Anti lyve 1 antibody

Manufactured by Abcam

Sourced in United States, United Kingdom

Anti-LYVE-1 antibody is a laboratory reagent used to detect and study the LYVE-1 protein, which is a cell surface receptor involved in lymphatic vessel development and function. This antibody can be used in various immunological techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to identify and analyze the LYVE-1 protein in biological samples.

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Lab products found in correlation

Donkey serum, by Merck Group (2 mentions) Pe conjugated anti cd31, by BD (2 mentions) Alexa fluor 488 donkey anti rabbit igg, by Thermo Fisher Scientific (2 mentions) Axio observer d1, by Zeiss (2 mentions) Eclipse ti microscope, by Nikon (1 mentions) Anti cd31, by BD (1 mentions) Ivis spectrum imaging system, by PerkinElmer (1 mentions) Luciferin, by Promega (1 mentions) Diaminobenzidine dab solution, by Agilent Technologies (1 mentions) Anti cd34 antibody, by Agilent Technologies (1 mentions) Meyer s hematoxylin, by Merck Group (1 mentions) Alexa 488 conjugated donkey anti rabbit secondary antibody, by Thermo Fisher Scientific (1 mentions) Hanks balanced salt solution (hbss), by Merck Group (1 mentions) Annexin 5 apc, by BD (1 mentions) Alexa fluor 488 anti rabbit, by Thermo Fisher Scientific (1 mentions) Vectashield slide mounting medium with dapi, by Vector Laboratories (1 mentions) Axio imager z1 microscope, by Zeiss (1 mentions) Paraformaldehyde (pfa), by Boster Bio (1 mentions) Sdf 1, by Thermo Fisher Scientific (1 mentions) Goat serum, by Boster Bio (1 mentions)

Close spelling variants of this product

Anti-mouse LYVE-1 rabbit polyclonal antibody Rabbit anti-LYVE-1 polyclonal antibody LYVE-1 antibody Anti-LYVE-1 LYVE-1

12 protocols using anti lyve 1 antibody

Corneal Flat-Mount Immunostaining Protocol

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Corneal flat-mount immunostaining was performed as previously described.^{16 (link)} Briefly, enucleated eyes were fixed in 4% PFA for 20 minutes at 4°C to avoid overfixation of the cornea, rinsed in PBS, and cut in half. Following removal of the extra tissues including lens and iris, the cornea was cut radially to facilitate flat mounting.
For flat-mount immunostaining to detect blood and lymphatic vessels, the corneas were placed in −20°C 100% methanol for 30 minutes and permeabilized with PBS/1% Triton X-100 for 20 minutes at 4°C, and blocked with 5% donkey serum (Sigma-Aldrich Corp.) for 20 minutes, and then stained with anti-Lyve-1 antibodies (Abcam, Cambridge, MA, USA) overnight at 4°C. Corneas were then washed with PBS/0.1% Tween 20 and then stained with PE-conjugated anti-CD31 (BD Biosciences Phamingen, San Diego, CA, USA; clone MEC 13.3) and AlexaFluor488 donkey anti-rabbit IgG (Invitrogen). Corneas were flat mounted and images were recorded on a Zeiss Axio Observer D1.

Seo S., Chen L., Liu W., Zhao D., Schultz K.M., Sasman A., Liu T., Zhang H.F., Gage P.J, & Kume T. (2017). Foxc1 and Foxc2 in the Neural Crest Are Required for Ocular Anterior Segment Development. Investigative Ophthalmology & Visual Science, 58(3), 1368-1377.

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Preclinical Evaluation of circEHBP1 in Bladder Cancer

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All animal studies were carried out according to our previously published guidelines⁴² (link) and permitted by Sun Yat-sen Memorial Hospital, Sun Yat-sen University. Nude BALB/c mice at 4 to 5 weeks old were obtained from the Experimental Animal Center (Guangzhou, P.R. China). The mice were divided randomly into groups, and 5 × 10⁶ UM-UC-3 cells transfected with circEHBP1-overexpressing vector or negative control (NC) vector were inoculated into the right footpad of each mouse. When the tumor reached 200 mm³, the mice were visualized using the IVIS spectral imaging system; the primary tumor and popliteal LNs were removed and then analyzed by IHC using anti-LYVE-1 antibodies (Abcam, Cambridge, UK). Images were visualized under a Nikon Eclipse Ti microscope (Nikon, Japan). Original images of excised popliteal LNs from the nude mice are shown in Figure S5.

