Human, Mouse, Lambs, Primate - Macaca fascicularis (Crab-eating Monkey or Cynomolgus Macaque), Primate - Macaca mulatta (Rhesus Macaque), Primates, Rabbit, Xenograft

Applications

Validated:

Immunohistochemistry, Western Blot

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry, Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all LYVE-1 Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human LYVE-1
Ser24-Thr238
Accession # Q9Y5Y7

Specificity

Detects human LYVE-1 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 35% cross-reactivity with recombinant mouse LYVE-1 and less than 1% cross-reactivity with recombinant human CD44 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.30 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human LYVE-1 Antibody

LYVE-1 in Human Tonsil.

LYVE-1 was detected in immersion fixed paraffin-embedded sections of human tonsil using 15 µg/mL Goat Anti-Human LYVE-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2089) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human LYVE-1 by Western Blot.

Western blot shows lysates of human liver and spleen tissue. PVDF membrane was probed with 0.25-1 µg/mL of Goat Anti-Human LYVE-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2089) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for LYVE-1 at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Detection of Human LYVE-1 by Immunocytochemistry/Immunofluorescence

Overview of Multi-dimensional Microscopic Molecular Profiling (MMMP).The overall MMMP approach is depicted using an example tissue section from normal human duodenum (sample #1.9.7). (a) Slides were subjected to repeated cycles of staining and imaging with fluorescent primary antibodies and DAPI. At the end of each cycle, fluorescent signal was removed by a chemical bleaching process, and slides were again imaged, before proceeding to the next round of this iterative procedure. After the final antibody stain (#15 Sma), slides were analyzed with a series of histochemical stains. (b) A set of tiling images spanning each tissue section was initially generated by the microscope system. The tiling images were then computationally ‘stitched’ together to produce a single image per staining cycle for each sample. (c) Image registration was performed to align images from the same tissue section across cycles. Mean intensities of the DAPI signal from all immuno-fluorescence images are shown from before (Unregistered) and after (Registered) the image registration procedure was completed. (d) Following registration, signal intensities from the relevant channels for each image (columns) in the MMMP series were extracted for each pixel (rows) within the tissue section and compiled into a large data matrix of in situ molecular profiles. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0128975), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human LYVE-1 by Western Blot

TGF-beta 1, -beta 2 and -beta 3 reduce lymphatic marker expression in LECs.Primary human LECs were treated with 10, 20 or 30 ng/ml TGF-beta 1, -beta 2 and -beta 3 for 72 hours (a) or 100 hours (b). Untreated cells served as a control. Lysates were prepared and analysed by Western blot using antibodies specific for Lyve-1, Prox-1, VEGFR-3 or vimentin. Vinculin served as loading control. The experiment was performed twice with equivalent results. For densitometry evaluation, protein bands were analysed using the software ImageJ. Bands for the Prox-1, Lyve-1, vimentin and VEGFR-3 proteins were normalized to the corresponding loading control and are displayed as the expression level relative to the untreated control samples. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0162221), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse LYVE-1 by Immunohistochemistry

Peritumoral LECs upregulate MHC-II, PD-L1, and various co-inhibitory molecules compared to naïve dermal LECs. (A) Schematic of experimental design analyzing naïve dermal LECs in the ear and peritumoral LECs of the same mice. (B) Quantification of MHC-II and PD-L1 positivity in naïve and peritumoral LECs (CD45- podoplanin+ CD31+) at day 7, 14, and 21 post B16F10 tumor inoculation in WT (C57/BL6) mice. (C) Quantification of MHC-II and PD-L1 positivity in naïve and peritumoral LECs at day 14 post E0771 tumor inoculation in WT mice. (D) Quantification of MHC-II and PD-L1 positivity in naïve and peritumoral LECs in MMTV-PyMT mice with spontaneous mammary tumors. (E) Quantification of PD-L2, HVEM, and CD48 positivity in naïve and peritumoral LECs at day 21 post B16F10 tumor inoculation in WT mice. (F) Representative immunofluorescence image stained for LYVE-1 (white) and MHC-II (green) in peritumoral and skin margin lymphatics in WT mice at day 14 post B16F10 inoculation. DAPI nuclear staining in blue. Scale bars, 100 μm (left), 20μm (top right and bottom right). Statistical significance assessed using paired student’s t test on data from at least 3 biological replicates; error represented as SD. *, p < 0.05; **, p < 0.01; ***, p < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36362253), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Applications for Human LYVE-1 Antibody

Application

Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil

Western Blot

0.25-1 µg/mL
Sample: Human liver and spleen tissue

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: LYVE-1

Lymphatic vessel endothelial hyaluronan (HA) receptor-1 (LYVE-1) is a receptor of HA, a linear high molecular weight polymer composed of alternating units of D‑glucuronic acid and N-acetyl-D-glucosamine. HA is found in the extracellular matrix of most animal tissues and in body fluids. It modulates cell behavior and functions during tissue remodeling, development, homeostasis, and disease (1). The turnover of HA (several grams/day in humans) occurs primarily in the lymphatics and liver, the two major clearance systems that catabolize approximately 85% and 15% of HA, respectively (1-3). LYVE-1 shares 41% homology with the other known HA receptor, CD44 (4). The homology between the two proteins increases to 61% within the HA binding domain. The HA binding domain, known as the link module, is a common structural motif found in other HA binding proteins such as link protein, aggrecan and versican (1, 5). Human and mouse LYVE-1 share 69% amino acid sequence identity.

LYVE-1 is primarily expressed on both the luminal and abluminal surfaces of lymphatic vessels (4, 5). In addition, LYVE-1 is also present in normal hepatic blood sinusoidal endothelial cells (6). LYVE-1 mediates the endocytosis of HA and may transport HA from tissue to lymph by transcytosis, delivering HA to lymphatic capillaries for removal and degradation in the regional lymph nodes (5, 7, 8). Because of its restricted expression patterns, LYVE-1, along with other lymphatic proteins such as VEGF R3, podoplanin and the homeobox protein propero-related (Prox-1), constitute a set of markers useful for distinguishing between lymphatic and blood microvasculature (4, 5, 9-11).

References

Knudson, C.B. and W. Knudson (1993) FASEB J. 7:1233.
Evered, D and J. Whelan (1989) Ciba Found. Symp. 143:1.
Laurent, T.C. and J.R.F. Fraser (1992) FASEB J. 6:2397.
Banerji, S. et al. (1999) J. Cell Biol. 144:789.
Prevo, R. et al. (2001) J. Biol. Chem. 276:19420.
Jackson, D.J. et al. (2001)Trends Immunol. 22:317.
Zhou, B. et al. (2000) J. Biol. Chem. 275:37733.
Achen, M. et al. (1998) Proc. Natl. Acad. Sci. USA 95:548.
Breiteneder-Gellef, S. et al. (1999) Am. J. Pathol. 154:385.
Wiggle, J.T. and G. Oliver (1999) Cell 98:769.

Long Name

Lymphatic Vessel Endothelial Hyaluronan Receptor 1

Alternate Names

LYVE1, XLKD1

Entrez Gene IDs

10894 (Human); 114332 (Mouse); 293186 (Rat)

Gene Symbol

LYVE1

UniProt

Q9Y5Y7

Additional LYVE-1 Products

Product Documents for Human LYVE-1 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human LYVE-1 Antibody

For research use only

Citations
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