Structured Review

R&D Systems human free bdnf quantikine elisa kit
Retinal GFP and <t>BDNF</t> expression after intravitreal injection of tm-scAAV2-GFP and tm-scAAV2-BDNF. A : Fluorescence microscopic images of cross sections treated with tm-scAAV2-GFP and NMDA. Green: GFP, Blue: DAPI. Scale bar represents 50 μm. B : Relative RNA expression levels of BDNF analyzed by the comparative threshold cycle method. GAPDH was used to normalize gene expressions (n = 6 in each group, *p<0.05 for a two-tailed Mann–Whitney U test). C : Protein expression levels of BDNF after intravitreal injection of tm-scAAV2-BDNF, as analyzed by <t>ELISA</t> (n = 4 in each group, *p<0.05 for a two-tailed Mann–Whitney U test).
Human Free Bdnf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human free bdnf quantikine elisa kit - by Bioz Stars, 2023-01
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1) Product Images from "Tyrosine triple mutated AAV2-BDNF gene therapy in an inner retinal injury model induced by intravitreal injection of N –methyl-D-aspartate (NMDA)"

Article Title: Tyrosine triple mutated AAV2-BDNF gene therapy in an inner retinal injury model induced by intravitreal injection of N –methyl-D-aspartate (NMDA)

Journal: Molecular Vision

doi:

Retinal GFP and BDNF expression after intravitreal injection of tm-scAAV2-GFP and tm-scAAV2-BDNF. A : Fluorescence microscopic images of cross sections treated with tm-scAAV2-GFP and NMDA. Green: GFP, Blue: DAPI. Scale bar represents 50 μm. B : Relative RNA expression levels of BDNF analyzed by the comparative threshold cycle method. GAPDH was used to normalize gene expressions (n = 6 in each group, *p<0.05 for a two-tailed Mann–Whitney U test). C : Protein expression levels of BDNF after intravitreal injection of tm-scAAV2-BDNF, as analyzed by ELISA (n = 4 in each group, *p<0.05 for a two-tailed Mann–Whitney U test).
Figure Legend Snippet: Retinal GFP and BDNF expression after intravitreal injection of tm-scAAV2-GFP and tm-scAAV2-BDNF. A : Fluorescence microscopic images of cross sections treated with tm-scAAV2-GFP and NMDA. Green: GFP, Blue: DAPI. Scale bar represents 50 μm. B : Relative RNA expression levels of BDNF analyzed by the comparative threshold cycle method. GAPDH was used to normalize gene expressions (n = 6 in each group, *p<0.05 for a two-tailed Mann–Whitney U test). C : Protein expression levels of BDNF after intravitreal injection of tm-scAAV2-BDNF, as analyzed by ELISA (n = 4 in each group, *p<0.05 for a two-tailed Mann–Whitney U test).

Techniques Used: Expressing, Injection, Fluorescence, RNA Expression, Two Tailed Test, MANN-WHITNEY, Enzyme-linked Immunosorbent Assay


Structured Review

R&D Systems quantikine human free bdnf elisa kit
Quantikine Human Free Bdnf Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/quantikine human free bdnf elisa kit/product/R&D Systems
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
quantikine human free bdnf elisa kit - by Bioz Stars, 2023-01
86/100 stars

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Structured Review

R&D Systems human free bdnf quantikine elisa kit
Transfection of ARPE-19 cells using different ratios of SB100X transposase plasmid and <t>BDNF</t> transposon plasmid. In each experiment ( n = 6), 1 × 10 5 cells were transfected with an increasing ratio (1:1–1:24) of SB100X transposase to BDNF transposon plasmid at a total concentration of 0.5 µg, plus one control without the addition of plasmid DNA (Co). Cultures were terminated 7.33 ± 0.52 days after transfection. Cells were used for the isolation of total RNA. Medium used for western blot and <t>ELISA</t> was added to the cells in a defined volume 24 h before culture termination. ( a ) BDNF secretion was analyzed by western blots of the culture medium. Loading of equal amounts of supernatant was proven by Stain-Free imaging technology. Each western blot signal was referenced to the total protein content of the sample and normalized to the mean value of the respective experiment. BDNF signals were more evident in transfected cultures than in Co cultures (ns: not significant, * p < 0.05, ** p < 0.01, Brown-Forsythe and Welch ANOVA tests with Dunnett’s T3 multiple comparisons test). ( b ) BDNF gene expression was analyzed by qPCR. Total (endogenous + recombinant) BDNF expression in transfected cultures was related to the expression in Co cultures, which was set to 1 (dashed line). An increase in BDNF gene expression was observed in the transfected cultures (ns: not significant, * p < 0.05, ** p < 0.01, *** p = 0.0008, one sample t test). ( c ) BDNF secretion was quantified by ELISA using precisely defined cell culture supernatants. Values of the transfected cultures were higher than those of the Co cultures (ns: not significant, * p < 0.05, ** p = 0.004, Brown-Forsythe and Welch ANOVA tests with Dunnett’s T3 multiple comparisons test). Data are presented as box and whisker plots (whiskers: minimum to maximum, mean values indicated by +). Outliers were evaluated using the ROUT method (Q = 1%, indicated by red open circles).
Human Free Bdnf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human free bdnf quantikine elisa kit/product/R&D Systems
Average 94 stars, based on 1 article reviews
Price from $9.99 to $1999.99
human free bdnf quantikine elisa kit - by Bioz Stars, 2023-01
94/100 stars