Zhu J., Luo Y., Zhao Y., Kong Y., Zheng H., Li Y., Gao B., Ai L., Huang H., Huang J., Li Z, & Chen C. (2021). circEHBP1 promotes lymphangiogenesis and lymphatic metastasis of bladder cancer via miR-130a-3p/TGFβR1/VEGF-D signaling. Molecular Therapy, 29(5), 1838-1852.

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Corneal Neovascularization Induction in Mice

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Corneal neovascularization in 20- to 24-weeks-old male mice by nylon suturing under microscopy was performed according to the previous study ^{21 (link)}. Under general anesthesia, 10-0 nylon suture was placed intra-stromally 1 mm away from the limbal vessel. Ofloxacin ophthalmic ointment was instilled immediately after suturing. For SDF1 neutralizing experiment, sterilized phosphate buffer saline as control or anti-SDF1 antibody (R&D systems) was administered by subconjunctival injection with 200μg/5μl/eye. Cornea and conjunctiva tissues were excised 5 days after sutured and antibody treatment, and double stained with anti-CD31 (BD Biosciences) and anti-LYVE-1 antibodies (Abcam).

Muramatsu M., Nakagawa S., Osawa T., Toyono T., Uemura A., Kidoya H., Takakura N., Usui T., Ryeom S, & Minami T. (2020). Loss of Down syndrome critical region-1 mediated hypercholesterolemia accelerates corneal opacity via pathological neo-vessel formation. Arteriosclerosis, thrombosis, and vascular biology, 40(10), 2425-2439.

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Corneal Immunostaining and Flat Mounting

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Corneal immunostaining and flat mounting was performed as previously described.^{29 (link)} Briefly, enucleated eyes were fixed in 4% paraformaldehyde (PFA) for one hour at room temperature, rinsed in PBS, and cut in half. The iris was removed from the cornea and the cornea was cut radially to facilitate flat mounting. The corneas were placed in 100% methanol at −20°C overnight before immunostaining.
To detect blood and lymphatic vessels in flat-mount immunostaining, the corneas were permeabilized with 1% Triton X-100/PBS for 20 minute, and blocked with 5% donkey serum (Sigma) for 20 minute, and then stained with anti-Lyve-1 antibodies (Abcam) overnight at 4°C. Corneas were then washed with 0.1% Tween 20/PBS and then stained with PE-conjugated anti-CD31 (BD Biosciences, clone MEC 13.3) and AlexaFluor488 donkey anti-rabbit IgG (Invitrogen). Corneas were flat mounted and images were collected by a Zeiss Axio Observer D1 at 2.5X.

Liu W., Schultz K.M., Zhang K., Sasman A., Gao F., Kume T, & Zhang H.F. (2014). In vivo corneal neovascularization imaging by optical-resolution photoacoustic microscopy. Photoacoustics, 2(2), 81-86.

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Monitoring Gastric Cancer Tumor Growth with Kallistatin

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The gastric cancer cell line HGC-27 (2 × 10 6 ) previously infected by pLenti-CMV-EGFP-linker-Luc-PGK-Puro (OBiO Biotech, Shanghai, China) was inoculated into the foot-pads of the two groups of mice (each containing 10 mice) on day 0. When the tumors in the foot-pads reached a volume of ~100 mm 3 , they were imaged with the IVIS Spectrum Imaging System (Perkin Elmer) after injection of luciferin (Promega), and mice were then randomized into two groups. The rKAL-treated group received an intraperitoneal injection of recombinant kallistatin at 48-h intervals to a total amount of 2.88 mg/kg body mass rKAL; the control group was treated with the same volume of phosphate-buffered saline. Tumor growth was monitored by external measurement in two dimensions. Tumor volume was calculated by the following formula: volume (mm 3 ) = (length × width 2 )/2. 28 days after the first injection, tumors in the foot-pads and popliteal lymph nodes were imaged with the IVIS Spectrum Imaging System. The mice were then killed and tumors were dissected and weighed. Last, tumors and popliteal lymph nodes were enucleated and embedded in paraffin. Serial 5-μm sections were analyzed by green fluorescent protein (GFP) imaging and immunohistochemistry (IHC) with anti-LYVE-1 antibodies (Abcam, Cambridge, MA, USA). The images were captured using an AxioVision Rel.4.6 system (Carl Zeiss, Jena, Germany).

