ly294002  (Echelon Biosciences)


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    Echelon Biosciences ly294002
    Standard plot of absorbance at 450 nm versus (A) log pmol PI(4,5)P2 and (C) log pmol PI3P. B Percentage change in PI(4,5)P2 content after BL irradiation in control (10 mM PIPES), neomycin (250 µM) and U73122 (25 µM) samples. D Percentage change in PI3P level after BL irradiation in control (10 mM PIPES), WM (10 µM) and <t>LY294002</t> (200 µM). The vertical bars represent SE of 3 independent experiments.
    Ly294002, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Phosphoinositides Play Differential Roles in Regulating Phototropin1- and Phototropin2-Mediated Chloroplast Movements in Arabidopsis"

    Article Title: Phosphoinositides Play Differential Roles in Regulating Phototropin1- and Phototropin2-Mediated Chloroplast Movements in Arabidopsis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0055393

    Standard plot of absorbance at 450 nm versus (A) log pmol PI(4,5)P2 and (C) log pmol PI3P. B Percentage change in PI(4,5)P2 content after BL irradiation in control (10 mM PIPES), neomycin (250 µM) and U73122 (25 µM) samples. D Percentage change in PI3P level after BL irradiation in control (10 mM PIPES), WM (10 µM) and LY294002 (200 µM). The vertical bars represent SE of 3 independent experiments.
    Figure Legend Snippet: Standard plot of absorbance at 450 nm versus (A) log pmol PI(4,5)P2 and (C) log pmol PI3P. B Percentage change in PI(4,5)P2 content after BL irradiation in control (10 mM PIPES), neomycin (250 µM) and U73122 (25 µM) samples. D Percentage change in PI3P level after BL irradiation in control (10 mM PIPES), WM (10 µM) and LY294002 (200 µM). The vertical bars represent SE of 3 independent experiments.

    Techniques Used: Irradiation

    Leaves were infiltrated with ( A and B ) WM (10 µM) or ( C and D ) LY294002 (200 µM) and incubated for 1.5 h under darkness. Parameters of chloroplast responses: (A, C) amplitudes and (B, D) kinetics after treatment with inhibitors. Final readings are presented as mean ± SE (n = 10) from 2 independent experiments.
    Figure Legend Snippet: Leaves were infiltrated with ( A and B ) WM (10 µM) or ( C and D ) LY294002 (200 µM) and incubated for 1.5 h under darkness. Parameters of chloroplast responses: (A, C) amplitudes and (B, D) kinetics after treatment with inhibitors. Final readings are presented as mean ± SE (n = 10) from 2 independent experiments.

    Techniques Used: Incubation

    A Leaf discs were illuminated with 100 µmol m −2 s −1 of BL for 5 s. B and C Leaf discs treated with (B) 10/25 µM U73122 or U73343 and (C) 10 µM WM, 50 µM WM or 200 µM LY294002 60 min before the measurements and then irradiated with 100 µmol m −2 s −1 BL for 5 s. The vertical bars represent SE of 20 independent experiments for each. The arrow shows the onset of BL pulse.
    Figure Legend Snippet: A Leaf discs were illuminated with 100 µmol m −2 s −1 of BL for 5 s. B and C Leaf discs treated with (B) 10/25 µM U73122 or U73343 and (C) 10 µM WM, 50 µM WM or 200 µM LY294002 60 min before the measurements and then irradiated with 100 µmol m −2 s −1 BL for 5 s. The vertical bars represent SE of 20 independent experiments for each. The arrow shows the onset of BL pulse.

    Techniques Used: Irradiation

    ly294002  (Echelon Biosciences)


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    Echelon Biosciences ly294002
    Standard plot of absorbance at 450 nm versus (A) log pmol PI(4,5)P2 and (C) log pmol PI3P. B Percentage change in PI(4,5)P2 content after BL irradiation in control (10 mM PIPES), neomycin (250 µM) and U73122 (25 µM) samples. D Percentage change in PI3P level after BL irradiation in control (10 mM PIPES), WM (10 µM) and <t>LY294002</t> (200 µM). The vertical bars represent SE of 3 independent experiments.
    Ly294002, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Images