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1) Product Images from "Molecular and Functional Characterization of BDNF-Overexpressing Human Retinal Pigment Epithelial Cells Established by Sleeping Beauty Transposon-Mediated Gene Transfer"

Article Title: Molecular and Functional Characterization of BDNF-Overexpressing Human Retinal Pigment Epithelial Cells Established by Sleeping Beauty Transposon-Mediated Gene Transfer

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms232112982

Transfection of ARPE-19 cells using different ratios of SB100X transposase plasmid and BDNF transposon plasmid. In each experiment ( n = 6), 1 × 10 5 cells were transfected with an increasing ratio (1:1–1:24) of SB100X transposase to BDNF transposon plasmid at a total concentration of 0.5 µg, plus one control without the addition of plasmid DNA (Co). Cultures were terminated 7.33 ± 0.52 days after transfection. Cells were used for the isolation of total RNA. Medium used for western blot and ELISA was added to the cells in a defined volume 24 h before culture termination. ( a ) BDNF secretion was analyzed by western blots of the culture medium. Loading of equal amounts of supernatant was proven by Stain-Free imaging technology. Each western blot signal was referenced to the total protein content of the sample and normalized to the mean value of the respective experiment. BDNF signals were more evident in transfected cultures than in Co cultures (ns: not significant, * p < 0.05, ** p < 0.01, Brown-Forsythe and Welch ANOVA tests with Dunnett’s T3 multiple comparisons test). ( b ) BDNF gene expression was analyzed by qPCR. Total (endogenous + recombinant) BDNF expression in transfected cultures was related to the expression in Co cultures, which was set to 1 (dashed line). An increase in BDNF gene expression was observed in the transfected cultures (ns: not significant, * p < 0.05, ** p < 0.01, *** p = 0.0008, one sample t test). ( c ) BDNF secretion was quantified by ELISA using precisely defined cell culture supernatants. Values of the transfected cultures were higher than those of the Co cultures (ns: not significant, * p < 0.05, ** p = 0.004, Brown-Forsythe and Welch ANOVA tests with Dunnett’s T3 multiple comparisons test). Data are presented as box and whisker plots (whiskers: minimum to maximum, mean values indicated by +). Outliers were evaluated using the ROUT method (Q = 1%, indicated by red open circles).
Figure Legend Snippet: Transfection of ARPE-19 cells using different ratios of SB100X transposase plasmid and BDNF transposon plasmid. In each experiment ( n = 6), 1 × 10 5 cells were transfected with an increasing ratio (1:1–1:24) of SB100X transposase to BDNF transposon plasmid at a total concentration of 0.5 µg, plus one control without the addition of plasmid DNA (Co). Cultures were terminated 7.33 ± 0.52 days after transfection. Cells were used for the isolation of total RNA. Medium used for western blot and ELISA was added to the cells in a defined volume 24 h before culture termination. ( a ) BDNF secretion was analyzed by western blots of the culture medium. Loading of equal amounts of supernatant was proven by Stain-Free imaging technology. Each western blot signal was referenced to the total protein content of the sample and normalized to the mean value of the respective experiment. BDNF signals were more evident in transfected cultures than in Co cultures (ns: not significant, * p < 0.05, ** p < 0.01, Brown-Forsythe and Welch ANOVA tests with Dunnett’s T3 multiple comparisons test). ( b ) BDNF gene expression was analyzed by qPCR. Total (endogenous + recombinant) BDNF expression in transfected cultures was related to the expression in Co cultures, which was set to 1 (dashed line). An increase in BDNF gene expression was observed in the transfected cultures (ns: not significant, * p < 0.05, ** p < 0.01, *** p = 0.0008, one sample t test). ( c ) BDNF secretion was quantified by ELISA using precisely defined cell culture supernatants. Values of the transfected cultures were higher than those of the Co cultures (ns: not significant, * p < 0.05, ** p = 0.004, Brown-Forsythe and Welch ANOVA tests with Dunnett’s T3 multiple comparisons test). Data are presented as box and whisker plots (whiskers: minimum to maximum, mean values indicated by +). Outliers were evaluated using the ROUT method (Q = 1%, indicated by red open circles).

Techniques Used: Transfection, Plasmid Preparation, Concentration Assay, Isolation, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Imaging, Expressing, Recombinant, Cell Culture, Whisker Assay

