vegf human elisa kit  (Thermo Fisher)


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    Name:
    VEGF Human ELISA Kit
    Description:
    The human VEGF A ELISA is an enzyme linked immunosorbent assay for the quantitative detection of human VEGF A One of the key molecules for angiogenesis and for the survival of the endothelium is vascular endothelial growth factor VEGF A It is a specific endothelial cell mitogen and a strong vascular permeability factor VPF VEGF A is a heparin binding glycoprotein secreted as a homodimer of 45 kDa by many different cell types VEGF A also causes vasodilation through the nitiric oxide synthase pathway in endothelial cells and can activate migration in monocytes Many different splice variants of VEGF A have been described but VEGF165 is the most predominant protein and anchors with its heparin binding domain to extracellular matrix and to heparin sulfate Examples where VEGF A plays an important role are psoriasis andrheumatoid arthritis as well as the ovarian hyperstimulation syndrome Diabetic retinopathy is associated with high intraocular levels of VEGF A and inhibition of VEGF A function may result in infertility by blockage of corpus luteum function Direct demonstration of the importance of VEGF A in tumor growth has been achieved using dominant negative VEGF receptors to block in vivo proliferation as well as blocking antibodies to VEGF or to one of the VEGF receptors VEGF A transcription is highly activated by hypoxia and by oncogenes like H ras and several transmembrane tyrosine kinases such as epidermal growth factor receptor and ErbB2 Together these pathways account for a marked upregulation of VEGF A in tumors compared to normal tissues and are often of prognostic importance and relevance Targeting the VEGF signalling may be of major therapeutic importance for many diseases and serves as a basis for the design of future anti angiogenic treatments ConjugateBiotinSuitable Sample Typescell culture supernatant serum plasma EDTA citrate heparin Sample Volume50 µLReported ApplicationELISA
    Catalog Number:
    bms277-2
    Price:
    None
    Applications:
    Protein Assays and Analysis|Protein Biology
    Category:
    Kits and Assays
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    Structured Review

    Thermo Fisher vegf human elisa kit
    Characterisation of RP11 - RPE cells revealed polarity and functional defects. a Schematic of RPE differentiation timeline; b Bright-field images of iPSC-derived RPE: representative examples from at least ten independent experiments, scale bar 100 μm; c Immunostaining for basolateral markers BEST1 and Na + /K + -ATPase: representative images from three independent experiments, scale bar 50 μm; d Correct basolateral distribution of collagen IV (C-IV) and apical MERTK in unaffected control (WT3) but not RP11 RPE cells: representative images from three independent experiments, scale bar 50 μm; e , f <t>ELISA</t> assays for apical and basal secretion of PEDF and <t>VEGF,</t> respectively, in control and RP11 - RPE cells; g Trans-epithelial resistance measurements revealed a significant difference between patient and RP11 - RPE cells; h Reduced phagocytic capacity in RP11 - RPE cells. Statistical significance is calculated for MFI (mean fluorescence intensity) values. e – h Data shown as mean ± SEM, n = 3. Statistical significance of pairwise comparisons is indicated by n.s.: not significant; *** p
    The human VEGF A ELISA is an enzyme linked immunosorbent assay for the quantitative detection of human VEGF A One of the key molecules for angiogenesis and for the survival of the endothelium is vascular endothelial growth factor VEGF A It is a specific endothelial cell mitogen and a strong vascular permeability factor VPF VEGF A is a heparin binding glycoprotein secreted as a homodimer of 45 kDa by many different cell types VEGF A also causes vasodilation through the nitiric oxide synthase pathway in endothelial cells and can activate migration in monocytes Many different splice variants of VEGF A have been described but VEGF165 is the most predominant protein and anchors with its heparin binding domain to extracellular matrix and to heparin sulfate Examples where VEGF A plays an important role are psoriasis andrheumatoid arthritis as well as the ovarian hyperstimulation syndrome Diabetic retinopathy is associated with high intraocular levels of VEGF A and inhibition of VEGF A function may result in infertility by blockage of corpus luteum function Direct demonstration of the importance of VEGF A in tumor growth has been achieved using dominant negative VEGF receptors to block in vivo proliferation as well as blocking antibodies to VEGF or to one of the VEGF receptors VEGF A transcription is highly activated by hypoxia and by oncogenes like H ras and several transmembrane tyrosine kinases such as epidermal growth factor receptor and ErbB2 Together these pathways account for a marked upregulation of VEGF A in tumors compared to normal tissues and are often of prognostic importance and relevance Targeting the VEGF signalling may be of major therapeutic importance for many diseases and serves as a basis for the design of future anti angiogenic treatments ConjugateBiotinSuitable Sample Typescell culture supernatant serum plasma EDTA citrate heparin Sample Volume50 µLReported ApplicationELISA
    https://www.bioz.com/result/vegf human elisa kit/product/Thermo Fisher
    Average 99 stars, based on 59 article reviews
    Price from $9.99 to $1999.99
    vegf human elisa kit - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Disrupted alternative splicing for genes implicated in splicing and ciliogenesis causes PRPF31 retinitis pigmentosa"

    Article Title: Disrupted alternative splicing for genes implicated in splicing and ciliogenesis causes PRPF31 retinitis pigmentosa

    Journal: Nature Communications

    doi: 10.1038/s41467-018-06448-y

    Characterisation of RP11 - RPE cells revealed polarity and functional defects. a Schematic of RPE differentiation timeline; b Bright-field images of iPSC-derived RPE: representative examples from at least ten independent experiments, scale bar 100 μm; c Immunostaining for basolateral markers BEST1 and Na + /K + -ATPase: representative images from three independent experiments, scale bar 50 μm; d Correct basolateral distribution of collagen IV (C-IV) and apical MERTK in unaffected control (WT3) but not RP11 RPE cells: representative images from three independent experiments, scale bar 50 μm; e , f ELISA assays for apical and basal secretion of PEDF and VEGF, respectively, in control and RP11 - RPE cells; g Trans-epithelial resistance measurements revealed a significant difference between patient and RP11 - RPE cells; h Reduced phagocytic capacity in RP11 - RPE cells. Statistical significance is calculated for MFI (mean fluorescence intensity) values. e – h Data shown as mean ± SEM, n = 3. Statistical significance of pairwise comparisons is indicated by n.s.: not significant; *** p
    Figure Legend Snippet: Characterisation of RP11 - RPE cells revealed polarity and functional defects. a Schematic of RPE differentiation timeline; b Bright-field images of iPSC-derived RPE: representative examples from at least ten independent experiments, scale bar 100 μm; c Immunostaining for basolateral markers BEST1 and Na + /K + -ATPase: representative images from three independent experiments, scale bar 50 μm; d Correct basolateral distribution of collagen IV (C-IV) and apical MERTK in unaffected control (WT3) but not RP11 RPE cells: representative images from three independent experiments, scale bar 50 μm; e , f ELISA assays for apical and basal secretion of PEDF and VEGF, respectively, in control and RP11 - RPE cells; g Trans-epithelial resistance measurements revealed a significant difference between patient and RP11 - RPE cells; h Reduced phagocytic capacity in RP11 - RPE cells. Statistical significance is calculated for MFI (mean fluorescence intensity) values. e – h Data shown as mean ± SEM, n = 3. Statistical significance of pairwise comparisons is indicated by n.s.: not significant; *** p

    Techniques Used: Functional Assay, Derivative Assay, Immunostaining, Enzyme-linked Immunosorbent Assay, Fluorescence

    2) Product Images from "A convenient protocol for establishing a human cell culture model of the outer retina."

    Article Title: A convenient protocol for establishing a human cell culture model of the outer retina.