Ma C., Luo C., Yin H., Zhang Y., Xiong W., Zhang T., Gao T., Wang X., Che D., Fang Z., Li L., Xie J., Huang M., Zhu L., Jiang P., Qi W., Zhou T., Yang Z., Wang W., Ma J., Gao G, & Yang X. (2018). Kallistatin inhibits lymphangiogenesis and lymphatic metastasis of gastric cancer by downregulating VEGF-C expression and secretion. Gastric cancer : official journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association, 21(4).

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Immunohistochemical Evaluation of Lymphatic Markers

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Consecutive 3 μm sections were cut from each block, and immunohistochemistry was performed as we described previously. An immunoperoxidase technique was done following antigen retrieval with microwave treatment (95°C) in citrate buffer (pH 6.0) for 45 min. After endogenous peroxidase block by 3% H₂O₂-methanol for 15 min, specimens were rinsed with phosphate-buffered saline (PBS) three times. Anti-Prox1 antibody (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), anti-FOXC2 antibody (Santa Cruz Biotechnology), anti-CD34 antibody (a marker for vascular endothelial cells) (DAKO, Carpinteria, CA, USA), and anti-LYVE-1 antibody (a marker for lymphovascular endothelial cells) (Abcam, Tokyo, Japan) diluted by 0.5 μg/ml were used for primary antibody. After 2 h incubated at room temperature, specimens were rinsed with PBS three times and treated for an hour at room temperature with the secondary antibody peroxidase-conjugated anti-mouse (Medical & Biological Laboratories Co., Ltd., Nagoya, Japan) diluted at 0.5%. The specimens were then rinsed with PBS three times and color-developed with diaminobenzidine (DAB) solution (DAKO). After washing, specimens were counterstained with Meyer’s-hematoxylin (Sigma Chemical Co., St. Louis, MO, USA). Immunostaining of all samples was performed at the same conditions of antibody reaction and DAB exposure.

Sasahira T., Ueda N., Yamamoto K., Kurihara M., Matsushima S., Bhawal U.K., Kirita T, & Kuniyasu H. (2014). Prox1 and FOXC2 Act as Regulators of Lymphangiogenesis and Angiogenesis in Oral Squamous Cell Carcinoma. PLoS ONE, 9(3), e92534.

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Isolation and Analysis of Lymphatic Endothelial Cells

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Cells isolated from E15.5 control (Notch1^flx/flx) and conditional Notch1 mutant (Notch1^flx/flx;Prox1CreER^T2) embryos were stained for Lyve-1 and CD31 and subjected to FACS. Briefly, E15.5 embryos were harvested in Hank’s balanced salt solution (HBSS, Sigma-Aldrich) and then chopped for an overnight digestion with collagenease I/II. The colleagenase-treated cell suspension was incubated with RBC (Red blood cell) lysis buffer (StemCell Technologies). Following centrifugation, cell pellets were incubated with anti-Lyve-1 antibody (Abcam) for 20 min at 4°C. After washing with PBS, the cells were then stained with PE-conjugated anti-CD31 antibody (BD Pharmingen) and Alexa 488-conjugated donkey anti-rabbit secondary antibody (Invitrogen). After gauze filtration with a cell strainer (40μm BD Biosciences) to obtain a single cell suspension, the cells were subjected to staining with Annexin V - APC (BD Pharmingen) to assess cell death in the sorted LECs using Flow Fortessa.

