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Endomucin

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Endomucin is a cell surface glycoprotein that is expressed in endothelial cells. It is a member of the mucin family of proteins.

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Lab products found in correlation

Axio imager, by Zeiss (2 mentions) 780 confocal microscope, by Zeiss (1 mentions) F4 80, by Bio-Rad (1 mentions) α actinin, by Abcam (1 mentions) Smoothened, by Abcam (1 mentions) Cleaved caspse 3, by Cell Signaling Technology (1 mentions) Edu labeling kit, by Thermo Fisher Scientific (1 mentions) Mef2a, by Santa Cruz Biotechnology (1 mentions) Alexa 594, by Jackson ImmunoResearch (1 mentions) Proliferating cell nuclear antigen (pcna), by Santa Cruz Biotechnology (1 mentions) Cyanine3 (cy3), by Jackson ImmunoResearch (1 mentions) Alexa 488, by Jackson ImmunoResearch (1 mentions) Ab106100, by Abcam (1 mentions) Ab22552, by Abcam (1 mentions) Osterix, by Abcam (1 mentions) Prox1, by R&D Systems (1 mentions) Af2727, by R&D Systems (1 mentions) P erk1 2, by Cell Signaling Technology (1 mentions) Mmp 2, by Thermo Fisher Scientific (1 mentions) α sma, by Abcam (1 mentions)

Close spelling variants of this product

Anti-endomucin (2 citations) Rat anti-endomucin (2 citations)

8 protocols using endomucin

1

Quantifying Atherosclerotic Plaque Stability

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To measure the intraplaque contents of smooth muscle cells, macrophages, CD31, endomucin, monocyte chemoattractant protein (MCP)-1, and matrix metalloproteinase (MMP)-2, sections were reacted with primary antibodies against α-smooth muscle actin (α-SMA, Abcam, Cambridge, UK), monocyte/macrophage (MOMA-2, Abcam), CD31 (Abcam), endomucin (Abcam), MCP-1 (Abcam), and MMP-2 (Invitrogen) at 4°C overnight. Then the sections were reacted with secondary antibodies (Zhongshan Golden Bridge Bio-technology, Beijing, China) at 37°C for 30 minutes. The intraplaque macrophages, smooth muscle cells, MMP-2, endomucin, MCP-1and CD31 contents were quantified by Image-Pro Plus 6.0 software. Atherosclerotic plaque instability index was calculated according to the standard formula: (Oil Red O+ area plus MOMA-2+area) / (α-SMA+ area plus collagen+ area) 12 (link)-13 (link).
Luo S., Ma X., Wu W., Lin S., Li M., Zhang Z., Zhu P, & Song Z. (2023). Continuous Hypobaric Hypoxia may Promote Atherosclerosis Progression in Apolipoprotein E-deficient Mice. International Journal of Medical Sciences, 20(7), 849-857.
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2

Immunofluorescence Analysis of Kidney Cells

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Primary antibodies: F4/80 (Bio-Rad, Hercules, CA; clone Cl:A3-1), CD64 (Bio-Rad; clone AT152-9), Endomucin (Abcam; Cambridge, UK; clone V.7C7.1), Ki67 (eBioscience, San Diego, CA; clone SolA15 and Thermo, Waltham, MA; clone SP6), and dsRed (Rockland, Limerick, PA; cat. #600-401-379). Fluorescent-labeled secondary antibodies were used appropriately. 7 μm cryosections were stained using standard protocols. Heat-induced antigen retrieval was performed using pH6.0 sodium citrate solution (eBioscience). Images were captured using Axio imager and 780 confocal microscopes (Zeiss, Oberkochen, Germany). More than three randomly selected areas from 3 to 5 kidneys were imaged and quantified using ImageJ.
Ide S., Yahara Y., Kobayashi Y., Strausser S.A., Ide K., Watwe A., Xu-Vanpala S., Privratsky J.R., Crowley S.D., Shinohara M.L., Alman B.A, & Souma T. (2020). Yolk-sac-derived macrophages progressively expand in the mouse kidney with age. eLife, 9, e51756.
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3