    Journal: F1000Research

    doi: 10.12688/f1000research.15409.1

    Physiological studies of ARPE-19 monolayers cultured on transwell inserts. Trans-epithelial Electrical Resistance (TEER) measurements were obtained from long-term cultures to evaluate effectiveness of the Retinal Pigment Epithelial barrier. [ A ] Measurements were conducted from transwells (n=3) at weekly time intervals after seeding. Values were plotted as a percentage change from the previous measurement which show a gradual increase as junctions form and mature. [ B ] Fluctuations between average weekly TEER were observed prior to week 6 (p=0.009 at 3 weeks, p=0.013 at 4 weeks and p=0.001 at 6 weeks, one-way ANOVA with Tukey’s multiple comparisons) after which a stable value of 40.72 Ω.cm 2 was achieved. Data is presented as mean ± SEM. Next, we quantified polarised secretion of Vascular Endothelial Growth Factor (VEGF) and Pigment Epithelium Derived Factor (PEDF) by ARPE-19 cells. Conditioned media was collected (n=3) after 72 hours and proteins quantified by ELISA. [ C ] The apical compartment was found to contain 0.942 ± 0.035ng/ml of VEGF compared to 2.852 ± 0.145ng/ml in the basal chamber, which was statistically significant (p=0.0002). [ D ] PEDF concentrations in the apical compartment was 16.95 ± 0.72ng/ml compared to 25.05 ± 3.93ng/ml in the basal chamber. There were no significant differences (p= 0.112) although more PEDF was secreted via the basolateral RPE surface. Data is presented as mean ± SEM with statistical comparisons made using the unpaired student’s t-test and sourced in part from material published previously 18 under the Creative Commons licence .
    Figure Legend Snippet: Physiological studies of ARPE-19 monolayers cultured on transwell inserts. Trans-epithelial Electrical Resistance (TEER) measurements were obtained from long-term cultures to evaluate effectiveness of the Retinal Pigment Epithelial barrier. [ A ] Measurements were conducted from transwells (n=3) at weekly time intervals after seeding. Values were plotted as a percentage change from the previous measurement which show a gradual increase as junctions form and mature. [ B ] Fluctuations between average weekly TEER were observed prior to week 6 (p=0.009 at 3 weeks, p=0.013 at 4 weeks and p=0.001 at 6 weeks, one-way ANOVA with Tukey’s multiple comparisons) after which a stable value of 40.72 Ω.cm 2 was achieved. Data is presented as mean ± SEM. Next, we quantified polarised secretion of Vascular Endothelial Growth Factor (VEGF) and Pigment Epithelium Derived Factor (PEDF) by ARPE-19 cells. Conditioned media was collected (n=3) after 72 hours and proteins quantified by ELISA. [ C ] The apical compartment was found to contain 0.942 ± 0.035ng/ml of VEGF compared to 2.852 ± 0.145ng/ml in the basal chamber, which was statistically significant (p=0.0002). [ D ] PEDF concentrations in the apical compartment was 16.95 ± 0.72ng/ml compared to 25.05 ± 3.93ng/ml in the basal chamber. There were no significant differences (p= 0.112) although more PEDF was secreted via the basolateral RPE surface. Data is presented as mean ± SEM with statistical comparisons made using the unpaired student’s t-test and sourced in part from material published previously 18 under the Creative Commons licence .

    Techniques Used: Cell Culture, Derivative Assay, Enzyme-linked Immunosorbent Assay

    3) Product Images from "Blockage of store-operated Ca2+ entry antagonizes Epstein–Barr virus-promoted angiogenesis by inhibiting Ca2+ signaling-regulated VEGF production in nasopharyngeal carcinoma"

    Article Title: Blockage of store-operated Ca2+ entry antagonizes Epstein–Barr virus-promoted angiogenesis by inhibiting Ca2+ signaling-regulated VEGF production in nasopharyngeal carcinoma

    Journal: Cancer Management and Research

    doi: 10.2147/CMAR.S159441

    2-APB blunts EBV-promoted endothelial tube formation mediated by VEGF. Notes: ( A ) Effects of 2-APB at increasing concentrations on EGF-evoked Ca 2+ responses are shown. ( B ) Effects of 2-APB at 20 μmol/L on EGF-induced Ca 2+ release from the ER and the following SOCE were measured in the absence and presence of extracellular Ca 2+ , successively. ( C ) Effects of 2-APB at various concentrations on VEGF-A production upon EGF stimulation were determined by ELISA. ( D ) Effects of 2-APB on endothelial tube formation are shown; scale bar =100 μm. The data are representative of three independent experiments and are presented as the mean ± SEM (* P
    Figure Legend Snippet: 2-APB blunts EBV-promoted endothelial tube formation mediated by VEGF. Notes: ( A ) Effects of 2-APB at increasing concentrations on EGF-evoked Ca 2+ responses are shown. ( B ) Effects of 2-APB at 20 μmol/L on EGF-induced Ca 2+ release from the ER and the following SOCE were measured in the absence and presence of extracellular Ca 2+ , successively. ( C ) Effects of 2-APB at various concentrations on VEGF-A production upon EGF stimulation were determined by ELISA. ( D ) Effects of 2-APB on endothelial tube formation are shown; scale bar =100 μm. The data are representative of three independent experiments and are presented as the mean ± SEM (* P

    Techniques Used: Enzyme-linked Immunosorbent Assay

    EBV infection promotes EGF-stimulated VEGF production and endothelial tube formation. Notes: ( A ) VEGF-A production was determined in the mock-controlled and the EBV-infected CNE2 or HK1 cell-conditioned medium. The amount of VEGF released from the serum-starved cells in the absence or presence of extracellular EGF stimulation was determined by ELISA. ( B ) HUVECs were incubated with the cell-conditioned medium as indicated. Representative photographs of HUVEC tube formation were captured at 6 hours after cell seeding; scale bar =100 μm. The tube formation was quantitatively evaluated by calculating the tube length per standard area in each well (right panel). The data are representative of three independent experiments and are presented as the mean ± SEM (* P
    Figure Legend Snippet: EBV infection promotes EGF-stimulated VEGF production and endothelial tube formation. Notes: ( A ) VEGF-A production was determined in the mock-controlled and the EBV-infected CNE2 or HK1 cell-conditioned medium. The amount of VEGF released from the serum-starved cells in the absence or presence of extracellular EGF stimulation was determined by ELISA. ( B ) HUVECs were incubated with the cell-conditioned medium as indicated. Representative photographs of HUVEC tube formation were captured at 6 hours after cell seeding; scale bar =100 μm. The tube formation was quantitatively evaluated by calculating the tube length per standard area in each well (right panel). The data are representative of three independent experiments and are presented as the mean ± SEM (* P

    Techniques Used: Infection, Enzyme-linked Immunosorbent Assay, Incubation

    4) Product Images from "Ginsenoside-Rb1 Induces ARPE-19 Proliferation and Reduces VEGF Release"

    Article Title: Ginsenoside-Rb1 Induces ARPE-19 Proliferation and Reduces VEGF Release

    Journal: ISRN Ophthalmology

    doi: 10.5402/2011/184295

    Effect of different concentrations of Rb1 on VEGF release with increasing time. Cells were seeded at 20,000 per well on day 0. The cells were treated with 0, 0.25, 2.5, 25, and 250 nM concentrations of Rb1 for 24, 48, and 72 hrs. Conditioned media was collected and ELISA performed for the total protein concentration for each day. The results shown in the graph indicate the mean pg of VEGF per 10,000 viable cells for the respective day ± SE. (a), (b), and (c) are the mean pg of VEGF per 10,000 viable cells when treated with different concentrations of Rb1 during the 0 hr to 72 hr treatment periods ± SE. Asterisks indicate statistical significance of P ≤ 0.05 ( n = 3).
    Figure Legend Snippet: Effect of different concentrations of Rb1 on VEGF release with increasing time. Cells were seeded at 20,000 per well on day 0. The cells were treated with 0, 0.25, 2.5, 25, and 250 nM concentrations of Rb1 for 24, 48, and 72 hrs. Conditioned media was collected and ELISA performed for the total protein concentration for each day. The results shown in the graph indicate the mean pg of VEGF per 10,000 viable cells for the respective day ± SE. (a), (b), and (c) are the mean pg of VEGF per 10,000 viable cells when treated with different concentrations of Rb1 during the 0 hr to 72 hr treatment periods ± SE. Asterisks indicate statistical significance of P ≤ 0.05 ( n = 3).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Protein Concentration

    5) Product Images from "The Adenosine A3 Receptor Regulates Differentiation of Glioblastoma Stem-Like Cells to Endothelial Cells under Hypoxia"

    Article Title: The Adenosine A3 Receptor Regulates Differentiation of Glioblastoma Stem-Like Cells to Endothelial Cells under Hypoxia

