Endocan/ESM-1 Antibody [Unconjugated]

Endocan/ESM-1 was detected in perfusion fixed frozen sections of mouse embryo (13 d.p.c.) using Goat Anti-Mouse Endocan/ESM-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1999) at 5 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to the retina. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Recombinant Mouse Endocan/ESM‑1 (Catalog #1999‑EC), immobilized onto a microplate, supports the adhesion of the Jurkat human acute T cell leukemia cell line in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Mouse Endocan/ESM‑1 (25 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse Endocan/ESM‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1999). The ND50 is typically 1‑5 µg/mL.

Endothelial changes after pericyte depletion. a–f Maximum intensity projection of confocal images from control and DTRiPC P6 retinas stained for IB4 (red) in combination with VEGF-A a, VEGFR2 b, VEGFR3 c, Tie2 d, Esm1 e, and Dll4 f (all in white), as indicated. Note local increase of VEGFR2, VEGFR3, and Esm1 (arrowheads in b, c, e) but not Tie2 or VEGF-A at the edge of the vessel plexus. Dll4 expression in DTRiPC sprouts is increased in some regions (arrowheads) but absent in others (arrows). Scale bar, 100 µm. g–j Quantitation of VEGF-A immunosignals area and intensity g, signal intensity for VEGFR2 h and VEGFR3 i and proportion of Esm1+ area with respect to vascular area j in the P6 control and DTRiPC angiogenic front. Error bars, s.e.m. p-values, Student’s t-test Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29146905), licensed under a CC-BY license. Not internally tested by R&D Systems.

Vascular alterations after intraocular VEGF-A injection. a Morphology of IB4-stained P6 wild-type retinal vessels at 4 h after administration of human VEGF-A165 (0.5 µl at a concentration of 5 μg μl−1). Note blunt appearance of the vessel front after VEGF-A injection but not for vehicle (PBS) control. Scale bar, 200 µm. b Quantitation of sprouts and filopodia at the front of the P6 vessel plexus after injection of VEGF-A165 or vehicle control. Error bars, s.e.m. p-values, Student’s t-test. c PDGFR beta + (green) pericytes are unaffected by short-term VEGF-A administration, whereas VEGFR2 immunosignals (white) are increased in IB4+ (red) ECs (arrowheads). Images shown correspond to insets in a. Scale bar, 100 µm. d Quantitation of VEGFR2 immunosignals intensity in the peripheral plexus of P6 retinas after injection of VEGF-A165 or vehicle control. Error bars, s.e.m. p-values, Student’s t-test. e Confocal images showing increased Esm1 immunostaining (white) in IB4+ (red) ECs in the peripheral plexus (arrowheads) after VEGF-A injection in P6 pups. Scale bar, 200 µm. f VEGF-A165 injection-mediated increase of Esm1 immunosignals (normalized to IB4+ EC area) in the peripheral capillary plexus but not at the edge of the angiogenic front in comparison to PBS-injected controls at P6. Error bars, s.e.m. p-values, Student’s t-test. NS, not statistically significant. g Short-term VEGF-A165 administration leads to clustering of Erg1+ (green) and IB4+ (red) ECs, as indicated, in thick sprout-like structures of P6 retinas. Panels in the center and on the right (scale bar, 20 µm) show higher magnification of the insets on the left (scale bar, 100 µm). Dashed lines in panels on the right outline IB4+ vessels. h Quantitation of EC density in the leading front vessel and emerging sprouts of the P6 angiogenic front after injection of VEGF-A165 or vehicle control. Error bars, s.e.m. p-values, Student’s t-test Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29146905), licensed under a CC-BY license. Not internally tested by R&D Systems.

