Human MFG-E8 Alexa Fluor® 488-conjugated Antibody

Catalog # Availability Size / Price Qty
IC27671G-100UG
Detection of MFG‑E8 in Human Dendritic Cells by Flow Cytometry
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Human MFG-E8 Alexa Fluor® 488-conjugated Antibody Summary

Species Reactivity
Human
Specificity
Detects human MFG-E8 in direct ELISAs and Western blots.
Source
Monoclonal Mouse IgG2A Clone # 278918
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human MFG-E8
Leu24-Cys387
Accession # Q08431
Formulation
Supplied in a saline solution containing BSA and Sodium Azide.
Label
Alexa Fluor 488 (Excitation= 488 nm, Emission= 515-545 nm)

Applications

Recommended Concentration
Sample
Intracellular Staining by Flow Cytometry
Titration recommended for optimal concentration with starting range of 0.1-1 µg/1 million cells. Sample used for this experiment was Human immature dendritic cells and TF-1 cells fixed with FC012 and permeabilized with FoxP3 Perm.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Intracellular Staining by Flow Cytometry Detection of MFG-E8 antibody in Human Dendritic Cells antibody by Flow Cytometry View Larger

Detection of MFG‑E8 in Human Dendritic Cells by Flow Cytometry Human immature dendritic cells were stained with Mouse Anti-Human MFG-E8 Alexa Fluor® 488-conjugated Monoclonal Antibody (Catalog # IC27671G, filled histogram) or isotype control antibody (IC003G, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). View our protocol for Staining Intracellular Molecules.

Intracellular Staining by Flow Cytometry View Larger

Detection of MFG‑E8 in TF-1 cells by Flow Cytometry. TF-1 cells were stained with Mouse Anti-Human MFG‑E8 Alexa Fluor® 488‑conjugated Monoclonal Antibody (Catalog # IC27671G, filled histogram) or isotype control antibody (Catalog # IC003G, open histogram). To facilitate intracellular staining, cells were fixed with FC012 and permeabilized with FoxP3 Perm. View our protocol for Staining Intracellular Molecules.

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
  • 12 months from date of receipt, 2 to 8 °C as supplied.

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

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Product Specific Notices


This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

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