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Recombinant human vegf

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Recombinant human VEGF is a laboratory-produced protein that corresponds to the naturally occurring Vascular Endothelial Growth Factor (VEGF) found in the human body. VEGF is a key regulator of angiogenesis, the process of new blood vessel formation.

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Lab products found in correlation

Human thrombin, by Merck Group (1 mentions) Anti phosphatase cocktail, by Thermo Fisher Scientific (1 mentions) Egm 2 media, by Lonza (1 mentions) Bca assay, by Thermo Fisher Scientific (1 mentions) Recombinant human tnf α, by R&D Systems (1 mentions) Hpaec, by Lonza (1 mentions) Chemiluminescent substrate, by Cell Signaling Technology (1 mentions) 5 bromo 4 chloro 3 indolyl phosphate nitro blue tetrazolium bcip nbt, by Merck Group (1 mentions) Sodium fluoride naf, by Fujifilm (1 mentions) Dimethyl sulfoxide (dmso), by Fujifilm (1 mentions) 4 2 aminoethyl benzenesulfonyl fluoride hydrochloride aebsf, by Merck Group (1 mentions) Phenylmethylsulfonyl fluoride pmsf, by Merck Group (1 mentions) Bovine serum albumin bsa, by Fujifilm (1 mentions) Thiazolyl blue tetrazolium bromide mtt, by Merck Group (1 mentions) Ripa buffer, by Cell Signaling Technology (1 mentions) Bradford reagent, by Bio-Rad (1 mentions) Apatinib, by Selleck Chemicals (1 mentions) Ml385, by Bio-Techne (1 mentions) Vascular endothelial growth factor (vegf), by Bio-Techne (1 mentions) Acriflavine acf, by Merck Group (1 mentions)

6 protocols using recombinant human vegf

1

Angiopoietin-2 Regulation in Endothelial Cells

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Human pulmonary artery endothelial cells (HPAEC) were purchased from Lonza (Basel, Switzerland) and cultured in EGM2 media (Lonza). When cells in multi-well plate reached 90% confluence, media was changed to basal (EBM2) and supplemented with 20 ng/mL recombinant human TNFα (R&D systems, Minneapolis, MN, USA), 2 u/mL human thrombin (Sigma, St Louis, MS, USA), or 30–100 ng/mL recombinant human VEGF (Sigma). After 24 h incubation, media was collected, and cells were lysed with PBS containing 1% Triton and Pierce antiprotease and antiphosphatase cocktail (Thermofisher Scientific, Waltham, MA, USA). Human Angiopoietin 2 levels in media and cell lysates were determined with human Angpt2 Duo-Set ELISA (R&D systems). Protein levels in cell lysates were determined with bicinchoninic acid (BCA) assay (Thermofisher Scientific).
Bogatcheva N.V, & Coleman M.E. (2021). Concentrated Secretome of Adipose Stromal Cells Limits Influenza A Virus-Induced Lung Injury in Mice. Cells, 10(4), 720.
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2

VEGF-induced Cell Proliferation Assay

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Recombinant human VEGF, thiazolyl blue tetrazolium bromide (MTT), 5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium (BCIP/NBT), phenylmethylsulfonyl fluoride (PMSF), and 4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF) were purchased from Sigma-Aldrich Corporation (St. Louis, MO, USA). RIPA buffer and chemiluminescent substrate were obtained from Cell Signaling Technology, Inc. (Beverly, MA, USA). Dimethyl sulfoxide, sodium fluoride (NaF), and bovine serum albumin (BSA) were from Wako Pure Chemical Industries (Osaka, Japan). Bradford reagent was purchased from Bio-Rad Laboratories, Inc. (Hercules, CA, USA). MRBH, also called MGN-3 or BioBran, was provided by Daiwa Pharmaceutical Co., Ltd. (Tokyo, Japan).
ZHU X., OKUBO A., IGARI N., NINOMIYA K, & EGASHIRA Y. (2016). Modified rice bran hemicellulose inhibits vascular endothelial growth factor-induced angiogenesis in vitro via VEGFR2 and its downstream signaling pathways. Bioscience of Microbiota, Food and Health, 36(2), 45-53.
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3

