| Autor: |
Sroka IC; Department of Cell Biology and Anatomy, University of Arizona, Tucson, AZ 85724, USA., Chen ML, Cress AE |
| Jazyk: |
angličtina |
| Zdroj: |
Biochemical and biophysical research communications [Biochem Biophys Res Commun] 2008 Oct 24; Vol. 375 (3), pp. 410-3. Date of Electronic Publication: 2008 Aug 17. |
| DOI: |
10.1016/j.bbrc.2008.08.029 |
| Abstrakt: |
Laminins are glycoproteins expressed in the basement membrane of multiple epithelial tissues. Previously described purification procedures for the human laminin variants laminin-5 (LN-332) and laminin-10 (LN-511) use tissue as starting material and have multiple steps. We demonstrate a two-step laminin immunoaffinity purification method to produce consistent quantities of intact and biologically active LN-332 and LN-511 from human keratinocyte (HaCaT) and human lung carcinoma (A549) cell lines, respectively. The purification of LN-332 and LN-551 was demonstrated by PAGE analysis, silver staining and Western blot analysis. The purification procedure includes instruction on removing a cell adhesion contaminant known as galectin-3 binding protein from purified LN-511. The biological activity of purified laminin was tested in a standard cell adhesion assay and compared to commercially available LN-111. This rapid and reproducible purification method will contribute to understanding the role of LN-332 and LN-511 in cell behavior, signaling, and gene expression. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
|
Full Text from ScienceDirect