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19041228

    Blockade and absence of A 3 AR decreases cell differentiation of glioblastoma stem-like cells to endothelial cells under hypoxia. ( A ) Expression of endothelial cell markers (CD31, CD34, CD144, and vWF) analyzed by Flow Cytometry using the mean fluorescence intensity (M.F.I.) in U87MG GSCs treated with the selective antagonist of A 3 AR (MRS1220; 10 μM) under hypoxia (24 h); ( B ) Graphs represent the percentage of positive cells by flow cytometry for each endothelial cell marker in U87MG GSCs treated with MRS1220 (10 μM) and GSCs A 3 AR knockout (GSCs A3-KO ) under hypoxia (24 h); ( C ) Representative flow cytometry histograms of endothelial cell markers in vehicle vs. MRS1220 treated cells; ( D ) Representative flow cytometry histograms of endothelial cell markers in GSCs A3-KO ; ( E ) VEGF-165 ELISA of the supernatant medium of U87MG GSCs treated with 0.001% DMSO (vehicle), MRS1220 and GSCs A 3 AR knockout (GSCs A3-KO ) under hypoxia (72 h). Graphs represent the mean ± S.D. * p
    Figure Legend Snippet: Blockade and absence of A 3 AR decreases cell differentiation of glioblastoma stem-like cells to endothelial cells under hypoxia. ( A ) Expression of endothelial cell markers (CD31, CD34, CD144, and vWF) analyzed by Flow Cytometry using the mean fluorescence intensity (M.F.I.) in U87MG GSCs treated with the selective antagonist of A 3 AR (MRS1220; 10 μM) under hypoxia (24 h); ( B ) Graphs represent the percentage of positive cells by flow cytometry for each endothelial cell marker in U87MG GSCs treated with MRS1220 (10 μM) and GSCs A 3 AR knockout (GSCs A3-KO ) under hypoxia (24 h); ( C ) Representative flow cytometry histograms of endothelial cell markers in vehicle vs. MRS1220 treated cells; ( D ) Representative flow cytometry histograms of endothelial cell markers in GSCs A3-KO ; ( E ) VEGF-165 ELISA of the supernatant medium of U87MG GSCs treated with 0.001% DMSO (vehicle), MRS1220 and GSCs A 3 AR knockout (GSCs A3-KO ) under hypoxia (72 h). Graphs represent the mean ± S.D. * p

    Techniques Used: Cell Differentiation, Expressing, Flow Cytometry, Cytometry, Fluorescence, Marker, Knock-Out, Enzyme-linked Immunosorbent Assay

    Hypoxia increases Cell Differentiation of Glioblastoma Stem-like Cells to Endothelial Cells. ( A ) Expression of Endothelial cell markers (CD31, CD34, CD144, and vWF) analyzed by Flow Cytometry using the mean fluorescence intensity (M.F.I.) in GSCs under normoxia and hypoxia (24 h); ( B ) Graphs represent the percentage of positive cells measured by Flow Cytometry for each Endothelial cell marker; ( C ) Representative Flow Cytometry histograms of (b); ( D ) VEGF-165 ELISA of the supernatant medium of U87MG GSCs in normoxia and hypoxia by 0, 24, 48 and 72 h. Graphs represent the mean ± S.D. * p
    Figure Legend Snippet: Hypoxia increases Cell Differentiation of Glioblastoma Stem-like Cells to Endothelial Cells. ( A ) Expression of Endothelial cell markers (CD31, CD34, CD144, and vWF) analyzed by Flow Cytometry using the mean fluorescence intensity (M.F.I.) in GSCs under normoxia and hypoxia (24 h); ( B ) Graphs represent the percentage of positive cells measured by Flow Cytometry for each Endothelial cell marker; ( C ) Representative Flow Cytometry histograms of (b); ( D ) VEGF-165 ELISA of the supernatant medium of U87MG GSCs in normoxia and hypoxia by 0, 24, 48 and 72 h. Graphs represent the mean ± S.D. * p

    Techniques Used: Cell Differentiation, Expressing, Flow Cytometry, Cytometry, Fluorescence, Marker, Enzyme-linked Immunosorbent Assay

    6) Product Images from "Critical Functionality Effects from Storage Temperature on Human Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium Cell Suspensions"

    Article Title: Critical Functionality Effects from Storage Temperature on Human Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium Cell Suspensions

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-38065-6

    Metabolic Effects from Varied hiPSC-RPE Cell Suspension Temperatures. ( a ) CellTiter-Blue assay shows relative mitochondrial metabolism of hiPSC-RPE cell suspensions from tube condition after 6 hours preservation as measured by relative light absorbance assay. (n = 6) ( b , c ) Glucose and lactate relative quantitation assays of hiPSC-RPE cell suspensions from tube condition after 6 and 24 hours preservation at various temperatures. Dotted line indicates the amount detected in original stock medium (n = 6). ( d , e ) ELISA detection of VEGF (n = 12) and PEDF (n = 9) concentrations calculated from supernatants of hiPSC-RPE cell suspensions from tube condition or culture plate after 6 and 24 hours preservation at various temperatures. Mean ± SEM are presented. P values ( * p
    Figure Legend Snippet: Metabolic Effects from Varied hiPSC-RPE Cell Suspension Temperatures. ( a ) CellTiter-Blue assay shows relative mitochondrial metabolism of hiPSC-RPE cell suspensions from tube condition after 6 hours preservation as measured by relative light absorbance assay. (n = 6) ( b , c ) Glucose and lactate relative quantitation assays of hiPSC-RPE cell suspensions from tube condition after 6 and 24 hours preservation at various temperatures. Dotted line indicates the amount detected in original stock medium (n = 6). ( d , e ) ELISA detection of VEGF (n = 12) and PEDF (n = 9) concentrations calculated from supernatants of hiPSC-RPE cell suspensions from tube condition or culture plate after 6 and 24 hours preservation at various temperatures. Mean ± SEM are presented. P values ( * p

    Techniques Used: CtB Assay, Preserving, Quantitation Assay, Enzyme-linked Immunosorbent Assay

    Functional Assessment and Cell Signature of Preserved hiPSC-RPE Cell Suspensions. ( a ) hiPSC-RPE cell preservations of 6 and 24 hours period at varied temperatures were plated and then cell proliferation was evaluated by MTS assay each day from 1 to 6 days and compared to control populations (n = 9). ( b , c ) VEGF/PEDF secretion quantified by ELISA assay from varied preservations of hiPSC-RPE cells after 13 days in recovery culture and compared to control (n = 6). ( d ) Detection of RPE genes in hiPSC-RPE cells preserved at 16 °C for various periods. RT-qPCR of human RPE65 , TYROSINASE , and PEDF represent (ΔΔCt to the 0 hour control sample) (n = 3). Mean ± SEM are presented. The one-way ANOVA with Tukey’s post hoc pair-wise comparisons test was performed. ( e ) Epithelial morphology in recovery culture of hiPSC-RPE cells preserved at 16 °C for various periods. Cells were plated into a 24-well CELLstart-coated plate after preservation and cultured for 21 days. Morphology was assessed by phase contrast and immunofluorescence microscopy (red, ZO-1). Scale bars = 50 μm.
    Figure Legend Snippet: Functional Assessment and Cell Signature of Preserved hiPSC-RPE Cell Suspensions. ( a ) hiPSC-RPE cell preservations of 6 and 24 hours period at varied temperatures were plated and then cell proliferation was evaluated by MTS assay each day from 1 to 6 days and compared to control populations (n = 9). ( b , c ) VEGF/PEDF secretion quantified by ELISA assay from varied preservations of hiPSC-RPE cells after 13 days in recovery culture and compared to control (n = 6). ( d ) Detection of RPE genes in hiPSC-RPE cells preserved at 16 °C for various periods. RT-qPCR of human RPE65 , TYROSINASE , and PEDF represent (ΔΔCt to the 0 hour control sample) (n = 3). Mean ± SEM are presented. The one-way ANOVA with Tukey’s post hoc pair-wise comparisons test was performed. ( e ) Epithelial morphology in recovery culture of hiPSC-RPE cells preserved at 16 °C for various periods. Cells were plated into a 24-well CELLstart-coated plate after preservation and cultured for 21 days. Morphology was assessed by phase contrast and immunofluorescence microscopy (red, ZO-1). Scale bars = 50 μm.

    Techniques Used: Functional Assay, MTS Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Preserving, Cell Culture, Immunofluorescence, Microscopy

    7) Product Images from "The Adenosine A3 Receptor Regulates Differentiation of Glioblastoma Stem-Like Cells to Endothelial Cells under Hypoxia"

    Article Title: The Adenosine A3 Receptor Regulates Differentiation of Glioblastoma Stem-Like Cells to Endothelial Cells under Hypoxia

