Mouse and human VEGF ELISA kits were purchased from Peprotech (900-K99) and R&D System (DEV00), respectively, and assays performed according to manufacturer’s instructions on 100μl of plasma or cell culture media per analysis. For analysis of VEGF protein in muscle, approximately 100mg of frozen gastrocnemius muscle was pulverized using a mortar and pestle, and the powder transferred to a 1.5mL microcentrifuge tube containing 150μL of PBS. The tissue was further disrupted using a mechanical tissue homogenizer (Kimble Kontes Pellet Pestle, Fisher Scientific). After three cycles of freezing and thawing, the tissue suspension was microcentrifuged at max speed for 10min and the supernatant recovered and stored at −80 °C. Lysates were adjusted to 0.5μg/μL in PBS and run using a mouse VEGF ELISA kit from R&D systems (MMV00) was used.
Mouse vegf elisa kit
Manufactured by R&D Systems
Sourced in United States
The Mouse VEGF ELISA kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the measurement of mouse vascular endothelial growth factor (VEGF) levels in cell culture supernatants, serum, and plasma.
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Lab products found in correlation
Mouse il 6 elisa kit, by R&D Systems (2 mentions)
Mouse rat igf 1 elisa kit, by R&D Systems (2 mentions)
Kimble kontes pellet pestle, by Thermo Fisher Scientific (1 mentions)
Vegf human elisa kit, by Thermo Fisher Scientific (1 mentions)
Mouse cxcl12 sdf 1α quantikine elisa kit, by R&D Systems (1 mentions)
Mtt assay, by MP Biomedicals (1 mentions)
96 well plate, by Corning (1 mentions)
Imark microplate reader, by Bio-Rad (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Merck Group (1 mentions)
Protease inhibitor, by Roche (1 mentions)
Phosphatase inhibitor cocktail, by Roche (1 mentions)
Protein assay, by Bio-Rad (1 mentions)
Fulvestrant, by Bio-Techne (1 mentions)
Dacomitinib, by Selleck Chemicals (1 mentions)
Mouse tnf α, by R&D Systems (1 mentions)
Invivopure dilution buffer, by BioXCell (1 mentions)
Rat igg2a clone 2a3, by BioXCell (1 mentions)
17 protocols using mouse vegf elisa kit
1
Quantifying VEGF Protein in Mouse Tissues
2
Quantifying Retinal IL-6 and VEGF
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Lysates were obtained from mouse retinas and centrifuged at 14,000 rpm for 15 min at 4°C. IL-6 and VEGF levels (n = 3, two retinas /experiments) in the lysates were measured using a mouse IL-6 ELISA kit (R&D systems, Minneapolis, MN, USA) and a mouse VEGF ELISA kit (R&D Systems), respectively. ELISA signals were determined using a microplate spectrophotometer (Epoch; BioTek, Winooski, VT, USA).
3
Quantifying VEGF in Cell Media
4
Quantifying VEGF Levels in Serum
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Mouse VEGF ELISA kit (R&D Systems, Minneapolis, MN) was used to detect the concentrations of VEGF in serum according to the manufacturer’s instructions.
5
Quantifying Cytokine Responses in Laser-Induced CNV
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Laser-induced CNV experiments were performed as described above. Four female mice per group were killed and eyes were harvested at days 3 or 5 post laser treatments. Eyes were combined from each animal to maximize protein yield. Whole eye tissue was homogenized and solubilized in ice-cold PBS buffer containing protease inhibitor (catalog No. 11836153001; Roche Biochemicals, Mannheim, Germany). The collected samples at day 3 post laser treatment were assayed for IL-6 protein by using mouse IL-6 ELISA kit (R&D Systems). Samples from day 5 post laser treatment were used for VEGF measurements with the mouse VEGF ELISA kit (R&D Systems).
6
Quantifying VEGF and IGF-1 in CDC Media
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Conditioned media was collected from twice-passaged CDCs 3 days after culture in 8-well chamber culture slides17 (link). A mouse VEGF ELISA kit and a mouse/rat IGF-1 ELISA kit (R&D Systems) were used to measure the levels of VEGF and IGF-1 in the conditioned media30 (link).
7
Retinal VEGF Quantification Assay
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Retinal tissue from the control group and DR model group was added to PBS and homogenized; the homogenate was centrifuged at 12,000 g at 4°C for 10 min. Then, the supernatant was tested for VEGF by mouse VEGF ELISA kit (R&D Systems, MN, U.S.A.). The cells experiments, the cells culture supernatants from the different treatment groups were harvested and tested using mouse VEGF ELISA kit.
8
Quantification of VEGF Protein
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We used a human VEGF ELISA kit (Invitrogen, KHG0111) for HEK293T cell extracts, and a mouse VEGF ELISA kit (R&D Systems, Minneapolis, MN, USA, MMV00) for mouse RPE protein samples, following the manufacturer’s protocol. In short, samples were prepared in a 1:50 dilution and incubated on a precoated plate for 2 h, followed by conjugate and substrate solutions. The optical density was measured using a microplate reader at 450 nm, and VEGFA quantification was calculated against the linear portion of the standard curve. Each sample was run in triplicates.
9
Quantifying VEGF in Cell Media
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To determine the VEGF concentration in the conditioned media, mouse VEGF ELISA kit was used [R&D systems, catalog number: MV000]. After 24-hour treatment, 1mL of the condition media collected from the plates for each treatment was stored at −80°C or quickly analyzed through VEGF ELISA Enzyme-linked immunosorbent assay. The data produced by the ELISA consists of a standard curve and concentrations of VEGF per sample (pg./mL).
10
Quantifying Secreted Factors in CSCs
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Conditioned media was collected from twice-passaged CSCs 3 days after culture in 8-well chamber culture slides. A mouse VEGF ELISA kit and a mouse/rat IGF-1 ELISA kit (R&D Systems) were used to measure the levels of VEGF and IGF-1 in the conditioned medium [18 (link)].
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