Transposition efficiency in ARPE-19 cells transfected with a defined ratio of SB100X transposase to BDNF transposon plasmid. In each experiment ( n = 7), four transfections were performed using 5 × 10 4 cells and a mixture of 0.03 µg SB100X transposase and 0.47 µg BDNF transposon plasmid (BDNF + ), plus two controls without the addition of plasmid DNA (Co). Cultures were terminated 21.0 ± 0.58 days after transfection. Cells were used for isolation of total RNA and genomic DNA, respectively. Medium used for western blot and ELISA was added to the cells in a defined volume 24 h before culture termination. ( a ) BDNF secretion was analyzed by western blots of culture medium using Stain-Free Gels. Each western blot signal was referenced to the total protein content of the sample and normalized to the mean value of the respective experiment. Signals in BDNF + cultures were increased compared to Co cultures (ns: not significant, summarized data: **** p < 0.0001, unpaired t test with Welch’s correction; single experiments: * p = 0.0127, ** p < 0.01, 2way ANOVA with Šídák’s multiple comparisons test). ( b ) BDNF gene expression was analyzed by qPCR. Endogenous and total BDNF expression in BDNF + cultures were related to the respective expression in Co cultures, which were set to 1 (dashed line). An increase in total BDNF expression was observed in all BDNF + cultures (ns: not significant, summarized data: **** p < 0.0001, unpaired t test with Welch’s correction; single experiments: *** p = 0.0009, **** p < 0.0001, 2way ANOVA with Šídák’s multiple comparisons test). ( c ) BDNF secretion was quantified by ELISA using precisely defined cell culture supernatants. Values of the BDNF + cultures were higher than those of the Co cultures (ns: not significant, summarized data: **** p < 0.0001, unpaired t test with Welch’s correction; single experiments: *** p = 0.0002, **** p < 0.0001, 2way ANOVA with Šídák’s multiple comparisons test). ( d ) BDNF transposon copy numbers were determined by ddPCR. Average copy numbers per diploid genome in BDNF + cultures were significantly higher compared to the Co cultures (ns: not significant, summarized data: **** p < 0.0001, unpaired t test with Welch’s correction; single experiments: *** p = 0.0007, **** p < 0.0001, 2way ANOVA with Šídák’s multiple comparisons test). Data are presented as box and whisker plots (whiskers: minimum to maximum, mean values indicated by +) and as individual values per experiment (line at mean).
Figure Legend Snippet: Transposition efficiency in ARPE-19 cells transfected with a defined ratio of SB100X transposase to BDNF transposon plasmid. In each experiment ( n = 7), four transfections were performed using 5 × 10 4 cells and a mixture of 0.03 µg SB100X transposase and 0.47 µg BDNF transposon plasmid (BDNF + ), plus two controls without the addition of plasmid DNA (Co). Cultures were terminated 21.0 ± 0.58 days after transfection. Cells were used for isolation of total RNA and genomic DNA, respectively. Medium used for western blot and ELISA was added to the cells in a defined volume 24 h before culture termination. ( a ) BDNF secretion was analyzed by western blots of culture medium using Stain-Free Gels. Each western blot signal was referenced to the total protein content of the sample and normalized to the mean value of the respective experiment. Signals in BDNF + cultures were increased compared to Co cultures (ns: not significant, summarized data: **** p < 0.0001, unpaired t test with Welch’s correction; single experiments: * p = 0.0127, ** p < 0.01, 2way ANOVA with Šídák’s multiple comparisons test). ( b ) BDNF gene expression was analyzed by qPCR. Endogenous and total BDNF expression in BDNF + cultures were related to the respective expression in Co cultures, which were set to 1 (dashed line). An increase in total BDNF expression was observed in all BDNF + cultures (ns: not significant, summarized data: **** p < 0.0001, unpaired t test with Welch’s correction; single experiments: *** p = 0.0009, **** p < 0.0001, 2way ANOVA with Šídák’s multiple comparisons test). ( c ) BDNF secretion was quantified by ELISA using precisely defined cell culture supernatants. Values of the BDNF + cultures were higher than those of the Co cultures (ns: not significant, summarized data: **** p < 0.0001, unpaired t test with Welch’s correction; single experiments: *** p = 0.0002, **** p < 0.0001, 2way ANOVA with Šídák’s multiple comparisons test). ( d ) BDNF transposon copy numbers were determined by ddPCR. Average copy numbers per diploid genome in BDNF + cultures were significantly higher compared to the Co cultures (ns: not significant, summarized data: **** p < 0.0001, unpaired t test with Welch’s correction; single experiments: *** p = 0.0007, **** p < 0.0001, 2way ANOVA with Šídák’s multiple comparisons test). Data are presented as box and whisker plots (whiskers: minimum to maximum, mean values indicated by +) and as individual values per experiment (line at mean).

Techniques Used: Transfection, Plasmid Preparation, Isolation, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Expressing, Cell Culture, Whisker Assay

