LYVE-1 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

LYVE-1 (Lymphatic Vessel Endothelial Receptor 1) is a receptor for the extracellular matrix mucopolysaccharide hyaluronan (HA) and is primarily expressed by lymphatic endothelial cells in addition to the sinusoidal endothelium of the liver and spleen (1-3). HA, also called hyaluronic acid, is a main component of the extracellular matrix and functions largely in cell adhesion, migration, and tissue remodeling (2). HA undergoes a turnover process that involves release from the tissues to the afferent lymph, degradation within the lymph nodes, and removal of fragments by the liver (2,3). LYVE-1, in addition to other lymphatic proteins including VEGFR3, Prox1, and podoplanin, is a common marker for differentiating between the blood and lymphatic systems (2,3). Furthermore, LYVE-1 is closely related to the leukocyte receptor CD44, having ~44% sequence similarity (1-3). Like CD44, the LYVE-1 protein contains an extracellular HA-binding link domain, with N- and C-terminal extensions, located at the end of a glycosylated juxtamembrane domain stalk region, followed by a transmembrane region, and a cytoplasmic tail (1-3). The key features of the HA-binding like molecule are the three disulfide bridges formed by six cysteine residues (1-3). LYVE-1 is synthesized as a protein of 322 amino acids in length with a theoretical molecular weight of 35 kDa, although due to O-glycosylation often appears in SDS-PAGE at a molecular weight ranging from ~60-70 kDa (2,4). The role of HA-binding is further elucidated by the identification of LYVE-1 as a docking receptor for dendritic cells and macrophages, binding their surface HA to control the entry and migration into lymph vessels (1).

LYVE-1 has been an important marker in studies of embryonic and tumor lymphangiogenesis, as many cancers are characterized by early metastasis to the lymph nodes (1-3, 5). One study of five different vascular tumors in infants used immunohistochemical analysis and found positive LYVE-1 expression in infantile hemangioma, tufted angioma, and kaposiform hemangioendothelioma (5). LYVE-1 along with other markers such as GLUT-1, CD31, CD34, Prox-1, and WT-1 can be used to help provide immunohistologic profiles of various tumors and, when used in conjunction with clinical and histopathologic approaches, may offer better overall diagnosis and disease treatment (5).

References

1. Jackson D. G. (2019). Hyaluronan in the lymphatics: The key role of the hyaluronan receptor LYVE-1 in leucocyte trafficking. Matrix Biology : Journal of the International Society for Matrix Biology. https://doi.org/10.1016/j.matbio.2018.02.001

2. Jackson D. G. (2004). Biology of the lymphatic marker LYVE-1 and applications in research into lymphatic trafficking and lymphangiogenesis. APMIS : acta pathologica, microbiologica, et immunologica Scandinavica. https://doi.org/10.1111/j.1600-0463.2004.apm11207-0811.x

3. Jackson D. G. (2003). The lymphatics revisited: new perspectives from the hyaluronan receptor LYVE-1. Trends in Cardiovascular Medicine. https://doi.org/10.1016/s1050-1738(02)00189-5

4. Unitprot (Q9Y5Y7)

5. Johnson, E. F., Davis, D. M., Tollefson, M. M., Fritchie, K., & Gibson, L. E. (2018). Vascular Tumors in Infants: Case Report and Review of Clinical, Histopathologic, and Immunohistochemical Characteristics of Infantile Hemangioma, Pyogenic Granuloma, Noninvoluting Congenital Hemangioma, Tufted Angioma, and Kaposiform Hemangioendothelioma. The American Journal of Dermatopathology. https://doi.org/10.1097/DAD.0000000000000983

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBL1-12770

• LYVE-1 Antibodies
• LYVE-1 Antibody Pack
• LYVE-1 Lysate
• LYVE-1 Proteins
• LYVE-1 Proteins and Enzyme
• LYVE-1 ELISA