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19041228

    Blockade and absence of A 3 AR decreases cell differentiation of glioblastoma stem-like cells to endothelial cells under hypoxia. ( A ) Expression of endothelial cell markers (CD31, CD34, CD144, and vWF) analyzed by Flow Cytometry using the mean fluorescence intensity (M.F.I.) in U87MG GSCs treated with the selective antagonist of A 3 AR (MRS1220; 10 μM) under hypoxia (24 h); ( B ) Graphs represent the percentage of positive cells by flow cytometry for each endothelial cell marker in U87MG GSCs treated with MRS1220 (10 μM) and GSCs A 3 AR knockout (GSCs A3-KO ) under hypoxia (24 h); ( C ) Representative flow cytometry histograms of endothelial cell markers in vehicle vs. MRS1220 treated cells; ( D ) Representative flow cytometry histograms of endothelial cell markers in GSCs A3-KO ; ( E ) VEGF-165 ELISA of the supernatant medium of U87MG GSCs treated with 0.001% DMSO (vehicle), MRS1220 and GSCs A 3 AR knockout (GSCs A3-KO ) under hypoxia (72 h). Graphs represent the mean ± S.D. * p
    Figure Legend Snippet: Blockade and absence of A 3 AR decreases cell differentiation of glioblastoma stem-like cells to endothelial cells under hypoxia. ( A ) Expression of endothelial cell markers (CD31, CD34, CD144, and vWF) analyzed by Flow Cytometry using the mean fluorescence intensity (M.F.I.) in U87MG GSCs treated with the selective antagonist of A 3 AR (MRS1220; 10 μM) under hypoxia (24 h); ( B ) Graphs represent the percentage of positive cells by flow cytometry for each endothelial cell marker in U87MG GSCs treated with MRS1220 (10 μM) and GSCs A 3 AR knockout (GSCs A3-KO ) under hypoxia (24 h); ( C ) Representative flow cytometry histograms of endothelial cell markers in vehicle vs. MRS1220 treated cells; ( D ) Representative flow cytometry histograms of endothelial cell markers in GSCs A3-KO ; ( E ) VEGF-165 ELISA of the supernatant medium of U87MG GSCs treated with 0.001% DMSO (vehicle), MRS1220 and GSCs A 3 AR knockout (GSCs A3-KO ) under hypoxia (72 h). Graphs represent the mean ± S.D. * p

    Techniques Used: Cell Differentiation, Expressing, Flow Cytometry, Cytometry, Fluorescence, Marker, Knock-Out, Enzyme-linked Immunosorbent Assay

    Hypoxia increases Cell Differentiation of Glioblastoma Stem-like Cells to Endothelial Cells. ( A ) Expression of Endothelial cell markers (CD31, CD34, CD144, and vWF) analyzed by Flow Cytometry using the mean fluorescence intensity (M.F.I.) in GSCs under normoxia and hypoxia (24 h); ( B ) Graphs represent the percentage of positive cells measured by Flow Cytometry for each Endothelial cell marker; ( C ) Representative Flow Cytometry histograms of (b); ( D ) VEGF-165 ELISA of the supernatant medium of U87MG GSCs in normoxia and hypoxia by 0, 24, 48 and 72 h. Graphs represent the mean ± S.D. * p
    Figure Legend Snippet: Hypoxia increases Cell Differentiation of Glioblastoma Stem-like Cells to Endothelial Cells. ( A ) Expression of Endothelial cell markers (CD31, CD34, CD144, and vWF) analyzed by Flow Cytometry using the mean fluorescence intensity (M.F.I.) in GSCs under normoxia and hypoxia (24 h); ( B ) Graphs represent the percentage of positive cells measured by Flow Cytometry for each Endothelial cell marker; ( C ) Representative Flow Cytometry histograms of (b); ( D ) VEGF-165 ELISA of the supernatant medium of U87MG GSCs in normoxia and hypoxia by 0, 24, 48 and 72 h. Graphs represent the mean ± S.D. * p

    Techniques Used: Cell Differentiation, Expressing, Flow Cytometry, Cytometry, Fluorescence, Marker, Enzyme-linked Immunosorbent Assay

    8) Product Images from "The truncated somatostatin receptor sst5TMD4 stimulates the angiogenic process and is associated to lymphatic metastasis and disease-free survival in breast cancer patients"

    Article Title: The truncated somatostatin receptor sst5TMD4 stimulates the angiogenic process and is associated to lymphatic metastasis and disease-free survival in breast cancer patients

    Journal: Oncotarget

    doi: 10.18632/oncotarget.11076

    sst5TMD4 expression is associated to higher expression of pro-angiogenic factors and higher capacity to form mammospheres in breast cancer MCF-7 cells A. Software-driven functional analysis of genes whose expression is altered by the presence of sst5TMD4 in MCF-7 cells by gene expression microarray (green = inhibition, red = overexpression). B. User-driven supervised functional analysis of genes whose expression is altered by the presence of sst5TMD4 in MCF-7 cells by gene expression microarray (green = inhibition, red = overexpression). C. Examples of sst5TMD4-induced gene expression changes validated by additional qPCR in transfected cell lines. D. Changes in the expression of angiogenesis-related genes (VEGF, EGF, Ang1, Ang2, HIF1a and HIF1b) measured by qPCR in MCF-7 cells stably transfected with sst5TMD4 or pCDNA3.1 empty vector (mock). E. Levels of secreted VEGF in MCF-7 cells stably transfected with sst5TMD4 and mock controls measured by ELISA. F. Percentage and representative images of mammospheres formed from MCF-7 cells stably transfected with sst5TMD4 and the respective mock controls. Data represent mean ± SEM of n=3-6 independent experiments. Asterisks (*, p
    Figure Legend Snippet: sst5TMD4 expression is associated to higher expression of pro-angiogenic factors and higher capacity to form mammospheres in breast cancer MCF-7 cells A. Software-driven functional analysis of genes whose expression is altered by the presence of sst5TMD4 in MCF-7 cells by gene expression microarray (green = inhibition, red = overexpression). B. User-driven supervised functional analysis of genes whose expression is altered by the presence of sst5TMD4 in MCF-7 cells by gene expression microarray (green = inhibition, red = overexpression). C. Examples of sst5TMD4-induced gene expression changes validated by additional qPCR in transfected cell lines. D. Changes in the expression of angiogenesis-related genes (VEGF, EGF, Ang1, Ang2, HIF1a and HIF1b) measured by qPCR in MCF-7 cells stably transfected with sst5TMD4 or pCDNA3.1 empty vector (mock). E. Levels of secreted VEGF in MCF-7 cells stably transfected with sst5TMD4 and mock controls measured by ELISA. F. Percentage and representative images of mammospheres formed from MCF-7 cells stably transfected with sst5TMD4 and the respective mock controls. Data represent mean ± SEM of n=3-6 independent experiments. Asterisks (*, p

    Techniques Used: Expressing, Software, Functional Assay, Microarray, Inhibition, Over Expression, Real-time Polymerase Chain Reaction, Transfection, Stable Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay

    9) Product Images from "Transmigration of Neural Stem Cells across the Blood Brain Barrier Induced by Glioma Cells"

    Article Title: Transmigration of Neural Stem Cells across the Blood Brain Barrier Induced by Glioma Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0060655

    HGF, VEGF and the lack of EGF in glioma C6 CM induce NSCs transmigration across RBMECs. A) The amount of HGF and VEGF determined by ELISA is higher in glioma C6 CM than in astrocyte CM, while the opposite is found for EGF. IP of HGF, VEGF or EGF reduces the amount of the growth factor present in CM. HGF: N = 4, F(3,12) = 26.54; VEGF: N = 3, F(3,8) = 20.98; EGF: N = 3, F(3,8) = 44.52; **P
    Figure Legend Snippet: HGF, VEGF and the lack of EGF in glioma C6 CM induce NSCs transmigration across RBMECs. A) The amount of HGF and VEGF determined by ELISA is higher in glioma C6 CM than in astrocyte CM, while the opposite is found for EGF. IP of HGF, VEGF or EGF reduces the amount of the growth factor present in CM. HGF: N = 4, F(3,12) = 26.54; VEGF: N = 3, F(3,8) = 20.98; EGF: N = 3, F(3,8) = 44.52; **P

    Techniques Used: Transmigration Assay, Enzyme-linked Immunosorbent Assay

    10) Product Images from "Physical disruption of cell–cell contact induces VEGF expression in RPE cells"

    Article Title: Physical disruption of cell–cell contact induces VEGF expression in RPE cells

    Journal: Molecular Vision

    doi:

    Micropatterning results. A : Formation of varying concentrations of ARPE-19 cell with a free edge by micropatterning. Black = average number of cells per cell patch of corresponding size; red = concentration of cells losing cell–cell contact at least on one side. Increase in the concentration of cells losing cell–cell contact correlates negatively with cell count and patch size. Data represent the mean ± standard deviation for three replicates (seven measurements per replicate for each pattern size and each time point; n = 21). Similar results were seen for the human RPE (hRPE) cells (data not shown). B : Increase in the cell patch diameter at day 2. A greater increase in patch diameter was observed for the ARPE-19 cells compared with the hRPE cells. Data represent the mean ± standard deviation for three replicates (seven measurements per replicate for each pattern size and each time point; n = 21). C , D : VEGF expression analysis with enzyme-linked immunosorbent assay (ELISA) for the micropatterned ARPE-19 ( C ) and hRPE ( D ) cells. VEGF expression is presented as the total VEGF divided by the cell count. The smaller the pattern size, the higher the VEGF expression per cell. VEGF expression decreased during day 2 presumably because of the increased pattern sizes due to cell growth ( B ). Data represent the mean ± standard deviation for three replicates (three measurements per replicate for each pattern size and each time point; n = 9); * p
    Figure Legend Snippet: Micropatterning results. A : Formation of varying concentrations of ARPE-19 cell with a free edge by micropatterning. Black = average number of cells per cell patch of corresponding size; red = concentration of cells losing cell–cell contact at least on one side. Increase in the concentration of cells losing cell–cell contact correlates negatively with cell count and patch size. Data represent the mean ± standard deviation for three replicates (seven measurements per replicate for each pattern size and each time point; n = 21). Similar results were seen for the human RPE (hRPE) cells (data not shown). B : Increase in the cell patch diameter at day 2. A greater increase in patch diameter was observed for the ARPE-19 cells compared with the hRPE cells. Data represent the mean ± standard deviation for three replicates (seven measurements per replicate for each pattern size and each time point; n = 21). C , D : VEGF expression analysis with enzyme-linked immunosorbent assay (ELISA) for the micropatterned ARPE-19 ( C ) and hRPE ( D ) cells. VEGF expression is presented as the total VEGF divided by the cell count. The smaller the pattern size, the higher the VEGF expression per cell. VEGF expression decreased during day 2 presumably because of the increased pattern sizes due to cell growth ( B ). Data represent the mean ± standard deviation for three replicates (three measurements per replicate for each pattern size and each time point; n = 9); * p

    Techniques Used: Concentration Assay, Cell Counting, Standard Deviation, Expressing, Enzyme-linked Immunosorbent Assay

    11) Product Images from "All-trans retinoic acid impairs the vasculogenic mimicry formation ability of U87 stem-like cells through promoting differentiation"

    Article Title: All-trans retinoic acid impairs the vasculogenic mimicry formation ability of U87 stem-like cells through promoting differentiation

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2015.3449

    VEGF transcript and protein expression in U87 glioblastoma SLCs incubated with different concentrations of ATRA. (A) Reverse transcription quantitative polymerase chain reaction was used to determine the expression of VEGF transcripts in different treatment groups (BC; NC; and 1, 10 and 100 nmol/l ATRA). GAPDH was used as an internal control. (B) Enzyme-linked immunosorbent assay analysis was used to determine protein expression levels of VEGF in different treatment groups (BC; NC; and 1, 10 and 100 nmol/l ATRA). Values are presented as the mean ± standard deviation. * P
    Figure Legend Snippet: VEGF transcript and protein expression in U87 glioblastoma SLCs incubated with different concentrations of ATRA. (A) Reverse transcription quantitative polymerase chain reaction was used to determine the expression of VEGF transcripts in different treatment groups (BC; NC; and 1, 10 and 100 nmol/l ATRA). GAPDH was used as an internal control. (B) Enzyme-linked immunosorbent assay analysis was used to determine protein expression levels of VEGF in different treatment groups (BC; NC; and 1, 10 and 100 nmol/l ATRA). Values are presented as the mean ± standard deviation. * P

    Techniques Used: Expressing, Incubation, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Standard Deviation

    12) Product Images from "Human Platelet Lysates‐Based Hydrogels: A Novel Personalized 3D Platform for Spheroid Invasion Assessment, Human Platelet Lysates‐Based Hydrogels: A Novel Personalized 3D Platform for Spheroid Invasion Assessment"

    Article Title: Human Platelet Lysates‐Based Hydrogels: A Novel Personalized 3D Platform for Spheroid Invasion Assessment, Human Platelet Lysates‐Based Hydrogels: A Novel Personalized 3D Platform for Spheroid Invasion Assessment

    Journal: Advanced Science

    doi: 10.1002/advs.201902398

    Protein and growth factor release from PLMA hydrogels. A) Total protein quantification and B,C) ELISA quantification of TGF‐β1 and VEGF‐A release fro m PLMA hydrogels at 10, 15, and 20% (w/v). Data are presented as mean ± SD ( n ≥ 3).
    Figure Legend Snippet: Protein and growth factor release from PLMA hydrogels. A) Total protein quantification and B,C) ELISA quantification of TGF‐β1 and VEGF‐A release fro m PLMA hydrogels at 10, 15, and 20% (w/v). Data are presented as mean ± SD ( n ≥ 3).

    Techniques Used: Enzyme-linked Immunosorbent Assay

    13) Product Images from "Microencapsulation of low-passage poorly-differentiated human mucoepidermoid carcinoma cells by alginate microcapsules: in vitro profiling of angiogenesis-related molecules"

    Article Title: Microencapsulation of low-passage poorly-differentiated human mucoepidermoid carcinoma cells by alginate microcapsules: in vitro profiling of angiogenesis-related molecules

    Journal: Cancer Cell International

    doi: 10.1186/s12935-017-0479-6

    The release of the pro-angiogenic inducer VEGF-A from 2D versus 3D cultured cancer cells. Quantitative analysis with the human VEGF-A ELISA kit. Three independent repeats, with each duplicates, were performed. 2D vs. 3D: *** P
    Figure Legend Snippet: The release of the pro-angiogenic inducer VEGF-A from 2D versus 3D cultured cancer cells. Quantitative analysis with the human VEGF-A ELISA kit. Three independent repeats, with each duplicates, were performed. 2D vs. 3D: *** P

    Techniques Used: Cell Culture, Enzyme-linked Immunosorbent Assay

    14) Product Images from "Phosphorylation of STAT3 and ERBB2 mediates hypoxia-induced VEGF release in ARPE-19 cells"

    Article Title: Phosphorylation of STAT3 and ERBB2 mediates hypoxia-induced VEGF release in ARPE-19 cells

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2020.11344

    AG490 suppresses hypoxia-induced VEGF release. ARPE-19 cells were exposed to hypoxia and treated with the indicated concentrations of AG490 for 24 h. VEGF release was measured by ELISA. ***P
    Figure Legend Snippet: AG490 suppresses hypoxia-induced VEGF release. ARPE-19 cells were exposed to hypoxia and treated with the indicated concentrations of AG490 for 24 h. VEGF release was measured by ELISA. ***P

    Techniques Used: Enzyme-linked Immunosorbent Assay

    15) Product Images from "Cytoprotective Effects of Natural Highly Bio-Available Vegetable Derivatives on Human-Derived Retinal Cells"

    Article Title: Cytoprotective Effects of Natural Highly Bio-Available Vegetable Derivatives on Human-Derived Retinal Cells

    Journal: Nutrients

    doi: 10.3390/nu12030879

    Effect of curcumin, lutein, and resveratrol on vascular endothelial growth factor (VEGF) secretion. VEGF amounts were quantified by ELISA in the cell supernatants after a 24 h incubation with natural agents, alone or combined. Results are expressed as mean percentage ± SEM. The number of tests for each condition is indicated in each bar. Each n corresponds to a triplicate. Statistical significance (*) compared to “0”.
    Figure Legend Snippet: Effect of curcumin, lutein, and resveratrol on vascular endothelial growth factor (VEGF) secretion. VEGF amounts were quantified by ELISA in the cell supernatants after a 24 h incubation with natural agents, alone or combined. Results are expressed as mean percentage ± SEM. The number of tests for each condition is indicated in each bar. Each n corresponds to a triplicate. Statistical significance (*) compared to “0”.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Incubation

    Effect of curcumin and lutein on the prevention of hypoxia-induced VEGF secretion. ARPE-19 cells were treated or untreated with curcumin + lutein for 24 h before induction of hypoxia with CoCl 2 at 200 µM for 4 h. ( a ). VEGF quantification by ELISA in supernatants. ( b ). Cell viability and ( c ). caspase 3/7 activity ( n = 4, triplicates per group). Results are expressed as mean % ± SEM. Statistical significance (*) compared to No-CoCl 2 -No-pretreatment or (†) compared to CoCl 2 -No-Pretreatment.
    Figure Legend Snippet: Effect of curcumin and lutein on the prevention of hypoxia-induced VEGF secretion. ARPE-19 cells were treated or untreated with curcumin + lutein for 24 h before induction of hypoxia with CoCl 2 at 200 µM for 4 h. ( a ). VEGF quantification by ELISA in supernatants. ( b ). Cell viability and ( c ). caspase 3/7 activity ( n = 4, triplicates per group). Results are expressed as mean % ± SEM. Statistical significance (*) compared to No-CoCl 2 -No-pretreatment or (†) compared to CoCl 2 -No-Pretreatment.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Activity Assay

    16) Product Images from "Human Motor Neuron Progenitor Transplantation Leads to Endogenous Neuronal Sparing in 3 Models of Motor Neuron Loss"