Growth behavior of BDNF-transfected ARPE-19 cells without or after H 2 O 2 -induced oxidative stress compared to non-transfected control cells. In each experiment ( n = 3 for cultures without H 2 O 2 exposure, n = 4 for cultures with H 2 O 2 exposure), non-treated cells (Co − ), cells exposed to the electric field without plasmid DNA (Co + ), and BDNF-transfected cells (BDNF + , transfected with 0.03 µg SB100X transposase and 0.47 µg BDNF transposon plasmid) were seeded at an initial density of 2 × 10 4 cells. ( a ) Without H 2 O 2 exposure, individual cultures were terminated after 3, 5, 7, 10, and 12 days. Proliferation was determined by classifying live cell count and total cell count. All values were normalized to the highest measured value, which was set to 100%. At the end of the experiment, comparison of the number of living cells or total cells showed no significant differences, and within each culture, the differences between live cell count and total cell count were also not significant (Kruskal-Wallis test with Dunn’s multiple comparison test). ( b ) Cell viability was analyzed using a luminescence-based assay. At the end of the experiment, BDNF + cultures showed significantly higher values than the Co + cultures (* p = 0.0219, Kruskal-Wallis test with Dunn’s multiple comparisons test). ( c ) In the case of H 2 O 2 treatment, individual cultures were terminated after 1, 2, 4, and 8 days. Cells were used for counting and for isolation of total RNA. Medium used for ELISA was added to the cells in a defined volume 24 h before culture termination. No significant differences in cell number were observed at the end of the experiment (Kruskal-Wallis test with Dunn’s multiple comparison test). Data are presented as mean ± SD. ( d ) BDNF gene expression was analyzed by qPCR. Total BDNF expression in BDNF + cultures without and with H 2 O 2 exposure was related to the expression in the corresponding Co cultures, which was set to 1. Increased BDNF expression was observed in all transfected cultures (* p < 0.05 for BDNF + w/o H 2 O 2 and with H 2 O 2 at days 1 and 2. BDNF secretion in BDNF + cultures was quantified by ELISA using precisely defined cell culture supernatants. Comparison of BDNF + cultures at each culture termination showed significantly increased BDNF expression and significantly increased BDNF secretion for cultures with H 2 O 2 at day 8 and day 4, respectively (* p < 0.05, Mann-Whitney test). ( e ) BAX and BCL2 gene expression were analyzed by qPCR. The BAX / BCL2 expression ratios in BDNF + cultures without and with H 2 O 2 exposure were related to the expression ratios in the corresponding Co cultures, which were set to 1. An increased ratio of BAX / BCL2 expression was observed in the transfected cultures. Comparison of BDNF + cultures without and with H 2 O 2 exposure at each termination date showed no significant differences (Mann-Whitney test). Data are presented as box and whisker plots (whiskers: minimum to maximum, mean values indicated by +).
Figure Legend Snippet: Growth behavior of BDNF-transfected ARPE-19 cells without or after H 2 O 2 -induced oxidative stress compared to non-transfected control cells. In each experiment ( n = 3 for cultures without H 2 O 2 exposure, n = 4 for cultures with H 2 O 2 exposure), non-treated cells (Co − ), cells exposed to the electric field without plasmid DNA (Co + ), and BDNF-transfected cells (BDNF + , transfected with 0.03 µg SB100X transposase and 0.47 µg BDNF transposon plasmid) were seeded at an initial density of 2 × 10 4 cells. ( a ) Without H 2 O 2 exposure, individual cultures were terminated after 3, 5, 7, 10, and 12 days. Proliferation was determined by classifying live cell count and total cell count. All values were normalized to the highest measured value, which was set to 100%. At the end of the experiment, comparison of the number of living cells or total cells showed no significant differences, and within each culture, the differences between live cell count and total cell count were also not significant (Kruskal-Wallis test with Dunn’s multiple comparison test). ( b ) Cell viability was analyzed using a luminescence-based assay. At the end of the experiment, BDNF + cultures showed significantly higher values than the Co + cultures (* p = 0.0219, Kruskal-Wallis test with Dunn’s multiple comparisons test). ( c ) In the case of H 2 O 2 treatment, individual cultures were terminated after 1, 2, 4, and 8 days. Cells were used for counting and for isolation of total RNA. Medium used for ELISA was added to the cells in a defined volume 24 h before culture termination. No significant differences in cell number were observed at the end of the experiment (Kruskal-Wallis test with Dunn’s multiple comparison test). Data are presented as mean ± SD. ( d ) BDNF gene expression was analyzed by qPCR. Total BDNF expression in BDNF + cultures without and with H 2 O 2 exposure was related to the expression in the corresponding Co cultures, which was set to 1. Increased BDNF expression was observed in all transfected cultures (* p < 0.05 for BDNF + w/o H 2 O 2 and with H 2 O 2 at days 1 and 2. BDNF secretion in BDNF + cultures was quantified by ELISA using precisely defined cell culture supernatants. Comparison of BDNF + cultures at each culture termination showed significantly increased BDNF expression and significantly increased BDNF secretion for cultures with H 2 O 2 at day 8 and day 4, respectively (* p < 0.05, Mann-Whitney test). ( e ) BAX and BCL2 gene expression were analyzed by qPCR. The BAX / BCL2 expression ratios in BDNF + cultures without and with H 2 O 2 exposure were related to the expression ratios in the corresponding Co cultures, which were set to 1. An increased ratio of BAX / BCL2 expression was observed in the transfected cultures. Comparison of BDNF + cultures without and with H 2 O 2 exposure at each termination date showed no significant differences (Mann-Whitney test). Data are presented as box and whisker plots (whiskers: minimum to maximum, mean values indicated by +).

Techniques Used: Transfection, Plasmid Preparation, Cell Counting, Luminescence Assay, Isolation, Enzyme-linked Immunosorbent Assay, Expressing, Cell Culture, MANN-WHITNEY, Whisker Assay

