• Purity

  >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie? Blue Staining.

• Endotoxin Level

  <1.0 EU per 1 μg of the protein by the LAL method.  

• Activity

  Measured by the ability of the immobilized protein to support the adhesion of SVEC4?10 mouse vascular endothelial cells. When 5 x 10    ⁴ cells/well are added to Recombinant Human MFG?E8 coated plates (10 μg/mL, 100 μL/well), approximately 45?75% will adhere after 60 minutes at 37 °C.

• Source

  Mouse myeloma cell line, NS0-derived Leu24-Cys387, with a C-terminal 6-His tag

• Accession #

• N-terminal Sequence    
  Analysis

  Leu24

• Predicted Molecular Mass

  41.6 kDa

• SDS-PAGE

  45-50 kDa, reducing conditions

Background: MFG-E8

Milk Fat Globulin Protein E8 (MFG-E8), also known as Lactadherin, MP47, breast epithelial antigen BA46, and SED1, is a 66-75 kDa pleiotropic secreted glycoprotein that promotes mammary gland morphogenesis, angiogenesis, and tumor progression. MFG-E8 also plays an important role in tissue homeostasis and the prevention of inflammation (1). Human MGF-E8 contains one N-terminal EGF-like domain and two C?terminal F5/8-type discoidin-like domains (2). It shares 63% and 61% aa sequence identity with comparable regions of mouse and rat MFG-E8, respectively. Shorter isoforms of human MFG-E8 may have N-terminal deletions (beginning near the end of the first discoidin-like domain), internal deletions (lacking either the EGF-like domain or the central region of the second discoidin-like domain), or C?terminal deletions (truncated within the second discoidin-like domain) (3). A 50 aa internal proteolytic fragment of human MFG-E8 (known as Medin) is a major component of aortic medial amyloid deposits (4). MFG-E8 is released into the milk in complex with lipid-containing milk fat globules. It is also found in multiple other cell types including endothelial cells and smooth muscle cells of the vasculature, immature dendritic cells, at the acrosomal cap of testicular and epididymal sperm, and in epithelial cells of the endometrium (1). MFG-E8 binds to the Integrins alpha V beta 3 and alpha V beta 5 and potentiates the angiogenic action of VEGF through VEGF R2 (5, 6). It reduces inflammation and tissue damage in a variety of settings. MFG-E8 functions as a bridge between phosphatidylserine on apoptotic cells and Integrin alpha V beta 3 on phagocytes, leading to the clearance of apoptotic debris (7). It mediates the engulfment of apoptotic bodies in atherosclerotic plaques and prion-infected brain (8, 9) and of apoptotic B cells during germinal center reactions (10, 11). MFG-E8 also promotes the removal of excess Collagen in fibrotic lungs and the regeneration of damaged intestinal epithelia (12, 13). Its tissue-protective role impairs anti?tumor immunity and chemotherapy-induced apoptosis (14). MFG-E8 in the breastmilk blocks rotavirus infection in nursing babies (15).

• References:

1. Raymond, A. et al. (2009) J. Cell. Biochem. 106:957.

2. Couto, J.R. et al. (1996) DNA Cell Biol. 15:281.

3. Yamaguchi, H. et al. (2010) Eur. J. Immunol. 40:1778.

4. Haggqvist, B. et al. (1999) Proc. Natl. Acad. Sci. USA 96:8669.

5. Silvestre, J.-S. et al. (2005) Nat. Med. 11:499.

6. Borges, E. et al. (2000) J. Biol. Chem. 275:39867.

7. Hanayama, R. et al. (2002) Nature 417:182.

8. Ait-Oufella, H. et al. (2007) Circulation 115:2168.

9. Kranich, J. et al. (2010) J. Exp. Med. 207:2271.

10. Hanayama, R. et al. (2004) Science 304:1147.

11. Kranich, J. et al. (2010) J. Exp. Med. 205:1293.

12. Atabai, K. et al. (2009) J. Clin. Invest. 119:3713.

13. Bu, H.-F. et al. (2007) J. Clin. Invest. 117:3673.

14. Jinushi, M. et al. (2009) J. Exp. Med. 206:1317.

15. Kvistgaard, A.S. et al. (2004) J. Dairy Sci. 87:4088.

• Long Name:

  Milk Fat Globule EGF Factor 8

• Entrez Gene IDs:

  4240 (Human); 17304 (Mouse)

• Alternate Names:

  BA46; Breast epithelial antigen BA46; EDIL1; hP47; HsT19888; lactadherin; Lactahedrin; Medin; MFG1; MFGE8; MFG-E8; MFGM; milk fat globule-EGF factor 8 protein; Milk fat globule-EGF factor 8; O-acetyl disialoganglioside synthase; OAcGD3S; SED1; SPAG10; sperm associated antigen 10; sperm surface protein hP47