Immunohistochemistry of Tumor Microenvironment

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Immunohistochemistry of TMEM Supplementary Figure 1 for all samples was performed on an autostainer, as described (5 (link), 6 (link), 18 (link)) with only mild modifications. Specifically, the mouse lymphatic vessel TMEM (LV-TMEM) protocol substituted D2-40 for endomucin (1:50 dilution; abcam) to specifically detect lymphatic endothelial cells. The macrophage (IBA-1) and Pan-Mena markers remained the same with our previously described blood vessel TMEM (BV-TMEM) protocol. LV-TMEM and BV-TMEM scores were evaluated and compared by one pathologist in a total of nine metastatic lymph node samples. The scores for both BV-TMEM and LV-TMEM were reported as number of TMEM doorways per 10 high-power fields (HPFs), as described (1 (link), 5 (link), 6 (link)).
Coste A., Karagiannis G.S., Wang Y., Xue E.A., Lin Y., Skobe M., Jones J.G., Oktay M.H., Condeelis J.S, & Entenberg D. (2020). Hematogenous Dissemination of Breast Cancer Cells From Lymph Nodes Is Mediated by Tumor MicroEnvironment of Metastasis Doorways. Frontiers in Oncology, 10, 571100.
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4

Histological Analysis of Animal Tissues

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For histological analysis, animals were euthanized at specified time periods and tissues were fixed in 4% paraformaldehyde. Histological sectioning, hematoxylin-eosin (H&E) staining and Masson Trichome staining were performed as previously described5 (link). Immunohistochemistry was performed on cryosections (10 µm thick) using standard procedures5 (link),50 (link)–52 (link). Briefly, sections were rehydrated, permeabilized, and blocked with 10% normal donkey serum (NDS), 0.1% Triton X-100 in PBS at room temperature and incubated overnight at 4 °C with primary antibodies: α-actinin (Abcam; 1:300), desmin (Novus biologicals; 1:300), Shh (Santa Cruz Biotechnology; 1:200), cleaved caspse-3 (Cell Signaling; 1:300), endomucin (Abcam; 1:100), SM22 (Abcam; 1:400), α-phospho-Histone H3 (Ser10) (Millipore; 1:100), Ki67 (Abcam; 1:200), PCNA (Santa Cruz Biotechnology; 1:100), Mef2a (Santa Cruz Biotechnology; 1:100), Smoothened (Abcam; 1:200), Nkx 2–5 (Santa Cruz Biotechnology; 1:100), and GFP (ThermoFisher Scientific; 1:300) sera. Sections were rinsed and incubated with combinations of secondary antibodies (1:400) including Alexa 488, Alexa 594, Cy3, and Cy5 (Jackson ImmunoResearch Laboratories). EdU staining was performed using the EdU labeling kit (Life Technologies).
Singh B.N., Koyano-Nakagawa N., Gong W., Moskowitz I.P., Weaver C.V., Braunlin E., Das S., van Berlo J.H., Garry M.G, & Garry D.J. (2018). A conserved HH-Gli1-Mycn network regulates heart regeneration from newt to human. Nature Communications, 9, 4237.
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5

Osteoblast and Endothelial Cell Staining

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Tissues were fixed with 4% paraformaldehyde for one to two hours, dehydrated in 30% sucrose overnight, then embedded in OCT without decalcification to preserve antigen activity. Sections were cut at 7 µm. Osterix (Abcam ab22552, Cambridge, MA, USA) and endomucin (Abcam ab106100, Cambridge, MA, USA) were stained as previously described [25 (link)].
Rabjohns E.M., Rampersad R.R., Ghosh A., Hurst K., Eudy A.M., Brozowski J.M., Lee H.H., Ren Y., Mirando A., Gladman J., Bowser J.L., Berg K., Wani S., Ralston S.H., Hilton M.J, & Tarrant T.K. (2023). Aged G Protein-Coupled Receptor Kinase 3 (Grk3)-Deficient Mice Exhibit Enhanced Osteoclastogenesis and Develop Bone Lesions Analogous to Human Paget’s Disease of Bone. Cells, 12(7), 981.
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6

