder in PBS. Reconstitute with 500ul sterile PBS.
