LYVE-1 Antibody (RM0033-4D17) - Azide and BSA Free

Immunohistochemistry: LYVE-1 Antibody (RM0033-4D17) [NB110-61026] - a-d As revealed by IHC, E12.5 mouse meninges (white bracket) contain cells that co-express LYVE1 (red), PROX1 (green), and MRC1 (white). DAPI marks the nuclei in blue (d). Scale=30 um. n=3 brains. e-h E13.5 mouse meninges (white bracket) contain cells that co-express LYVE1 (red), PROX1 (green), and MRC1 (white). DAPI marks the nuclei in blue (h). Scale=30 um. n=3 brains. i-l E18 mouse meninges (white bracket) contain cells that co-express LYVE1 (red), PROX1 (green), and MRC1 (white). PROX1 is cytoplasmic (j, red arrows) in the meninges but nuclear in neurons (j, blue arrows). DAPI marks the nuclei in blue (l). Scale=30 um. n=3 brains. Image collected and cropped by CiteAb from the following publication (link.springer.com/10.1007/s00401-019-02091-z), licensed under a CC-BY license.

Immunohistochemistry: LYVE-1 Antibody (RM0033-4D17) [NB110-61026] - Cells of human meninges co-express LLEC markers. DAB-IHC with single antibody detecting LYVE1 (b), in the meninges of human post mortem brain showing no signs of neuropathology. These images are taken from a 38 year old male (sample P17/07), and confirmed in n = 2 additional samples. P parenchyma. Scale = 40 um. DAB-IHC with single antibody detecting LYVE1 (c), in elderly human meninges (age: 89-92) with evidence of neuropathology and confirmed in n = 3 brains. P, parenchyma. Scale = 20 um. Image collected and cropped by CiteAb from the following publication (link.springer.com/10.1007/s00401-019-02091-z), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: LYVE1 Antibody (RM0033-4D17) [NB110-61026] - 4% PFA fixed and paraffin embedded mouse Lung and Heart tissue section was subjected to IHC staining of mouse LYVE-1.