Inactivation of Flt1 in PDGFR beta + cells. a Experimental scheme of tamoxifen administration for the generation of Flt1iPC mutants. b P6 control, Flt1iPC/+ and Flt1iPC retinas stained with isolectin B4 (IB4). Dashed circles indicate vessel-covered (yellow) and peripheral avascular (white) areas in the overview pictures (top). Scale bar, 500 µm. c Quantitation of body weight and radial outgrowth of the retinal vasculature in control, Flt1iPC/+ and Flt1iPC P6 pups. Error bars, s.e.m. p-values, one-way ANOVA. NS, not statistically significant. d Confocal images of the IB4-stained P6 control, Flt1iPC/+ and Flt1iPC retinal angiogenic front illustrating differences in sprout number and morphology. Scale bar, 100 µm. e Quantitation of sprouts and filopodia in P6 control, Flt1iPC/+ and Flt1iPC retinas. Error bars, s.e.m. p-values, one-way ANOVA and Tukey’s multiple comparison test. NS, not statistically significant. f Confocal images of IB4 (red), Erg1 (green) and VEGFR2 (white) stained P6 retinas highlighting the accumulation of EC nuclei and enhanced VEGFR2 immunosignals (arrowheads) in Flt1iPC sprouts. Vessels are outlined by dashed lines on the right panel. Scale bar, 100 µm. g Quantitation of EC proliferation (EdU+ Erg1+) at the angiogenic front, EC density in sprouts and leading front vessel and VEGFR2 immunosignals intensity in the angiogenic front of control and Flt1iPC P6 retinas. Error bars, s.e.m. p-values, Student’s t-test. h Esm1 (white) expression (arrowheads) in the angiogenic front (IB4+, red, first two columns) and detection of desmin+ pericytes (green, third column) in P6 control and Flt1iPC retinas. Scale bar, 100 µm. i Quantitation of Esm1+ proportion relative to vascular area (IB4+) in the angiogenic front of control and Flt1iPC P6 retinas. Error bars, s.e.m. p-values, Student’s t-test. j Confocal images of P6 retinas stained for NG2 (green) and IB4 (red) showing no significant changes in pericyte coverage in the front (first two columns) or the remodeling plexus around veins (v) or arteries (a) (last two columns). Scale bar, 100 µm. k, l Quantitation of pericyte coverage k and relative gene expression by qPCR on whole lysates l in control and Flt1iPC P6 retinas. Error bars, s.e.m. p-values, Student’s t-test. NS, not statistically significant Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29146905), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Mu
• Applications: WB, IHC, B/N
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Endocan/ESM-1 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse Endocan/ESM-1
  Trp20-Arg184
  Accession # Q9QYY7
• Specificity: Detects mouse Endocan/ESM-1 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 25% cross-reactivity with recombinant human Endocan is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: ESM1
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Neutralization 1-5 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Endocan/ESM-1 Antibody [Unconjugated]

• Endocan
• endothelial cell-specific molecule 1
• ESM1
• ESM-1
• IGFBP-rp6

Background

Endocan (endothelial cell proteoglycan; also known as endothelial-cell specific molecule-1 [ESM-1]), is a 50 kDa, monomeric, secreted, cysteine-rich proteoglycan identified initially in endothelial cells of the kidney and lung (1). Mouse Endocan is synthesized as a 184 amino acid (aa) precursor that contains a 21 aa signal sequence and a 20 kDa, 163 aa mature region (2). The N-terminal 2/3 of the molecule contains 18 cysteine residues and there are no potential N-linked glycosylation sites. Based on human Endocan, there are at least two potential O-linked glycosylation sites, one of which will likely be utilized on Ser at position # 136 of the mature molecule (3). The posttranslational modification is approximately 30 kDa in size. It consists of a single dermatan sulfate chain that contains 4-O sulfated N-acetyl galactosamine with alpha -iduronate. This chain is suggested to bind HGF and contribute to HGF mitogenic activity (4). Mature mouse Endocan is 96% and 74% aa identical to rat and human Endocan, respectively. In human, there is a potential for an alternate splice variant. It shows a deletion of aa’s 82 through 131, a range which would not remove the dermatan sulfate attachment site (4). It is not known if such a splice form exists in mouse. Endocan is expressed by endothelial cells, adipocytes, bronchial epithelium and distal renal tubular epithelium (1, 5, 6). It is upregulated by TNF-alpha and VEGF, (1, 7) and is known to bind to LFA-1 ( alpha _L beta ₂) on the surface of PBMCs, blocking LFA-1 interaction with ICAM-1 (8). Normal circulating levels of Endocan are approximately 1 ng/mL (6).