Mouse Eye Wholemount Staining Procedure

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Mouse eye wholemount staining procedure was performed as described elsewhere9 (link). In brief, enucleated mouse eyes were fixed in 2% PFA. After fixation, the eye was trimmed to remove conjunctiva and any remaining connective tissue. A razor blade was used to make an incision from the optic nerve to the center of the cornea. Scissors were used to cut the bottom third of the sclera followed by removal of crystalline lens and retina. The eyes were then blocked and permeabilized in buffer containing 5% donkey serum, 1% BSA and 0.3% Triton X-100. The eyes were then stained with CD31(BD#553370), Prox1(R&D #AF2727), Tie1(R&D #AF619) and Endomucin (Abcam #106100), followed by staining with fluorchrome-conjugated secondary antibodies. After staining, series of small cuts were made using scissors on both the sclera and cornea side toward the limbal region to flatten the SC area. Eyes were mounted so that the out surface of the eye faced the cover slip.
For cross sections of human and mouse eye, eyes were fixed as before and embedded in paraffin for sectioning. 8-um sections were collected on slides and processed for immunofluorescent staining. Stained eyes were imaged by a confocal microscope (Nikon A1). For whole mount eyes, Z stacks across the SC were taken. ImageJ Fiji software10 (link) was used to process and analyze the images.
Du J., Thomson B.R., Onay T, & Quaggin S.E. (2022). Endothelial tyrosine kinase Tie1 is required for normal Schlemm’s canal development. Arteriosclerosis, thrombosis, and vascular biology, 42(3), 348-351.
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7

Histological Analysis of Growth Plate

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Bones were fixed in 10% formalin-PBS (pH 7.4) overnight at 4°C prior to processing for paraffin sectioning. Growth plate morphology was evaluated by Hematoxylin and Eosin (H&E) staining of 5 μM paraffin sections.2 (link),6 (link) Sections were also subjected to immunohistochemistry to evaluate the expression of type-X collagen (1/300, ABclonal, USA), pERK1/2 (1/500, Cell Signaling Technologies), and Endomucin (1/200, Abcam, Waltham, Massachusetts, USA) as previously reported.2 (link),6 (link)
Yadav P.S., Papaioannou G., Kobelski M.M, & Demay M.B. (2023). Phosphate-induced activation of VEGFR2 leads to caspase-9-mediated apoptosis of hypertrophic chondrocytes. iScience, 26(9), 107548.
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8

Immunohistochemical Profiling of Stem Cell Markers

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Information on the staining methods may be found in the literature.1, 2, 3 Tissue sections (5 μm) were deparaffinized and hydrated using standard procedures. Heat‐induced antigen retrieval with citrate buffer pH 6 or Tris‐EDTA pH 9 was done. Primary antibodies against the following proteins: c‐myc (LifeSpan BioSciences, Seattle, WA, USA), Sox2 (GeneTex, Irvine, CA, USA), Nanog (Novus Biologicals, Littleton, CO, USA), VE‐cadherin (Abcam), CD133 (Biorbyt, Cambridge, UK), endomucin (Abcam) and CD31 (Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd, Beijing, China) were applied to the sections. The staining systems used in this study were PicTure PV6000 and Elivision Plus (Zhongshan Chemical Co., Beijing, China).
Zhao X., Sun B., Liu T., Shao B., Sun R., Zhu D., Zhang Y., Gu Q., Dong X., Liu F., Zhao N., Zhang D., Li Y., Meng J., Gong W., Zheng Y, & Zheng X. (2018). Long noncoding RNA n339260 promotes vasculogenic mimicry and cancer stem cell development in hepatocellular carcinoma. Cancer Science, 109(10), 3197-3208.
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