    Article Title: Human Motor Neuron Progenitor Transplantation Leads to Endogenous Neuronal Sparing in 3 Models of Motor Neuron Loss

    Journal: Stem Cells International

    doi: 10.4061/2011/207230

    hMNPs secrete physiologically active growth factors in vivo . ELISA data of NGF (a), NT-3 (b), VEGF (c), and NT-4 (d) protein levels in the thoracic spinal cord transplanted region of SCI rats. Data is expressed as mean ± standard error. * P
    Figure Legend Snippet: hMNPs secrete physiologically active growth factors in vivo . ELISA data of NGF (a), NT-3 (b), VEGF (c), and NT-4 (d) protein levels in the thoracic spinal cord transplanted region of SCI rats. Data is expressed as mean ± standard error. * P

    Techniques Used: In Vivo, Enzyme-linked Immunosorbent Assay

    17) Product Images from "Viburnum opulus L. Juice Phenolic Compounds Influence Osteogenic Differentiation in Human Osteosarcoma Saos-2 Cells"

    Article Title: Viburnum opulus L. Juice Phenolic Compounds Influence Osteogenic Differentiation in Human Osteosarcoma Saos-2 Cells

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21144909

    The influence V. opulus FJ and PJ at IC 0 concentration on cellular metabolic activity, determined with Presto Blue assay ( A ) and VEGF secretion quantified with ELISA ( B ) by HUVEC cells; control cells were not exposed to any compound; values are means ± SEM, n ≥ 3. Morphology of HUVEC cells treated with V. opulus for 48 h ( C ); randomly chosen fields were photographed at ×200 magnification.
    Figure Legend Snippet: The influence V. opulus FJ and PJ at IC 0 concentration on cellular metabolic activity, determined with Presto Blue assay ( A ) and VEGF secretion quantified with ELISA ( B ) by HUVEC cells; control cells were not exposed to any compound; values are means ± SEM, n ≥ 3. Morphology of HUVEC cells treated with V. opulus for 48 h ( C ); randomly chosen fields were photographed at ×200 magnification.

    Techniques Used: Concentration Assay, Activity Assay, Enzyme-linked Immunosorbent Assay

    18) Product Images from "Rational design of therapeutic mAbs against aggregation through protein engineering and incorporation of glycosylation motifs applied to bevacizumab"

    Article Title: Rational design of therapeutic mAbs against aggregation through protein engineering and incorporation of glycosylation motifs applied to bevacizumab

    Journal: mAbs

    doi: 10.1080/19420862.2015.1112477

    Competitive ELISA of WT bevacizumab and single variants for VEGF-A. Bevacizumab and engineered proteins binding to VEGF were measured using a VEGF Human ELISA Kit in which the VEGF protein (1500 pg/mL) was pre-incubated and shacked for 60 min at 37°C with bevacizumab or the variants at various concentrations (0.5 pM to 1 μM). The pre-mixes were then used in the ELISA, which allows the quantification of antibodies bound to the VEGF by reading absorbance at 450 nm. The more bevacizumab (or variants) binds to VEGF, the less VEGF can bind to the anti-VEGF IgG1 adhered onto the ELISA plate, and the lower the signal at 450 nm. Each assay was run at least in duplicate. The WT and variants bound to the target VEGF with the same affinity, as previously reported for bevacizumab. ( A ) Affinity of WT bevacizumab and the high SAP single point mutation variants for VEGF-A. ( B ) Affinity of WT bevacizumab and hyperglycosylated variants for VEGF-A.
    Figure Legend Snippet: Competitive ELISA of WT bevacizumab and single variants for VEGF-A. Bevacizumab and engineered proteins binding to VEGF were measured using a VEGF Human ELISA Kit in which the VEGF protein (1500 pg/mL) was pre-incubated and shacked for 60 min at 37°C with bevacizumab or the variants at various concentrations (0.5 pM to 1 μM). The pre-mixes were then used in the ELISA, which allows the quantification of antibodies bound to the VEGF by reading absorbance at 450 nm. The more bevacizumab (or variants) binds to VEGF, the less VEGF can bind to the anti-VEGF IgG1 adhered onto the ELISA plate, and the lower the signal at 450 nm. Each assay was run at least in duplicate. The WT and variants bound to the target VEGF with the same affinity, as previously reported for bevacizumab. ( A ) Affinity of WT bevacizumab and the high SAP single point mutation variants for VEGF-A. ( B ) Affinity of WT bevacizumab and hyperglycosylated variants for VEGF-A.

    Techniques Used: Competitive ELISA, Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation, Mutagenesis

    19) Product Images from "An Engineered Optogenetic Switch for Spatiotemporal Control of Gene Expression, Cell Differentiation, and Tissue Morphogenesis"

    Article Title: An Engineered Optogenetic Switch for Spatiotemporal Control of Gene Expression, Cell Differentiation, and Tissue Morphogenesis

    Journal: ACS synthetic biology

    doi: 10.1021/acssynbio.7b00147

    Validation of the Flox VPA lentiviral vector. (A) Cells expressing Flox VPA and the light-inducible CRE constructs perform as a permanent light-inducible “on” switch. HEK293T cells were transduced with Flox VPA and transfected with an empty plasmid (Flox Only), a plasmid that constitutively expresses Cre recombinase (CMV-Cre), CRY2-Cre N and CIB1 N -Cre C (CIB1 N Light and CIB1 N Dark), or CRY2-Cre N and CIB1 FL -Cre C (CIB1 FL Light and CIB1 FL Dark). After 24 hours, cells were either illuminated for three days and then incubated in the dark for 9 days (Light) or incubated in the dark for the entire experiment (Dark). Unmodified cells that were transfected only with an empty plasmid served as a negative control (Mock). VEGF (top panel) and ANG1 (bottom panel) expression was determined by ELISA. (B) qRT-PCR was used to detect the levels of recombination of Flox VPA using primers specific for the recombined mRNA. Samples were harvested after two (blue) and seven (red) days of illumination. Conditions not connected by the same letter are significantly different as determined by Tukey’s HSD test. BD, DE, CD, BC: p
    Figure Legend Snippet: Validation of the Flox VPA lentiviral vector. (A) Cells expressing Flox VPA and the light-inducible CRE constructs perform as a permanent light-inducible “on” switch. HEK293T cells were transduced with Flox VPA and transfected with an empty plasmid (Flox Only), a plasmid that constitutively expresses Cre recombinase (CMV-Cre), CRY2-Cre N and CIB1 N -Cre C (CIB1 N Light and CIB1 N Dark), or CRY2-Cre N and CIB1 FL -Cre C (CIB1 FL Light and CIB1 FL Dark). After 24 hours, cells were either illuminated for three days and then incubated in the dark for 9 days (Light) or incubated in the dark for the entire experiment (Dark). Unmodified cells that were transfected only with an empty plasmid served as a negative control (Mock). VEGF (top panel) and ANG1 (bottom panel) expression was determined by ELISA. (B) qRT-PCR was used to detect the levels of recombination of Flox VPA using primers specific for the recombined mRNA. Samples were harvested after two (blue) and seven (red) days of illumination. Conditions not connected by the same letter are significantly different as determined by Tukey’s HSD test. BD, DE, CD, BC: p

    Techniques Used: Plasmid Preparation, Expressing, Construct, Transduction, Transfection, Incubation, Negative Control, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

    20) Product Images from "SP1/TGF-β1/SMAD2 pathway is involved in angiogenesis during osteogenesis"

    Article Title: SP1/TGF-β1/SMAD2 pathway is involved in angiogenesis during osteogenesis

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2020.10965

    SP1 promotes angiogenesis in preosteoblasts through activating the TGF-β1/SMAD2 pathway. (A and B) MC3T3-E1 cells were transfected with p-SP1 overexpression vector and co-treated with SB431542. (A) mRNA and (B) protein expression levels of TGF-β1 were detected by reverse transcription-quantitative PCR and western blotting, respectively. (C) MC3T3-E1 cells were transfected with p-SP1 and co-treated with SB431542 and the protein expression levels of SMAD2 and p-SMAD2 were detected and semi-quantified using western blotting. GAPDH served as the loading control. (D) MC3T3-E1 cells were transfected with p-SP1 and co-treated with SB431542 and the VEGF concentration in supernatants were detected using ELISA. (E-G) MC3T3-E1 cells were transfected with p-SP1 and co-treated with SB431542, and co-cultured with HUVECs. (E) The cell viability of HUVECs under each condition was detected using an MTT assay. (F) Migratory or (G) angiogenic ability of HUVECs under each condition was assessed by the Transwell assay and angiogenesis assay, respectively. Representative micrographs are presented for HUVECs under each condition. Scale bar = 100 µm. Data are presented as the mean ± SD. *P
    Figure Legend Snippet: SP1 promotes angiogenesis in preosteoblasts through activating the TGF-β1/SMAD2 pathway. (A and B) MC3T3-E1 cells were transfected with p-SP1 overexpression vector and co-treated with SB431542. (A) mRNA and (B) protein expression levels of TGF-β1 were detected by reverse transcription-quantitative PCR and western blotting, respectively. (C) MC3T3-E1 cells were transfected with p-SP1 and co-treated with SB431542 and the protein expression levels of SMAD2 and p-SMAD2 were detected and semi-quantified using western blotting. GAPDH served as the loading control. (D) MC3T3-E1 cells were transfected with p-SP1 and co-treated with SB431542 and the VEGF concentration in supernatants were detected using ELISA. (E-G) MC3T3-E1 cells were transfected with p-SP1 and co-treated with SB431542, and co-cultured with HUVECs. (E) The cell viability of HUVECs under each condition was detected using an MTT assay. (F) Migratory or (G) angiogenic ability of HUVECs under each condition was assessed by the Transwell assay and angiogenesis assay, respectively. Representative micrographs are presented for HUVECs under each condition. Scale bar = 100 µm. Data are presented as the mean ± SD. *P

