Simplified purification procedure of laminin-332 and laminin-511 from human cell lines.

Biochemical and Biophysical Research Communications
Isis C SrokaAnne E Cress

Abstract

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.

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Citations

May 1, 2010·Journal of Cellular Physiology·Isis C SrokaAnne E Cress
Oct 19, 2012·Cell Adhesion & Migration·Normand Pouliot, Nicole Kusuma
Jan 21, 2014·Biomaterials·Shimon LechtPeter I Lelkes
Nov 13, 2015·Advanced Drug Delivery Reviews·Achilleas D TheocharisNikos K Karamanos
Aug 11, 2016·Journal of Cellular Biochemistry·Lipsa DasAnne E Cress
May 16, 2014·Experimental Dermatology·Xiaoguang LiTakashi Hashimoto
Apr 18, 2014·Molecular Cancer Therapeutics·Terry H LandowskiAnne E Cress

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