>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
<0.10 EU per 1 μg of the protein by the LAL method.
Measured by its ability to promote the expansion of E16 rat liver mononuclear cells in vitro, in the presence of Recombinant Mouse SCF/c‑kit Ligand (Catalog # 455-MC), Recombinant Mouse Thrombopoietin/Tpo (Catalog # 488-TO), and Recombinant Mouse Flt‑3 Ligand (Catalog # 427-FL). The ED50 for this effect is 20-100 ng/mL in the presence of a cross‑linking antibody, Mouse Anti-polyHistidine Monoclonal Antibody (Catalog # MAB050).
Chinese Hamster Ovary cell line, CHO-derived mouse Angiopoietin-like Protein 3/ANGPTL3 protein Ser17-Thr206, with a C-terminal 6-His tag
1 μg/lane of Recombinant Mouse Angiopoietin-like 3 (Catalog # 8344-AN) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing both R and NR bands at 23-37 kDa.
Background: Angiopoietin-like Protein 3/ANGPTL3
Angiopoietin-like Protein 3 (ANGPTL3) is a secreted glycoprotein that is structurally related to the angiopoietins (1). Mature mouse ANGPTL3 contains an N-terminal coiled coil domain and a C-terminal fibrinogen-like domain (2). Within the N-terminal fragment, mouse ANGPTL3 shares 83% and 92% aa sequence identity with human and rat ANGPTL3, respectively. ANGPTL3 is expressed in the liver from early in development through adulthood (2, 3). Full length ANGPTL3 circulates in the plasma as do the proteolytically separated N- and C-terminal fragments containing the coiled coil domain and fibrinogen-like domains, respectively (4, 5). The cleavage of ANGPTL3 by Furin and Proprotein Convertase 5/6 is enhanced by its interaction with the related ANGPTL8 (6, 7). ANGPTL3 is found as 70 kDa, 50 kDa, and 32 kDa species and can form weakly associated noncovalent multimers in vitro (3, 4). ANGPTL3 directly inhibits lipoprotein lipase (LPL) and endothelial lipase (EL), enzymes responsible for hydrolyzing circulating triglycerides and HDL phospholipids (8, 9). This activity requires a putative heparin-binding motif which is N-terminal to the coiled coil domain (4). Proteolytic removal of the fibrinogen-like domain from the N-terminal fragment serves to activate ANGPTL3 and increase its ability to inhibit LPL in vitro and function in vivo [Ono 41804]. ANGPTL3 promotes an increase in circulating triglyceride levels without altering VLDL or HDL secretion or uptake (4, 5, 8). ANGPTL3 knockout mice are hypolipidemic and have elevated LPL activity (10). ANGPTL3 expression in vivo is up-regulated by LXR agonists and down-regulated by insulin, leptin, and agonists of TR beta or PPAR beta (11-14). Dysregulated ANGPTL3 expression and elevated plasma triglyceride levels are characteristic of some strains of obese and diabetic mice (5, 8, 12). ANGPTL3 does not bind Tie1 or Tie2, but its fibrinogen-like domain interacts with Integrin alpha V beta 3 to induce endothelial cell adhesion, migration, and neovascularization (15). ANGPTL3, secreted by fetal liver cells, also promotes the expansion of hematopoietic stem cells (16).
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Entrez Gene IDs:
27329 (Human); 30924 (Mouse)
AGNPT5; Ang-5; angiopoietin 5; Angiopoietin-5; angiopoietin-like 3; Angiopoietin-like Protein 3; angiopoietin-related protein 3; ANGPT5angiopoietin-5; ANGPTL3; FHBL2
This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?
The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.
Have you used Recombinant Mouse Angiopoietin-like 3 (aa 17-220), CF?
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