Transfection of primary hRPE cells using different ratios of SB100X transposase plasmid and BDNF transposon plasmid. In each experiment ( n = 10), 1 × 10 5 primary RPE cells from a total of eight donors (age: 71.9 ± 8.84 years, gender: five males and three females, time postmortem: 27.6 ± 16.3 h, cultivation time before transfection: 67.3 ± 24.0 days) were transfected with an increasing ratio (1:1–1:24) of SB100X transposase to BDNF transposon plasmid at a total concentration of 0.5 µg, plus one control without the addition of plasmid DNA (Co). Cultures were terminated 36.8 ± 10.5 days after transfection. Cells were used for isolation of total RNA. Medium used for ELISA was added to the cells in a defined volume 24 h before culture termination. ( a ) BDNF gene expression was analyzed by qPCR. Total BDNF expression in transfected cultures was related to the expression in Co cultures, which was set to 1 (dashed line). An increase in BDNF expression was observed in the transfected cultures (ns: not significant, * p < 0.05, one sample t test). ( b ) BDNF secretion was quantified by ELISA using precisely defined cell culture supernatants. Values of the transfected cultures were higher than those of the Co cultures, although not significantly (Brown-Forsythe and Welch ANOVA tests with Dunnett’s T3 multiple comparisons test). Data are presented as box and whisker plots (whiskers: minimum to maximum, mean values indicated by +). Outliers were evaluated using the ROUT method (Q = 1%, indicated by red open circles).
Figure Legend Snippet: Transfection of primary hRPE cells using different ratios of SB100X transposase plasmid and BDNF transposon plasmid. In each experiment ( n = 10), 1 × 10 5 primary RPE cells from a total of eight donors (age: 71.9 ± 8.84 years, gender: five males and three females, time postmortem: 27.6 ± 16.3 h, cultivation time before transfection: 67.3 ± 24.0 days) were transfected with an increasing ratio (1:1–1:24) of SB100X transposase to BDNF transposon plasmid at a total concentration of 0.5 µg, plus one control without the addition of plasmid DNA (Co). Cultures were terminated 36.8 ± 10.5 days after transfection. Cells were used for isolation of total RNA. Medium used for ELISA was added to the cells in a defined volume 24 h before culture termination. ( a ) BDNF gene expression was analyzed by qPCR. Total BDNF expression in transfected cultures was related to the expression in Co cultures, which was set to 1 (dashed line). An increase in BDNF expression was observed in the transfected cultures (ns: not significant, * p < 0.05, one sample t test). ( b ) BDNF secretion was quantified by ELISA using precisely defined cell culture supernatants. Values of the transfected cultures were higher than those of the Co cultures, although not significantly (Brown-Forsythe and Welch ANOVA tests with Dunnett’s T3 multiple comparisons test). Data are presented as box and whisker plots (whiskers: minimum to maximum, mean values indicated by +). Outliers were evaluated using the ROUT method (Q = 1%, indicated by red open circles).

Techniques Used: Transfection, Plasmid Preparation, Concentration Assay, Isolation, Enzyme-linked Immunosorbent Assay, Expressing, Cell Culture, Whisker Assay

Transposition efficiency in primary hRPE cells transfected with a defined ratio of SB100X transposase to BDNF transposon plasmid. In each experiment ( n = 4), two to eight transfections were performed using 5 × 10 4 or 1 × 10 5 cells from individual donors (donor age: 66.5 ± 9.95 years, gender: four males, time postmortem: 29.8 ± 28.0 h, cultivation time before transfection: 74.8 ± 51.1 days) and a mixture of 0.03 µg SB100X transposase and 0.47 µg BDNF transposon plasmid (BDNF + ), plus one or two controls without the addition of plasmid DNA (Co). Cultures were terminated 22 days after transfection. Cells were used for isolation of total RNA. Medium used for ELISA was added to the cells in a defined volume 24 h before culture termination. ( a ) BDNF gene expression was analyzed by qPCR. Endogenous and total BDNF expression in BDNF + cultures were related to the respective expression in Co cultures, which were set to 1 (dashed line). An increase in total BDNF expression was observed in the BDNF + cultures (ns: not significant, summarized data: **** p < 0.0001, unpaired t test with Welch’s correction; single experiments: * p = 0.0418, ** p = 0.0023, 2way ANOVA with Šídák’s multiple comparisons test). ( b ) BDNF secretion was quantified by ELISA using precisely defined cell culture supernatants. Values of the BDNF + cultures were higher than those of the Co cultures (ns: not significant, summarized data: * p = 0.0333, unpaired t test with Welch’s correction; single experiments: ** p = 0.0014, 2way ANOVA with Šídák’s multiple comparisons test). Data are presented as box and whisker plots (whiskers: minimum to maximum, mean values indicated by +) and as individual values per experiment (line at mean). Outliers were evaluated using the ROUT method (Q = 1%, as indicated by red open circles).
Figure Legend Snippet: Transposition efficiency in primary hRPE cells transfected with a defined ratio of SB100X transposase to BDNF transposon plasmid. In each experiment ( n = 4), two to eight transfections were performed using 5 × 10 4 or 1 × 10 5 cells from individual donors (donor age: 66.5 ± 9.95 years, gender: four males, time postmortem: 29.8 ± 28.0 h, cultivation time before transfection: 74.8 ± 51.1 days) and a mixture of 0.03 µg SB100X transposase and 0.47 µg BDNF transposon plasmid (BDNF + ), plus one or two controls without the addition of plasmid DNA (Co). Cultures were terminated 22 days after transfection. Cells were used for isolation of total RNA. Medium used for ELISA was added to the cells in a defined volume 24 h before culture termination. ( a ) BDNF gene expression was analyzed by qPCR. Endogenous and total BDNF expression in BDNF + cultures were related to the respective expression in Co cultures, which were set to 1 (dashed line). An increase in total BDNF expression was observed in the BDNF + cultures (ns: not significant, summarized data: **** p < 0.0001, unpaired t test with Welch’s correction; single experiments: * p = 0.0418, ** p = 0.0023, 2way ANOVA with Šídák’s multiple comparisons test). ( b ) BDNF secretion was quantified by ELISA using precisely defined cell culture supernatants. Values of the BDNF + cultures were higher than those of the Co cultures (ns: not significant, summarized data: * p = 0.0333, unpaired t test with Welch’s correction; single experiments: ** p = 0.0014, 2way ANOVA with Šídák’s multiple comparisons test). Data are presented as box and whisker plots (whiskers: minimum to maximum, mean values indicated by +) and as individual values per experiment (line at mean). Outliers were evaluated using the ROUT method (Q = 1%, as indicated by red open circles).