Fatima A., Culver A., Culver F., Liu T., Dietz W.H., Thomson B.R., Hadjantonakis A.K., Quaggin S.E, & Kume T. (2014). Murine Notch1 is required for lymphatic vascular morphogenesis during development. Developmental dynamics : an official publication of the American Association of Anatomists, 243(7), 957-964.

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Immunofluorescence Staining of Tissue Sections

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For immunofluorescence staining of tissue samples 5 µm sections of formalin-fixed and paraffin-embedded tissue blocks were used and treated, as described previously^{14 (link)}. The incubation with the anti-LYVE-1 antibody (Abcam, Cambridge; 1:100 in 2% BSA/TBS) occurred over night at 4 °C and after three washing steps with PBS the secondary antibody Alexa fluor 488 anti-rabbit (Life technologies, Carlsbad, USA; 1:500 in PBS) was added for 1 h at 37 °C. Finally, tissue sections were washed again with PBS and VECTASHIELD^TM Slide Mounting Medium with DAPI (Vector Laboratories Inc., Burlingame, USA) was added for mounting. Photographic documentation was performed using the Axio Imager Zeiss Z1 microscope (Axiovision, Carl Zeiss, Oberkochen, Germany).

de Jel M.M., Schott M., Lamm S., Neuhuber W., Kuphal S, & Bosserhoff A.K. (2019). Loss of CYLD accelerates melanoma development and progression in the Tg(Grm1) melanoma mouse model. Oncogenesis, 8(10), 56.

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Lymphangiogenesis and Angiogenesis Assays

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FITC-conjugated mouse anti-human CD34 and mouse IgG1 K isotype control-FITC were from eBioscience (CA, USA). APC-conjugated mouse anti-human VEGFR-3 and mouse IgG1 isotype control-APC were from R&D systems (USA). Anti-LYVE-1 antibody was from Abcam. FITC-conjugated secondary antibodies were from KeyGEN Biotechnology Company (Nanjing, China).
Ficoll-Hypaque solution was purchased from Haoyang Biotechnology Company (Tianjin, China). Goat serum and 4% paraformaldehyde were from Boster Biotechnology Company (Wuhan, China). VEGF-C and SDF-1 were from PeproTech. Lenvatinib (E7080) was from Selleckchem. AMD3100 was from Sigma-Aldrich. Dulbecco's modified Eagle's medium (DMEM) and fetal bovine serum (FBS) were from Hyclone. Complete growth medium EGM-2MV was from Lonza. 4′,6-Diamidino-2-phenylindole (DAPI) was from Beyotime Biotechnology Company (Shanghai, China). JFK (Lot number 130202) was obtained from Jilin Jinfukang Pharmaceutical Co., Ltd., (Jilin, China).

He H.L., Wang D., Tang J., Zhou X.M., Li J.X, & Xu L. (2016). Jin Fu Kang Oral Liquid Inhibits Lymphatic Endothelial Cells Formation and Migration. Evidence-based Complementary and Alternative Medicine : eCAM, 2016, 3635209.

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Tumor Angiogenesis and Lymphangiogenesis Assessment

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Tumors were fixed, embedded in paraffin and sectioned into 4-μm thick. After deparaffinization and rehydration, sections were blocked and then incubated with anti-CD31 antibody (1:50) (Abcam, MA, USA), anti-LYVE1 antibody (1:200) (Abcam, MA, USA) and anti-CD206 antibody (1:50) (Abcam, MA, USA). A biotinylated goat anti-rabbit antibody and ABC Kit were used. CD31 positive ‘hotspots’ represented blood vessels. LYVE-1 positive ‘hotspots’ represented lymphatic vessels. CD206 positive cells were considered as macrophages. Images were quantified at 40X magnification. The quantitative analysis was made on five independent fields per tumor randomly.

Ying X., Wu Q., Wu X., Zhu Q., Wang X., Jiang L., Chen X, & Wang X. (2016). Epithelial ovarian cancer-secreted exosomal miR-222-3p induces polarization of tumor-associated macrophages. Oncotarget, 7(28), 43076-43087.

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