For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu320 of human COL11A1 protein.
The Extracellular Matrix (ECM) is a complex network of macromolecules that provides structural tissue support to cells in the basement membrane and interstitial matrix. It is composed of many molecules, including proteins, glycoproteins, proteoglycans, and polysaccharides (1,2). One of the major proteins that comprise the ECM, and the human body, is collagen. Collagens are a large family of proteins. They are trimeric molecules composed of three alpha polypeptide chains that form a triple helix structure that is characteristic of all collagens (3). The large family of collagens is divided into three subgroups: the fibrillar collagens, the non-fibril forming collagens, and the fibril-associated collagens. These subgroups differ in their structure and supramolecular assembly (3).
Collagen11A1 (COL11A1) is a minor fibrillar collagen that is not normally expressed at high levels in most normal tissues with the exception of cartilage, where it is expressed in high levels, and some other tissues/ organs, where it is expressed at a lower level (4). However, it has been reported that the expression of this molecule is correlated with advanced tumorigenic disease through meta analysis of data from multiple cancers, including ovarian, colon, breast, and lung (5). Additionally, it has also been associated with epithelial-mesenchymal transition (EMT) and metastasis (6,7). Cancer associated fibroblasts (CAFs) are typically the most abundant cell type in the stroma of many solid tumors. They are thought to contribute to ECM stiffness, which is ultimately thought to contribute to tumor growth and resistance to chemotherapeutic intervention. COL11A1 has been found to be elevated in CAFs and may contribute to chemotherapy resistance (8).
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