Techniques Used: Transfection, Plasmid Preparation, Isolation, Enzyme-linked Immunosorbent Assay, Expressing, Cell Culture, Whisker Assay


Structured Review

R&D Systems human free bdnf quantikine elisa kit
Efficiency of transgene expression in the CreVP64/lox71 system. ( A ) Flow cytometry of cells transfected with constructs encoding PKA-spark. *, The percentage of bright HEK293T cells among all GFP-positive cells. ( B ) Fluorescence microscopy of cells transfected with constructs encoding PKA-spark. ( C , F ) Fold enhancement (to “before induction” level) of PKA-spark ( C ) and <t>BDNF</t> ( F ) mRNAs in HEK293T cells 48 h after induction. ( D ) PKA-spark sensor fluorescence intensity of cells transfected with pVax1-lox71 3/6 -C-PKA-spark with or without induction. **, p < 0.05, n ≥ 30,000. Data are presented as the median (25%; 75%). ( E ) The secretion of human BDNF by transfected cells measured by <t>ELISA</t> 48 h after induction.
Human Free Bdnf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Price from $9.99 to $1999.99
human free bdnf quantikine elisa kit - by Bioz Stars, 2023-01
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1) Product Images from "A Novel Cre/lox71-Based System for Inducible Expression of Recombinant Proteins and Genome Editing"

Article Title: A Novel Cre/lox71-Based System for Inducible Expression of Recombinant Proteins and Genome Editing

Journal: Cells

doi: 10.3390/cells11142141

Efficiency of transgene expression in the CreVP64/lox71 system. ( A ) Flow cytometry of cells transfected with constructs encoding PKA-spark. *, The percentage of bright HEK293T cells among all GFP-positive cells. ( B ) Fluorescence microscopy of cells transfected with constructs encoding PKA-spark. ( C , F ) Fold enhancement (to “before induction” level) of PKA-spark ( C ) and BDNF ( F ) mRNAs in HEK293T cells 48 h after induction. ( D ) PKA-spark sensor fluorescence intensity of cells transfected with pVax1-lox71 3/6 -C-PKA-spark with or without induction. **, p < 0.05, n ≥ 30,000. Data are presented as the median (25%; 75%). ( E ) The secretion of human BDNF by transfected cells measured by ELISA 48 h after induction.
Figure Legend Snippet: Efficiency of transgene expression in the CreVP64/lox71 system. ( A ) Flow cytometry of cells transfected with constructs encoding PKA-spark. *, The percentage of bright HEK293T cells among all GFP-positive cells. ( B ) Fluorescence microscopy of cells transfected with constructs encoding PKA-spark. ( C , F ) Fold enhancement (to “before induction” level) of PKA-spark ( C ) and BDNF ( F ) mRNAs in HEK293T cells 48 h after induction. ( D ) PKA-spark sensor fluorescence intensity of cells transfected with pVax1-lox71 3/6 -C-PKA-spark with or without induction. **, p < 0.05, n ≥ 30,000. Data are presented as the median (25%; 75%). ( E ) The secretion of human BDNF by transfected cells measured by ELISA 48 h after induction.

Techniques Used: Expressing, Flow Cytometry, Transfection, Construct, Fluorescence, Microscopy, Enzyme-linked Immunosorbent Assay


Structured Review

R&D Systems human free bdnf quantikine elisa kit
Human Free Bdnf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human free bdnf quantikine elisa kit/product/R&D Systems
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
human free bdnf quantikine elisa kit - by Bioz Stars, 2023-01
86/100 stars

Images


Structured Review

R&D Systems human free bdnf quantikine elisa kit
Human Free Bdnf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human free bdnf quantikine elisa kit/product/R&D Systems
Average 86 stars, based on 1 article reviews
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R&D Systems human free bdnf quantikine elisa kit
Human Free Bdnf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human free bdnf quantikine elisa kit/product/R&D Systems
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
human free bdnf quantikine elisa kit - by Bioz Stars, 2023-01
86/100 stars

Images


Structured Review

R&D Systems o c human free bdnf quantikine elisa kit
O C Human Free Bdnf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/o c human free bdnf quantikine elisa kit/product/R&D Systems
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
o c human free bdnf quantikine elisa kit - by Bioz Stars, 2023-01
86/100 stars

Images


Structured Review

R&D Systems 80o c human free bdnf quantikine elisa kit
80o C Human Free Bdnf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/80o c human free bdnf quantikine elisa kit/product/R&D Systems
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
80o c human free bdnf quantikine elisa kit - by Bioz Stars, 2023-01
86/100 stars

Images


Structured Review

R&D Systems human free bdnf quantikine elisa kit
Characterization of <t>BDNF-</t> overexpressing engineered mesenchymal stem cells (eMSCs). ( A ) Morphology of BDNF-eMSCs. ( B ) Flow cytometry analyses show that BDNF-eMSCs express specific markers for MSCs such as CD44, CD105, and CD73. N = three per group ( C ) Cell proliferation rate of BDNF-eMSCs. This result is representative of three independent experiments. ( D ) BDNF secretion of distinct passages of BDNF-eMSCs measured by <t>ELISA.</t>
Human Free Bdnf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human free bdnf quantikine elisa kit/product/R&D Systems
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
human free bdnf quantikine elisa kit - by Bioz Stars, 2023-01
86/100 stars