Immunocytochemistry/ Immunofluorescence: LYVE-1 Antibody (RM0033-4D17) - Azide and BSA Free [NB110-61026] - Mouse LLECs develop independent of the transcription factor PU.1. a–d As detected by IHC, E15.5 wild-type mice have cells that co-express LYVE1 (a, red) PROX1 (b, green) & MRC1 (c, white) in the developing meninges (white bracket). DAPI labels nuclei blue in the merged image (d). White arrows indicate cells with these three markers. Scale = 50 µm. n = 2 brains. e–h) The meninges (white bracket) of E15.5 PU.1 knockout siblings contain many cells (white arrows) that co-express LYVE1 (e, red), PROX1 (f, green), & MRC1 (g, white). Scale = 50 µm. n = 3 brains Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31696318), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LYVE-1 Antibody (RM0033-4D17) - Azide and BSA Free [NB110-61026] - Mouse LLECs take up A beta 1-40. a Schematic showing the site of dye & A beta 1-40 perfusion into the CSF via the cisterna magna (arrow) of a 2-month old mouse. The dotted line indicates the plane of section. A anterior, P posterior, D dorsal, V ventral. b Coronal brain section indicating the areas imaged. SF4 refers to area captured in Figure S4. c The percentage of each labelled cell type that internalized perfused A beta . Cells co-expressing VEGFR3 & LYVE1 take up A beta at a higher rate than MRC1, LYVE1 double-positive cells as well as MRC1-positive, LYVE1-negative cells (p ≤ 0.05, bootstrap). VEGFR3, LYVE1 counts, n = 2 brains (3 sections/brain). MRC1, LYVE1 counts, n = 3 brains (3 sections/brain). d–d′′′ Cells of the adult mouse meninges that co-express VEGFR3 (d, green) & LYVE1 (d′, white) internalize A beta 1-40 (d′′, cyan). Scale = 20 µm. e-e′′′) Cells of the adult mouse meninges that co-express VEGFR3 (e, green) & MRC1 (e′, white) internalize A beta 1-40 (e′′, cyan). Scale = 40 µm. f–f′′′) Cells of the adult mouse meninges that co-express MRC1 (f, magenta) & LYVE1 (f′, white) internalize A beta 1-40 (f′′, cyan). The walls of a blood vessel (white arrowhead, f′′) also accumulate A beta 1-40. Scale = 60 µm Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31696318), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LYVE-1 Antibody (RM0033-4D17) - Azide and BSA Free [NB110-61026] - Cells with BLEC molecular markers are present within the mouse leptomeninges. a Coronal brain section of adult zebrafish brain indicating the imaging area in the dorsal optic tectum (TeO). b A 14 month old Tg(kdr-l:mCherry); Tg(flt4:mCitrine) double transgenic zebrafish has cells in the meninges (white bracket) that express flt4/vegfr3 ( alpha -GFP, green) near kdr-l positive ( alpha -RFP, red) blood vessels. DAPI (blue) labels the nuclei. Scale = 50 µm. c Coronal mouse brain section showing the imaging areas of the meninges. d As revealed by IHC, 17-week-old mouse brains express VEGFR3 (green) in the meninges (white bracket). Tie2-GFP;NG2-DsRed double reporter mice were used to distinguish arteries & veins. NG2 (red) labels pericytes & smooth muscle cells, Tie2 (magenta) labels vascular endothelial cells, & Hoechst (blue) stains nuclei. The image is rotated with the parenchyma at the bottom for ease of comparison with panel b. Scale = 50 µm. e-e′′′ As revealed by IHC, cells of the meninges co-express MRC1 (e, yellow), LYVE1 (e′, white), & VEGFR3 (e′′, green). Red arrows highlight cells expressing these three markers. The images are rotated with the parenchyma at the bottom. scale = 30 µm. f, g Quantification of the relative numbers of single & double-labelled cells in 2-month old mouse meninges. VEGFR3 & LYVE1 cell counts were from n = 2 brains, 3 coronal sections (10 area images)/brain. MRC1 & LYVE1 cell counts were from n = 3 brains, 3 coronal sections (4 area images)/brain. The mean values for each set are depicted Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31696318), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LYVE-1 Antibody (RM0033-4D17) - Azide and BSA Free [NB110-61026] - Cells of human meninges co-express LLEC markers. a–c DAB-IHC with single antibodies detects VEGFR3 (a), LYVE1 (b), & MRC1 (c) in the meninges of human post mortem brain showing no signs of neuropathology. These images are taken from a 38 year old male (sample P17/07, Table 1), & confirmed in n = 2 additional samples. P parenchyma. Scale = 150 µm (a); 40 µm (b); & 20 µm (c). d–f DAB-IHC with single antibodies detects VEGFR3 (b), LYVE1 (c), & MRC1 (d) in elderly human meninges (age: 89–92) with evidence of neuropathology & confirmed in n = 3 brains (Table 1). P, parenchyma. Scale = 20 µm. g–p IHC with fluorescent antibodies detects human meningeal cells that co-express MRC1 (h, m, yellow), LYVE1 (i, n, white), & VEGFR3 (j, o, green). Nuclei/RNA are labelled with DAPI (g, l, blue) & images are merged in (k, p). Scale = 10 µm Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31696318), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LYVE-1 Antibody (RM0033-4D17) - Azide and BSA Free [NB110-61026] - Cells expressing LLEC markers are present within embryonic mouse leptomeninges. a–d As revealed by IHC, E12.5 mouse meninges (white bracket) contain cells that co-express LYVE1 (red), PROX1 (green), & MRC1 (white). DAPI marks the nuclei in blue (d). Scale = 30 µm. n = 3 brains. e–h E13.5 mouse meninges (white bracket) contain cells that co-express LYVE1 (red), PROX1 (green), & MRC1 (white). DAPI marks the nuclei in blue (h). Scale = 30 µm. n = 3 brains. i–l E18 mouse meninges (white bracket) contain cells that co-express LYVE1 (red), PROX1 (green), & MRC1 (white). PROX1 is cytoplasmic (j, red arrows) in the meninges but nuclear in neurons (j, blue arrows). DAPI marks the nuclei in blue (l). Scale = 30 µm. n = 3 brains Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31696318), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu
• Applications: WB, IHC
• Clone: RM0033-4D17
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated
• Format: Azide and BSA Free
• Concentration: LYOPH

LYVE-1 Antibody (RM0033-4D17) - Azide and BSA Free Summary

• Immunogen: Mouse recombinant LYVE-1
• Marker: Lymphatic Vessel Marker
• Isotype: IgG2
• Clonality: Monoclonal
• Host: Rat
• Gene: LYVE1
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 1:50-1:200
  • Immunohistochemistry-Paraffin 1:50-1:200
  • Western Blot 1:500-1:1000
• Theoretical MW: 35 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: Lyophilized from a 0.2 um filtered solution in PBS. 0.025 mg size is provided in liquid form, PBS
• Preservative: No Preservative
• Concentration: LYOPH
• Purity: Protein G purified
• Reconstitution Instructions: Reconstitute with sterilized PBS to a final concentration of 0.2 mg/ml.

Alternate Names for LYVE-1 Antibody (RM0033-4D17) - Azide and BSA Free

• cell surface retention sequence binding protein-1
• Cell surface retention sequence-binding protein 1
• CRSBP1
• CRSBP-1
• extracellular link domain containing 1
• extracellular link domain-containing 1
• Extracellular link domain-containing protein 1
• HAR
• Hyaluronic acid receptor
• lymphatic vessel endothelial hyaluronan receptor 1
• lymphatic vessel endothelial hyaluronic acid receptor 1
• LYVE1
• LYVE-1
• UNQ230/PRO263
• XLKD1

Background

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

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for LYVE-1 Antibody (NB110-61026)(1)

Publication using NB110-61026

• Shibata-Germanos S, Goodman JR, Grieg A et al. Structural and functional conservation of non-lumenized lymphatic endothelial cells in the mammalian leptomeninges Acta Neuropathol. 2019-11-06 [PMID: 31696318] (IHC-P, Human)

Bioinformatics

• Gene Symbol: LYVE1
• Entrez:
  • 114332(Mouse)
• Uniprot:
  • Q8BHC0()