1. Lassalle, P. et al. (1996) J. Biol. Chem. 271:20458.
2. Lassalle, P. (1999) Genbank Accession #: Q9QYY7.
3. Bechard, D. et al. (2001) J. Biol. Chem. 276:48341.
4. Aitkenhead, M. et al. (2002) Microvasc. Res. 63:159.
5. Wellner, M. et al. (2003) Horm. Metab. Res. 35:217
6. Bechard, D. et al. (2000) J. Vasc. Res. 37:417.
7. Tsai, J.C. et al. (2002) J. Vasc. Res. 39:148.

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 Endocan/ESM-1 Antibody (AF1999)(20)

Publications using AF1999

• E Lee, EA Lee, E Kong, H Chon, M Llaiqui-Co, CH Park, BY Park, NR Kang, JS Yoo, HS Lee, HS Kim, SH Park, SW Choi, D Vestweber, JH Lee, P Kim, WS Lee, I Kim An agonistic anti-Tie2 antibody suppresses the normal-to-tumor vascular transition in the glioblastoma invasion zone Experimental & Molecular Medicine, 2023-02-24;55(2):470-484. 2023-02-24 [PMID: 36828931] (IHC, Mouse)
• Kuo A, Checa A, Niaudet C et al. Murine endothelial serine palmitoyltransferase 1 (SPTLC1) is required for vascular development and systemic sphingolipid homeostasis eLife 2022-10-05 [PMID: 36197001]
• Kuo A, Checa A, Niaudet C et al. Murine endothelial serine palmitoyltransferase 1 (SPTLC1) is required for vascular development and systemic sphingolipid homeostasis eLife 10/5/2022 [PMID: 36197001]
• JH Bae, MJ Yang, SH Jeong, J Kim, SP Hong, JW Kim, YH Kim, GY Koh Gatekeeping role of Nf2/Merlin in vascular tip EC induction through suppression of VEGFR2 internalization Science Advances, 2022-06-10;8(23):eabn2611. 2022-06-10 [PMID: 35687678] (IHC, Mouse)
• T Tedeschi, K Lee, W Zhu, AA Fawzi Limited hyperoxia-induced proliferative retinopathy: A model of persistent retinal vascular dysfunction, preretinal fibrosis and hyaloidal vascular reprogramming for retinal rescue PLoS ONE, 2022-04-27;17(4):e0267576. 2022-04-27 [PMID: 35476813] (IHC, Mouse)
• Cyrine Ben Dhaou, Kamel Mandi, Mickaël Frye, Angela Acheampong, Ayoub Radi, Benjamin De Becker et al. Chemerin regulates normal angiogenesis and hypoxia-driven neovascularization Angiogenesis 5/1/2022 [PMID: 34524600]
• Georgia Zarkada, Joel P. Howard, Xue Xiao, Hyojin Park, Mathilde Bizou, Severine Leclerc et al. Specialized endothelial tip cells guide neuroretina vascularization and blood-retina-barrier formation Developmental Cell 8/1/2021 [PMID: 34273276]
• VC Brücher, C Egbring, T Plagemann, PI Nedvetsky, V Höffken, H Pavenstädt, N Eter, J Kremerskot, P Heiduschka Lack of WWC2 Protein Leads to Aberrant Angiogenesis in Postnatal Mice International Journal of Molecular Sciences, 2021-05-18;22(10):. 2021-05-18 [PMID: 34070186] (IHC, Mouse)
• TS Malinova, A Angulo-Ura, J Nüchel, M Tauber, MM van der St, V Janssen, A de Haan, AG Groenen, M Tebbens, M Graupera, M Plomann, S Huveneers A junctional PACSIN2/EHD4/MICAL-L1 complex coordinates VE-cadherin trafficking for endothelial migration and angiogenesis Nature Communications, 2021-05-10;12(1):2610. 2021-05-10 [PMID: 33972531] (IHC, Mouse)