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1) Product Images from "BDNF-Overexpressing Engineered Mesenchymal Stem Cells Enhances Their Therapeutic Efficacy against Severe Neonatal Hypoxic Ischemic Brain Injury"

Article Title: BDNF-Overexpressing Engineered Mesenchymal Stem Cells Enhances Their Therapeutic Efficacy against Severe Neonatal Hypoxic Ischemic Brain Injury

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms222111395

Characterization of BDNF- overexpressing engineered mesenchymal stem cells (eMSCs). ( A ) Morphology of BDNF-eMSCs. ( B ) Flow cytometry analyses show that BDNF-eMSCs express specific markers for MSCs such as CD44, CD105, and CD73. N = three per group ( C ) Cell proliferation rate of BDNF-eMSCs. This result is representative of three independent experiments. ( D ) BDNF secretion of distinct passages of BDNF-eMSCs measured by ELISA.
Figure Legend Snippet: Characterization of BDNF- overexpressing engineered mesenchymal stem cells (eMSCs). ( A ) Morphology of BDNF-eMSCs. ( B ) Flow cytometry analyses show that BDNF-eMSCs express specific markers for MSCs such as CD44, CD105, and CD73. N = three per group ( C ) Cell proliferation rate of BDNF-eMSCs. This result is representative of three independent experiments. ( D ) BDNF secretion of distinct passages of BDNF-eMSCs measured by ELISA.

Techniques Used: Flow Cytometry, Enzyme-linked Immunosorbent Assay

Neuroprotective efficacy of BDNF-eMSCs in vitro oxygen–glucose deprivation (OGD)-induced primary cultured rat cortical neurons. Cell viability, which is expressed as relative proliferation rate (%) to normal control group, was assessed with the CCK-8 assay in OGD-induced cultured neurons after co-treatment with BDNF-eMSCs, at doses of approximately 1 × 10 3 , 1 × 10 4 , 5 × 10 4 , 1 × 10 5 , 1 × 10 6 cells per 1 mL, ( A ) and with naïve MSCs at doses of approximately 1 × 10 3 , 1 × 10 4 , 5 × 10 4 , 1 × 10 5 , 1 × 10 6 cells per 1 mL ( B ). ( C ) Western blotting and ELISA assay for BDNF expression from the culture media of naïve MSCs and BDNF-eMSCs. ( D ) Cytotoxicity was evaluated with cell viability, expressed as a relative proliferation rate (%) to the normal control group, relative lactate dehydrogenase (LDH) release (%) to the positive control (100% fully killed cells), malondialdehyde (MDA) level in neuronal cells after OGD induction with/without co-treatment of naïve MSCs or BDNF-eMSCs at a dose of 1 × 10 5 cells per 1 mL. ( E ) The number of terminal deoxynucleotidyl transferase UTP nick end labeling (TUNEL)-positive cells evaluated in OGD-induced rat cortical neurons after co-culture with naïve MSCs or BDNF-eMSCs at dose of 1 × 10 5 cells per 1 mL. Data are expressed as the mean ± SD.
Figure Legend Snippet: Neuroprotective efficacy of BDNF-eMSCs in vitro oxygen–glucose deprivation (OGD)-induced primary cultured rat cortical neurons. Cell viability, which is expressed as relative proliferation rate (%) to normal control group, was assessed with the CCK-8 assay in OGD-induced cultured neurons after co-treatment with BDNF-eMSCs, at doses of approximately 1 × 10 3 , 1 × 10 4 , 5 × 10 4 , 1 × 10 5 , 1 × 10 6 cells per 1 mL, ( A ) and with naïve MSCs at doses of approximately 1 × 10 3 , 1 × 10 4 , 5 × 10 4 , 1 × 10 5 , 1 × 10 6 cells per 1 mL ( B ). ( C ) Western blotting and ELISA assay for BDNF expression from the culture media of naïve MSCs and BDNF-eMSCs. ( D ) Cytotoxicity was evaluated with cell viability, expressed as a relative proliferation rate (%) to the normal control group, relative lactate dehydrogenase (LDH) release (%) to the positive control (100% fully killed cells), malondialdehyde (MDA) level in neuronal cells after OGD induction with/without co-treatment of naïve MSCs or BDNF-eMSCs at a dose of 1 × 10 5 cells per 1 mL. ( E ) The number of terminal deoxynucleotidyl transferase UTP nick end labeling (TUNEL)-positive cells evaluated in OGD-induced rat cortical neurons after co-culture with naïve MSCs or BDNF-eMSCs at dose of 1 × 10 5 cells per 1 mL. Data are expressed as the mean ± SD.

Techniques Used: In Vitro, Cell Culture, CCK-8 Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Positive Control, End Labeling, TUNEL Assay, Co-Culture Assay

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    R&D Systems human free bdnf quantikine elisa kit
    Retinal GFP and <t>BDNF</t> expression after intravitreal injection of tm-scAAV2-GFP and tm-scAAV2-BDNF. A : Fluorescence microscopic images of cross sections treated with tm-scAAV2-GFP and NMDA. Green: GFP, Blue: DAPI. Scale bar represents 50 μm. B : Relative RNA expression levels of BDNF analyzed by the comparative threshold cycle method. GAPDH was used to normalize gene expressions (n = 6 in each group, *p<0.05 for a two-tailed Mann–Whitney U test). C : Protein expression levels of BDNF after intravitreal injection of tm-scAAV2-BDNF, as analyzed by <t>ELISA</t> (n = 4 in each group, *p<0.05 for a two-tailed Mann–Whitney U test).
    Human Free Bdnf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human free bdnf quantikine elisa kit/product/R&D Systems
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human free bdnf quantikine elisa kit - by Bioz Stars, 2023-01
    86/100 stars
      Buy from Supplier