Measuring Endothelial NO Production

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Supp. Figure S2summarizes our sequential cell culture conditions for studies of NO production
and gene/protein expression. 24-well culture plates were coated with 0.1 mg/mL
human fibronectin (Sigma-Aldrich, St. Louis, MO) by incubation at 37°C
for 30 minutes. HUVECs were cultured at 20,000 cells/well and grown to
confluency. A 1:1 mixture of serum from each individual participant and EGM-2
medium was added to different wells (2–3 wells/participant depending on
the experiment) and incubated for 12 hours. The supernatants were harvested for
measurement of NO. The cells were washed twice with Dulbecco’s PBS, and
fresh EGM-2 (not containing serum samples) was added to each well followed by
stimulation with 50 ng/mL recombinant human VEGF (Sigma-Aldrich) for 30 minutes,
after which supernatants were harvested for measurement of stimulated NO levels.
The amount of NO released by the cells before and after VEGF stimulation was
measured by the chemiluminescence method using an NO analyzer (model 280i,
GE/Zysense, Weddington, NC) and normalized to cell count per well. Similar
experiments were performed by incubating cells in varying dilutions of
e-cigarette aerosol condensates depending on the experiment.
Mohammadi L., Han D.D., Xu F., Huang A., Derakhshandeh R., Rao P., Whitlatch A., Cheng J., Keith R.J., Hamburg N.M., Ganz P., Hellman J., Schick S.F, & Springer M.L. (2022). Chronic e-cigarette use impairs endothelial function on the physiological and cellular levels. Arteriosclerosis, thrombosis, and vascular biology, 42(11), 1333-1350.
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4

Investigating VEGF and Oxidative Stress

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MIO-M1 cells were treated with different concentrations of recombinant human VEGF (Sigma-Aldrich, St. Louis, MO, USA), of H2O2 (Sigma-Aldrich), or with 0.1 µM Apatinib (Selleck Chemicals, Houston, TX, USA), a VEGFR2 inhibitor. Before the treatments, the growth medium was replaced with FBS-free medium. H2O2- and VEGF-treated cultures were also treated with 5 µM ML385 (Bio-Techne, Minneapolis, MN, USA), an inhibitor of Nrf2, and with 5 µM acriflavine (ACF, Sigma-Aldrich), a HIF-1 inhibitor. All treatments were performed for 24 h.
Rossino M.G., Lulli M., Amato R., Cammalleri M., Dal Monte M, & Casini G. (2020). Oxidative Stress Induces a VEGF Autocrine Loop in the Retina: Relevance for Diabetic Retinopathy. Cells, 9(6), 1452.
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5

Quantifying Cell Migration via Scratch Assay

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Cell migration was determined by scratch wound assay. The cell monolayer was scratched in a direction perpendicular to a horizontal line by using a 200-μl pipette tip. Human recombinant VEGF (Sigma; 100 ng/ml)-induced cell migration was used as a positive control. At 24 h after the treatment, the average distance for cells migrating into the wound was measured by using ImageJ and normalized to the vehicle control. Data were generated from four fields of view/experiment in a total of three biological replicates.
Zhang D., Ning J., Ramprasath T., Yu C., Zheng X., Song P., Xie Z, & Zou M.H. (2022). Kynurenine promotes neonatal heart regeneration by stimulating cardiomyocyte proliferation and cardiac angiogenesis. Nature Communications, 13, 6371.
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6

Culturing HUVECs under Normoxia and Hypoxia

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Dimethylsulfoxide (DMSO), fetal bovine serum (FBS), antibiotics (penicillin and streptomycin), trypsin-EDTA, and human recombinant VEGF were obtained from Sigma Aldrich (St. Louis, MO, USA). HUVECs were purchased from Lonza (Basel, Switzerland), cultured in EGM complete medium supplemented with SingleQuots™ (containing hydrocortisone, hEGF, FBS, VEGF, hFGF-B, R3-IGF-1, ascorbic acid, heparin and gentamycin/amphotericin-B, Lonza) and incubated at 37°C and 5% CO2 in normoxia (21% O2) or hypoxia (2.5% O2). The hypoxia was guarantee by the use of the hypoxic station InVivo2 200 (Baker Ruskinn, Sanford, MA, USA). To maintain the exponential growth, cells were divided when they reached 80% of confluence in a 25 cm 2 dish. HUVECs at passage between 3 and 8 were used for the experiments.
Turrini E., Ferruzzi L., Guerrini A., Gotti R., Tacchini M., Teti G., Falconi M., Hrelia P, & Fimognari C. (2015). In vitro anti-angiogenic effects of Hemidesmus indicus in hypoxic and normoxic conditions. Journal of ethnopharmacology, 162.
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