• Neil Dani, Rebecca H. Herbst, Cristin McCabe, Gilad S. Green, Karol Kaiser, Joshua P. Head et al. A cellular and spatial map of the choroid plexus across brain ventricles and ages Cell 5/1/2021 [PMID: 33932339]
• R Ichijo, M Kabata, H Kidoya, F Muramatsu, R Ishibashi, K Abe, K Tsutsui, H Kubo, Y Iizuka, S Kitano, H Miyachi, Y Kubota, H Fujiwara, A Sada, T Yamamoto, F Toyoshima Vasculature-driven stem cell population coordinates tissue scaling in dynamic organs Science Advances, 2021-02-10;7(7):. 2021-02-10 [PMID: 33568475] (IHC, Mouse)
• Y Zhang, P Chen, S Cai, J Li, Y Chen Decreased expression of endothelial cell specific molecule-1 in lung tissue in emphysematous mice and stable COPD patients Iranian journal of basic medical sciences, 2020-12-01;23(12):1610-1617. 2020-12-01 [PMID: 33489036] (IHC, Western Blot, Mouse)
• Chen J, Lippo L, Labella R et al. Decreased blood vessel density and endothelial cell subset dynamics during ageing of the endocrine system Onco Targets Ther 11/10/2020 [PMID: 33215738]
• ZL Grant, L Whitehead, VHY Wong, Z He, RY Yan, AR Miles, AV Benest, DO Bates, C Prahst, K Bentley, BV Bui, RC Symons, L Coultas Blocking endothelial apoptosis revascularises the retina in a model of ischemic retinopathy J. Clin. Invest., 2020;0(0):. 2020 [PMID: 32427589] (IHC, Mouse)
• Yanagida K, Engelbrecht E, Niaudet C et al. Sphingosine 1-Phosphate Receptor Signaling Establishes AP-1 Gradients to Allow for Retinal Endothelial Cell Specialization Dev. Cell 3/23/2020 [PMID: 32059774]
• Lisandra Vila Ellis, Margo P. Cain, Vera Hutchison, Per Flodby, Edward D. Crandall, Zea Borok et al. Epithelial Vegfa Specifies a Distinct Endothelial Population in the Mouse Lung Developmental Cell 3/1/2020 [PMID: 32059772]
• Xiaocheng Zhao, Pavel Nedvetsky, Fabio Stanchi, Anne-Clemence Vion, Oliver Popp, Kerstin Zühlke et al. Endothelial PKA activity regulates angiogenesis by limiting autophagy through phosphorylation of ATG16L1 eLife 10/3/2019 [PMID: 31580256]
• T Su, Y Zhong, AM Demetriade, J Shen, A Sui, Y Yao, Y Gao, Y Zhu, X Shen, B Xie Endocan Blockade Suppresses Experimental Ocular Neovascularization in Mice Invest. Ophthalmol. Vis. Sci., 2018-02-01;59(2):930-939. 2018-02-01 [PMID: 29450540] (IHC-Fr, Western Blot, Mouse)
• Yamamoto H, Ehling M, Kato K, Kanai K, van Lessen M, Frye M, Zeuschner D, Nakayama M, Vestweber D, Adams R Integrin beta1 controls VE-cadherin localization and blood vessel stability. Nat Commun, 2015-03-10;6(0):6429. 2015-03-10 [PMID: 25752958] (IHC, Mouse)
• Shin JW, Huggenberger R, Detmar M Transcriptional profiling of VEGF-A and VEGF-C target genes in lymphatic endothelium reveals endothelial-specific molecule-1 as a novel mediator of lymphangiogenesis. Blood, 2008-07-09;112(6):2318-26. 2008-07-09 [PMID: 18614759] (IHC-Fr, Mouse)

Bioinformatics

• Gene Symbol: ESM1
• Uniprot:
  • Q9QYY7()