    Techniques Used: Transfection, Over Expression, Plasmid Preparation, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, MTT Assay, Transwell Assay, Angiogenesis Assay

    SP1 promotes angiogenesis in preosteoblasts. (A and B) MC3T3-E1 cells were transfected with p-SP1 or p-NC overexpression vectors and the (A) mRNA and (B) protein expression levels of SP1 and VEGF were analyzed using reverse transcription-quantitative PCR and western blotting, respectively. (C) MC3T3-E1 cells were transfected with p-SP1 or p-NC. Supernatants were collected and the protein expression levels of VEGF in the supernatants were detected using ELISA. (D-F) MC3T3-E1 cells were transfected with p-SP1 or p-NC and co-cultured with HUVECs. (D) The viability of HUVECs was detected using the MTT assay. (E) The migratory ability of HUVECs was detected using the Transwell assay. Representative micrographs are provided of HUVECs under each condition. Scale bar=100 µm. (F) The capillary tube formation rate of HUVECs was detected using an angiogenesis assay. Scale bar=100 µm. Representative micrographs are provided of HUVECs under each condition. Data represents one of the three independent experiments replicates with n=3/group for each experimental group of. Data are presented as the mean ± SD. *P
    Figure Legend Snippet: SP1 promotes angiogenesis in preosteoblasts. (A and B) MC3T3-E1 cells were transfected with p-SP1 or p-NC overexpression vectors and the (A) mRNA and (B) protein expression levels of SP1 and VEGF were analyzed using reverse transcription-quantitative PCR and western blotting, respectively. (C) MC3T3-E1 cells were transfected with p-SP1 or p-NC. Supernatants were collected and the protein expression levels of VEGF in the supernatants were detected using ELISA. (D-F) MC3T3-E1 cells were transfected with p-SP1 or p-NC and co-cultured with HUVECs. (D) The viability of HUVECs was detected using the MTT assay. (E) The migratory ability of HUVECs was detected using the Transwell assay. Representative micrographs are provided of HUVECs under each condition. Scale bar=100 µm. (F) The capillary tube formation rate of HUVECs was detected using an angiogenesis assay. Scale bar=100 µm. Representative micrographs are provided of HUVECs under each condition. Data represents one of the three independent experiments replicates with n=3/group for each experimental group of. Data are presented as the mean ± SD. *P

    Techniques Used: Transfection, Over Expression, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Culture, MTT Assay, Transwell Assay, Angiogenesis Assay

    SP1/TGF-β1/SMAD2 pathway promotes preosteoblast angiogenesis through regulating VEGF expression. (A-D) MC3T3-E1 cells were transfected with p-SP1 overexpression vector and treated with VEGF, SB431542, or bevacizumab in different combinations as indicated and co-cultured with HUVECs. (A) The VEGF concentration in the supernatants was detected by ELISA 24 h post-treatment. (B) Cell viability was detected using an MTT assay. (C) Migratory ability of these cells was detected using a Transwell assay. Representative micrographs of HUVECs under each treatment condition are provided. Scale bar=100 µm. (D) The ability to undergo angiogenesis was determined using an angiogenesis assay. Representative micrographs of HUVECs under each treatment condition are provided. Data are presented as the mean ± SD. *P
    Figure Legend Snippet: SP1/TGF-β1/SMAD2 pathway promotes preosteoblast angiogenesis through regulating VEGF expression. (A-D) MC3T3-E1 cells were transfected with p-SP1 overexpression vector and treated with VEGF, SB431542, or bevacizumab in different combinations as indicated and co-cultured with HUVECs. (A) The VEGF concentration in the supernatants was detected by ELISA 24 h post-treatment. (B) Cell viability was detected using an MTT assay. (C) Migratory ability of these cells was detected using a Transwell assay. Representative micrographs of HUVECs under each treatment condition are provided. Scale bar=100 µm. (D) The ability to undergo angiogenesis was determined using an angiogenesis assay. Representative micrographs of HUVECs under each treatment condition are provided. Data are presented as the mean ± SD. *P

    Techniques Used: Expressing, Transfection, Over Expression, Plasmid Preparation, Cell Culture, Concentration Assay, Enzyme-linked Immunosorbent Assay, MTT Assay, Transwell Assay, Angiogenesis Assay

    TGF-β1 promotes angiogenesis in preosteoblasts through promoting SMAD2 phosphorylation. (A) MC3T3-E1 cells were treated with TGF-β1 or TGF-β1 + SB431542, and the protein expression levels of SMAD2 and p-SMAD2 were detected and semi-quantified using western blotting. GAPDH served as a loading control. Statistical analyses are given in the bar charts. (B) MC3T3-E1 cells were treated as described in (A) and protein expression levels of VEGF in the supernatants were detected by ELISA. (C-E) MC3T3-E1 cells were treated with TGF-β1 or TGF-β1 + SB431542 and subsequently co-cultured with HUVECs. (C) Cell viability of these HUVECs was detected using the MTT assay. (D) The migratory ability of these HUVECs were detected using a Transwell assay. Representative micrographs of HUVECs under each condition are presented. Scale bar=100 µm. (E) The ability of the HUVECs under each condition to undergo angiogenesis were analyzed using an angiogenesis assay. Representative micrographs of HUVECs under each condition are presented. Scale bar=100 µm. Data are presented as the mean ± SD. *P
    Figure Legend Snippet: TGF-β1 promotes angiogenesis in preosteoblasts through promoting SMAD2 phosphorylation. (A) MC3T3-E1 cells were treated with TGF-β1 or TGF-β1 + SB431542, and the protein expression levels of SMAD2 and p-SMAD2 were detected and semi-quantified using western blotting. GAPDH served as a loading control. Statistical analyses are given in the bar charts. (B) MC3T3-E1 cells were treated as described in (A) and protein expression levels of VEGF in the supernatants were detected by ELISA. (C-E) MC3T3-E1 cells were treated with TGF-β1 or TGF-β1 + SB431542 and subsequently co-cultured with HUVECs. (C) Cell viability of these HUVECs was detected using the MTT assay. (D) The migratory ability of these HUVECs were detected using a Transwell assay. Representative micrographs of HUVECs under each condition are presented. Scale bar=100 µm. (E) The ability of the HUVECs under each condition to undergo angiogenesis were analyzed using an angiogenesis assay. Representative micrographs of HUVECs under each condition are presented. Scale bar=100 µm. Data are presented as the mean ± SD. *P

    Techniques Used: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Culture, MTT Assay, Transwell Assay, Angiogenesis Assay

    21) Product Images from "Tetrandrine suppresses lung cancer growth and induces apoptosis, potentially via the VEGF/HIF-1α/ICAM-1 signaling pathway"

    Article Title: Tetrandrine suppresses lung cancer growth and induces apoptosis, potentially via the VEGF/HIF-1α/ICAM-1 signaling pathway

    Journal: Oncology Letters

    doi: 10.3892/ol.2018.8190

    Tetrandrine increases the expression of vascular endothelial growth factor, as determined using an ELISA. *P
    Figure Legend Snippet: Tetrandrine increases the expression of vascular endothelial growth factor, as determined using an ELISA. *P

    Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay

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    Thermo Fisher human vegf a elisa kit
    (A) Cytotoxicity of HOHA-lactone to ARPE-19 cells (25,000 cells/well). Cells were treated with 0–50 µM HOHA-lactone for 2 h and then assayed for cytotoxicity by MTT assay. Cell survival values (%) were presented as means ± S.D. of 8 independent experiments. (B) Effect of HOHA-lactone on VEGF secretion by ARPE-19 cells. ARPE-19 cells grown in a 96-well plate (25,000 cells per well) were treated with serum-free culture media containing varying concentrations (0–25 µM) of HOHA-lactone. After 12 h of incubation, supernatants were collected and secreted VEGF was measured using a human <t>VEGF-A</t> <t>ELISA</t> kit. The data represent the mean ± SD (n = 8
    Human Vegf A Elisa Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 79 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    (A) Cytotoxicity of HOHA-lactone to ARPE-19 cells (25,000 cells/well). Cells were treated with 0–50 µM HOHA-lactone for 2 h and then assayed for cytotoxicity by MTT assay. Cell survival values (%) were presented as means ± S.D. of 8 independent experiments. (B) Effect of HOHA-lactone on VEGF secretion by ARPE-19 cells. ARPE-19 cells grown in a 96-well plate (25,000 cells per well) were treated with serum-free culture media containing varying concentrations (0–25 µM) of HOHA-lactone. After 12 h of incubation, supernatants were collected and secreted VEGF was measured using a human VEGF-A ELISA kit. The data represent the mean ± SD (n = 8

    Journal: Chemical research in toxicology

    Article Title: 4-Hydroxy-7-oxo-5-heptenoic Acid (HOHA) Lactone is a Biologically Active Precursor for the Generation of 2-(ω-Carboxyethyl)pyrrole (CEP) Derivatives of Proteins and Ethanolamine Phospholipids

    doi: 10.1021/acs.chemrestox.5b00001

    Figure Lengend Snippet: (A) Cytotoxicity of HOHA-lactone to ARPE-19 cells (25,000 cells/well). Cells were treated with 0–50 µM HOHA-lactone for 2 h and then assayed for cytotoxicity by MTT assay. Cell survival values (%) were presented as means ± S.D. of 8 independent experiments. (B) Effect of HOHA-lactone on VEGF secretion by ARPE-19 cells. ARPE-19 cells grown in a 96-well plate (25,000 cells per well) were treated with serum-free culture media containing varying concentrations (0–25 µM) of HOHA-lactone. After 12 h of incubation, supernatants were collected and secreted VEGF was measured using a human VEGF-A ELISA kit. The data represent the mean ± SD (n = 8

    Article Snippet: After 12 h, supernatants were collected to estimate secreted VEGF using an enzyme-linked immunosorbent assay (ELISA) according to the manufacturers protocol (Human VEGF-A ELISA kit, Thermo Scientific, Rockford, IL).

    Techniques: MTT Assay, Incubation, Enzyme-linked Immunosorbent Assay

    PERK activation stimulated medulloblastoma cells to produce VEGF-A. (A) Real-time PCR analysis showed that 0.01 nM AP20187 treatment significantly increased the expression of CHOP, GADD34, and VEGF-A in Fv2E-PERK1 cells, but did not affect the expression of VEGFR2 and BIP. (B) ELISA analysis showed that 0.01 nM AP20187 treatment significantly increased the production of VEGF-A in Fv2E-PERK1 cells. (C) Real-time PCR analysis showed that 10 μM Salubrinal treatment significantly increased the expression of CHOP, GADD34, and VEGF-A in Daoy cells, but did not affect the expression of VEGFR2 and BIP. (D) ELISA analysis showed that 10 μM Salubrinal treatment significantly increased the production of VEGF-A in Daoy cells. (E) Real-time PCR analysis showed that 10 μM Salubrinal treatment significantly increased the expression of CHOP, GADD34, and VEGF-A in UW228 cells, but did not affect the expression of VEGFR2 and BIP. (F) ELISA analysis showed that 10 μM Salubrinal treatment significantly increased the production of VEGF-A in UW228 cells. The experiments were repeated at least three times. Error bars represent SD, * P

    Journal: PLoS ONE

    Article Title: Pancreatic Endoplasmic Reticulum Kinase Activation Promotes Medulloblastoma Cell Migration and Invasion through Induction of Vascular Endothelial Growth Factor A

    doi: 10.1371/journal.pone.0120252

    Figure Lengend Snippet: PERK activation stimulated medulloblastoma cells to produce VEGF-A. (A) Real-time PCR analysis showed that 0.01 nM AP20187 treatment significantly increased the expression of CHOP, GADD34, and VEGF-A in Fv2E-PERK1 cells, but did not affect the expression of VEGFR2 and BIP. (B) ELISA analysis showed that 0.01 nM AP20187 treatment significantly increased the production of VEGF-A in Fv2E-PERK1 cells. (C) Real-time PCR analysis showed that 10 μM Salubrinal treatment significantly increased the expression of CHOP, GADD34, and VEGF-A in Daoy cells, but did not affect the expression of VEGFR2 and BIP. (D) ELISA analysis showed that 10 μM Salubrinal treatment significantly increased the production of VEGF-A in Daoy cells. (E) Real-time PCR analysis showed that 10 μM Salubrinal treatment significantly increased the expression of CHOP, GADD34, and VEGF-A in UW228 cells, but did not affect the expression of VEGFR2 and BIP. (F) ELISA analysis showed that 10 μM Salubrinal treatment significantly increased the production of VEGF-A in UW228 cells. The experiments were repeated at least three times. Error bars represent SD, * P

    Article Snippet: After 24 h, the cells were raised with PBS, and then incubated in DMEM with 0.5% FBS and various treatments for 16 h. We quantified VEGF-A in the culture supernatants of the wells using human VEGF-A ELISA kit (Thermo Scientific) according to the manufacturer’s instructions.

    Techniques: Activation Assay, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay

    Sandwich ELISA assay measuring concentration of VEGF in (a) ARPE-19 and (b) D407 cell culture supernatant post exposure to hypoxia (12 h) in the absence or presence of ritonavir (5, 10, and 20 μ M). Values are represented as mean±SD. A P -value of

    Journal: Eye

    Article Title: Ritonavir inhibits HIF-1α-mediated VEGF expression in retinal pigment epithelial cells in vitro

    doi: 10.1038/eye.2013.240

    Figure Lengend Snippet: Sandwich ELISA assay measuring concentration of VEGF in (a) ARPE-19 and (b) D407 cell culture supernatant post exposure to hypoxia (12 h) in the absence or presence of ritonavir (5, 10, and 20 μ M). Values are represented as mean±SD. A P -value of

    Article Snippet: BCA protein assay reagent and human VEGF ELISA kit were purchased from Thermo Fisher Scientific Inc. (Rockford, IL, USA).

    Techniques: Sandwich ELISA, Concentration Assay, Cell Culture

    ( A ) LMB-100 treatment reduces levels of CCSGFs in conditioned medium, while paclitaxel increases it. For experiments in ( B – D ), the indicated pancreatic cancer cell lines were treated with LMB-100 for 24 h as per schema. Medium was replaced to stop treatment then collected 24 h later. ( B ) Conditioned medium from treated cells were assayed for VEGF using ELISA assay. ( C ) Conditioned medium from KLM1 cells was assayed for multiple CCSGFs by human analyte Luminex assay. ( D ) Conditioned medium from Panc02-chiMSLN cells was assayed for multiple CCSGFs by murine analyte Luminex assay. ( E ) Viable cells from triplicate wells were treated with paclitaxel for 24 h as shown in schema and then counted to assess viability. ( F ) Conditioned medium from KLM1 cells treated with paclitaxel were assayed for multiple CCSGFs by human analyte Luminex assay.

    Journal: Toxins

    Article Title: Protein Synthesis Inhibition Activity of Mesothelin Targeting Immunotoxin LMB-100 Decreases Concentrations of Oncogenic Signaling Molecules and Secreted Growth Factors

    doi: 10.3390/toxins10110447

    Figure Lengend Snippet: ( A ) LMB-100 treatment reduces levels of CCSGFs in conditioned medium, while paclitaxel increases it. For experiments in ( B – D ), the indicated pancreatic cancer cell lines were treated with LMB-100 for 24 h as per schema. Medium was replaced to stop treatment then collected 24 h later. ( B ) Conditioned medium from treated cells were assayed for VEGF using ELISA assay. ( C ) Conditioned medium from KLM1 cells was assayed for multiple CCSGFs by human analyte Luminex assay. ( D ) Conditioned medium from Panc02-chiMSLN cells was assayed for multiple CCSGFs by murine analyte Luminex assay. ( E ) Viable cells from triplicate wells were treated with paclitaxel for 24 h as shown in schema and then counted to assess viability. ( F ) Conditioned medium from KLM1 cells treated with paclitaxel were assayed for multiple CCSGFs by human analyte Luminex assay.

    Article Snippet: Human VEGF ELISA kit was purchased from ThermoFisher.

    Techniques: Enzyme-linked Immunosorbent Assay, Luminex