Hvegf165

Manufactured by Cell Signaling Technology

Sourced in United States

HVEGF165 is a recombinant human vascular endothelial growth factor 165 (VEGF165) protein produced in human cells. It is a heparin-binding glycoprotein that plays a key role in angiogenesis, the process of new blood vessel formation.

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Lab products found in correlation

Sb216763, by Bio-Techne (1 mentions) Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Primescript first strand cdna synthesis kit, by Takara Bio (1 mentions) Ubiquitin antibody, by Cell Signaling Technology (1 mentions) Sybr green pcr master mix, by Takara Bio (1 mentions) Su5416, by Bio-Techne (1 mentions) Mg132, by Cell Signaling Technology (1 mentions) 12 o tetradecanoylphorbol 13 acetate tpa, by Cell Signaling Technology (1 mentions) Growth factor bullet kit, by Lonza (1 mentions) Huvecs, by Lonza (1 mentions) Proliferating cell nuclear antigen (pcna), by Proteintech (1 mentions) Sodium dodecyl sulfate (sds), by Bio-Rad (1 mentions) Iodoacetamide iaa, by Merck Group (1 mentions) Horseradish peroxidase hrp conjugated secondary antibody, by Beyotime (1 mentions) Dithiothreitol (dtt), by Merck Group (1 mentions) Sodium carbonate, by Merck Group (1 mentions) Ammonium bicarbonate, by Merck Group (1 mentions) Urea, by Bio-Rad (1 mentions)

3 protocols using hvegf165

VEGFR-2 Regulation by GSK-3β and β-TrCP

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Lipofectamine 2000 and TRIzol reagent were obtained from Invitrogen Life Technologies, Inc. (Grand Island, NY, USA). All the primary antibodies [phospho-VEGFR-2 (Tyr1175) (19A10) rabbit monoclonal antibody (mAb) (#2478); VEGFR-2 (D5B1) rabbit mAb (#9698); β-TrCP (D13F10) rabbit mAb (#4394); ubiquitin antibody (#3933); phospho-GSK-3β (Ser9) (D85E12) rabbit mAb (#5558); GSK-3β (D5C5Z) rabbit mAb (#12456)], secondary antibodies [anti-rabbit IgG, HRP-linked antibody (#7074)], human vascular endothelial growth factor 165 (hVEGF₁₆₅) and MG132 (a potent proteasome inhibitor) were purchased from Cell Signaling Technology (Danvers, MA, USA). SU5416 (a specific VEGFR-2 inhibitor) and SB216763 (a specific inhibitor of GSK-3β) was obtained from Tocris Bioscience (Bristol, UK). GO was purchased from Sigma-Aldrich Chemical (St. Louis, USA). β-TrCP siRNA (sc-37178) and lithium chloride (LiCl) were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). LiCl was used to block GSK-3β activity. A protease inhibitor phenylmethylsulfonyl fluoride (PMSF), electro-chemiluminescence (ECL) kit and radio immunoprecipitation assay (RIPA) buffer were purchased from the KeyGen Institute of Biotechnology (Nanjing, China). A PrimeScript First Strand cDNA Synthesis kit and SYBR-Green PCR Master Mix were purchased from Takara Bio (Shiga, Japan).

WU W., ZHANG D., PAN D., ZUO G., REN X, & CHEN S. (2016). Downregulation of vascular endothelial growth factor receptor-2 under oxidative stress conditions is mediated by β-transduction repeat-containing protein via glycogen synthase kinase-3β signaling. International Journal of Molecular Medicine, 37(4), 911-920.

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HUVEC Angiogenesis on Collagen Hydrogels

Cited 2 times

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Human umbilical vein endothelial cells (HUVECs) (Lonza, MD, USA) were cultured in EGM2 basal media supplemented with a growth factor bullet kit (Lonza). HUVECs were seeded at 200cells/mm² on the collagen hydrogel devices between passages 3–4 to yield a rapid monolayer ^{25 (link)}. After formation of a confluent monolayer on the hydrogel surface (~48 hours), hVEGF165 (10ng/ml) (Cell Signaling Technology, MA, USA) and 12-O-Tetradecanoylphorbol-13-Acetate (TPA) (40ng/ml) (Cell Signaling Technology) were added as previously reported ^{17 (link)}. Cell media and growth factors were changed every 48 hours during the experiment. 5mg/ml hydrogel devices were fixed 48 hours after the addition of the growth factors (VEGF and TPA). This time for 7.5mg/ml and 10mg/ml hydrogel devices was increased to 72 hours.

Hosseini Y., Agah M, & Verbridge S.S. (2015). Endothelial Cell Sensing, Restructuring, and Invasion in Collagen Hydrogel Structures. Integrative biology : quantitative biosciences from nano to macro, 7(11), 1432-1441.

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Protein Expression Analysis Protocol

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hVEGF₁₆₅ (Cell Signaling Technology, MA, USA) was diluted with serum-free medium. Antibodies against the following proteins were used: GAPDH (1:1000, Santa Cruz, USA), VEGFA (1:1000, Santa Cruz Biotechnology. Santa Cruz, USA), NRP-1 (1:1000, Abcam, Cambridge, United Kingdom), Nanog (1:2000, Proteintech, Chicago, USA), c-Myc (1:1000, Proteintech, Chicago, USA), CD44 (1:1000, Proteintech, Chicago, USA), CD24 (1:500, Proteintech, Chicago, USA), SOX2 (1:1000, Proteintech, Chicago, USA), OCT-4 (1:1000, Proteintech, Chicago, USA), LEF1 (1:1000, Santa Cruz Biotechnology, Santa Cruz, USA), Cyclin D1 (1:1000, Santa Cruz Biotechnology, Santa Cruz, USA), β-catenin (1:1000, Proteintech, Chicago, USA), PCNA (1:1000, Proteintech, Chicago, USA), GAPVD1 (1:5000, Bethyl Laboratories, Texas, USA) and horseradish peroxidase (HRP)-conjugated secondary antibody (1:2000, Beyotime Biotechnology, China). Ammonium bicarbonate (Sigma‒Aldrich, St. Louis, USA), dithiothreitol (DTT) (Sigma‒Aldrich, St. Louis, USA), iodoacetamide (IAA) (Sigma‒Aldrich, St. Louis, USA), sodium carbonate (Sigma‒Aldrich, St. Louis, USA), acetonitrile (J. T. Baker, Phillipsburg, USA), urea (Bio-Rad, Hercules, CA), sodium dodecyl sulfate (SDS) (Bio-Rad, Hercules, CA) and trypsin (Promega, Madison, USA) were used.

Wang L., Zhang L., Zhao L., Shao S., Ning Q., Jing X., Zhang Y., Zhao F., Liu X., Gu S., Zhao X, & Luo M. (2024). VEGFA/NRP-1/GAPVD1 axis promotes progression and cancer stemness of triple-negative breast cancer by enhancing tumor cell-macrophage crosstalk. International Journal of Biological Sciences, 20(2), 446-463.

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