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Proteins, Expression, Isolation and Analysis> | The constructs were verified by DNA sequencing. The Fc fusion protein constructs were transfected into Expi293F™ cells using the ExpiFectamine™ Transfection Kit (Gibco). After 7 days, the culture supernatants were collected, and the fusion proteins were purified using affinity chromatography with Protein A-Sepharose beads (GenScript, Piscataway, NJ, USA). After quantifying the protein concentrations using a NanoDrop 2000 (Thermo Fisher Scientific, Waltham, MA, USA) and performing dialysis in phosphate-buffered saline (PBS), the sample purity was analyzed by SDS-PAGE and Coomassie brilliant blue staining. The final pooled fractions were aliquoted and stored at -80°C. | Get A Quote |
Vascular cell adhesion molecule-1 (VCAM-1) is closely associated with tumor progression and metastasis. However, the relevance and role of VCAM-1 in lung cancer have not been clearly elucidated. In this study, we found that VCAM-1 was highly overexpressed in lung cancer tissue compared with that of normal lung, and high VCAM-1 expression correlated with poor survival of lung cancer patients. VCAM-1 knockdown reduced invasion in A549 human lung cancer cells, and competitive blocking experiments targeting the Ig-like domain 6 of VCAM-1 (VCAM-1-D6) demonstrated that the VCAM-1-D6 domain was critical for VCAM-1-mediated A549 cell invasion. Next, we developed a human monoclonal antibody specific to human and mouse V... More