    86
    R&D Systems quantikine human free bdnf elisa kit
    Retinal GFP and <t>BDNF</t> expression after intravitreal injection of tm-scAAV2-GFP and tm-scAAV2-BDNF. A : Fluorescence microscopic images of cross sections treated with tm-scAAV2-GFP and NMDA. Green: GFP, Blue: DAPI. Scale bar represents 50 μm. B : Relative RNA expression levels of BDNF analyzed by the comparative threshold cycle method. GAPDH was used to normalize gene expressions (n = 6 in each group, *p<0.05 for a two-tailed Mann–Whitney U test). C : Protein expression levels of BDNF after intravitreal injection of tm-scAAV2-BDNF, as analyzed by <t>ELISA</t> (n = 4 in each group, *p<0.05 for a two-tailed Mann–Whitney U test).
    Quantikine Human Free Bdnf Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantikine human free bdnf elisa kit/product/R&D Systems
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    quantikine human free bdnf elisa kit - by Bioz Stars, 2023-01
    86/100 stars
      Buy from Supplier

    86
    R&D Systems o c human free bdnf quantikine elisa kit
    Retinal GFP and <t>BDNF</t> expression after intravitreal injection of tm-scAAV2-GFP and tm-scAAV2-BDNF. A : Fluorescence microscopic images of cross sections treated with tm-scAAV2-GFP and NMDA. Green: GFP, Blue: DAPI. Scale bar represents 50 μm. B : Relative RNA expression levels of BDNF analyzed by the comparative threshold cycle method. GAPDH was used to normalize gene expressions (n = 6 in each group, *p<0.05 for a two-tailed Mann–Whitney U test). C : Protein expression levels of BDNF after intravitreal injection of tm-scAAV2-BDNF, as analyzed by <t>ELISA</t> (n = 4 in each group, *p<0.05 for a two-tailed Mann–Whitney U test).
    O C Human Free Bdnf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/o c human free bdnf quantikine elisa kit/product/R&D Systems
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    o c human free bdnf quantikine elisa kit - by Bioz Stars, 2023-01
    86/100 stars
      Buy from Supplier

    86
    R&D Systems 80o c human free bdnf quantikine elisa kit
    Retinal GFP and <t>BDNF</t> expression after intravitreal injection of tm-scAAV2-GFP and tm-scAAV2-BDNF. A : Fluorescence microscopic images of cross sections treated with tm-scAAV2-GFP and NMDA. Green: GFP, Blue: DAPI. Scale bar represents 50 μm. B : Relative RNA expression levels of BDNF analyzed by the comparative threshold cycle method. GAPDH was used to normalize gene expressions (n = 6 in each group, *p<0.05 for a two-tailed Mann–Whitney U test). C : Protein expression levels of BDNF after intravitreal injection of tm-scAAV2-BDNF, as analyzed by <t>ELISA</t> (n = 4 in each group, *p<0.05 for a two-tailed Mann–Whitney U test).
    80o C Human Free Bdnf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/80o c human free bdnf quantikine elisa kit/product/R&D Systems
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    80o c human free bdnf quantikine elisa kit - by Bioz Stars, 2023-01
    86/100 stars
      Buy from Supplier

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    Retinal GFP and BDNF expression after intravitreal injection of tm-scAAV2-GFP and tm-scAAV2-BDNF. A : Fluorescence microscopic images of cross sections treated with tm-scAAV2-GFP and NMDA. Green: GFP, Blue: DAPI. Scale bar represents 50 μm. B : Relative RNA expression levels of BDNF analyzed by the comparative threshold cycle method. GAPDH was used to normalize gene expressions (n = 6 in each group, *p<0.05 for a two-tailed Mann–Whitney U test). C : Protein expression levels of BDNF after intravitreal injection of tm-scAAV2-BDNF, as analyzed by ELISA (n = 4 in each group, *p<0.05 for a two-tailed Mann–Whitney U test).

    Journal: Molecular Vision

    Article Title: Tyrosine triple mutated AAV2-BDNF gene therapy in an inner retinal injury model induced by intravitreal injection of N –methyl-D-aspartate (NMDA)

    doi:

    Figure Lengend Snippet: Retinal GFP and BDNF expression after intravitreal injection of tm-scAAV2-GFP and tm-scAAV2-BDNF. A : Fluorescence microscopic images of cross sections treated with tm-scAAV2-GFP and NMDA. Green: GFP, Blue: DAPI. Scale bar represents 50 μm. B : Relative RNA expression levels of BDNF analyzed by the comparative threshold cycle method. GAPDH was used to normalize gene expressions (n = 6 in each group, *p<0.05 for a two-tailed Mann–Whitney U test). C : Protein expression levels of BDNF after intravitreal injection of tm-scAAV2-BDNF, as analyzed by ELISA (n = 4 in each group, *p<0.05 for a two-tailed Mann–Whitney U test).

    Article Snippet: Homogenized retinas were examined using the Human Free BDNF Quantikine ELISA Kit (R&D Systems, Inc., MN) in accordance with the product protocol.

    Techniques: Expressing, Injection, Fluorescence, RNA Expression, Two Tailed Test, MANN-WHITNEY, Enzyme-linked Immunosorbent Assay