    1) Product Images from "Phosphoinositides Play Differential Roles in Regulating Phototropin1- and Phototropin2-Mediated Chloroplast Movements in Arabidopsis"

    Article Title: Phosphoinositides Play Differential Roles in Regulating Phototropin1- and Phototropin2-Mediated Chloroplast Movements in Arabidopsis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0055393

    Standard plot of absorbance at 450 nm versus (A) log pmol PI(4,5)P2 and (C) log pmol PI3P. B Percentage change in PI(4,5)P2 content after BL irradiation in control (10 mM PIPES), neomycin (250 µM) and U73122 (25 µM) samples. D Percentage change in PI3P level after BL irradiation in control (10 mM PIPES), WM (10 µM) and LY294002 (200 µM). The vertical bars represent SE of 3 independent experiments.
    Figure Legend Snippet: Standard plot of absorbance at 450 nm versus (A) log pmol PI(4,5)P2 and (C) log pmol PI3P. B Percentage change in PI(4,5)P2 content after BL irradiation in control (10 mM PIPES), neomycin (250 µM) and U73122 (25 µM) samples. D Percentage change in PI3P level after BL irradiation in control (10 mM PIPES), WM (10 µM) and LY294002 (200 µM). The vertical bars represent SE of 3 independent experiments.

    Techniques Used: Irradiation

    Leaves were infiltrated with ( A and B ) WM (10 µM) or ( C and D ) LY294002 (200 µM) and incubated for 1.5 h under darkness. Parameters of chloroplast responses: (A, C) amplitudes and (B, D) kinetics after treatment with inhibitors. Final readings are presented as mean ± SE (n = 10) from 2 independent experiments.
    Figure Legend Snippet: Leaves were infiltrated with ( A and B ) WM (10 µM) or ( C and D ) LY294002 (200 µM) and incubated for 1.5 h under darkness. Parameters of chloroplast responses: (A, C) amplitudes and (B, D) kinetics after treatment with inhibitors. Final readings are presented as mean ± SE (n = 10) from 2 independent experiments.

    Techniques Used: Incubation

    A Leaf discs were illuminated with 100 µmol m −2 s −1 of BL for 5 s. B and C Leaf discs treated with (B) 10/25 µM U73122 or U73343 and (C) 10 µM WM, 50 µM WM or 200 µM LY294002 60 min before the measurements and then irradiated with 100 µmol m −2 s −1 BL for 5 s. The vertical bars represent SE of 20 independent experiments for each. The arrow shows the onset of BL pulse.
    Figure Legend Snippet: A Leaf discs were illuminated with 100 µmol m −2 s −1 of BL for 5 s. B and C Leaf discs treated with (B) 10/25 µM U73122 or U73343 and (C) 10 µM WM, 50 µM WM or 200 µM LY294002 60 min before the measurements and then irradiated with 100 µmol m −2 s −1 BL for 5 s. The vertical bars represent SE of 20 independent experiments for each. The arrow shows the onset of BL pulse.

    Techniques Used: Irradiation

    ly294002  (Echelon Biosciences)


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    Echelon Biosciences ly294002
    A. PI3K inhibitors reduce plaque size in BSC40 cells. Cells were infected with 100 PFU of vaccinia virus (VV), strain WR. Inhibitors were added one hour post viral entry. First row: <t>LY294002,</t> 20 µM; B0304, 20 µM; AS1, 50 µM; AS2, 50 µM (LY294002 and B0304 are in 0.2% DMSO and AS1 and AS2 are in 0.5% DMSO); 0.5% DMSO alone. Insets show representative plaques from 50 µM AS1 and 0.5% DMSO treated wells. Second row: 10 µM Wortmannin in 0.1% DMSO did not appear to reduce VV plaque size; 0.1% DMSO control. Results are from three independent trials. B. p85WT and p85α −/− cells form two times more plaques than p85α −/− β −/− cells. Cells were infected with equal amounts of virus and monolayers were stained with crystal violet 48 hours post infection. Results are from eight independent trials. C. PI3K inhibitors do not decrease early viral protein expression. BSC40 cells were infected at pH 7.4 and MOI = 5 with VV and PI3K inhibitors were added 1 hour post-viral entry and allowed to infect for an additional 2 hours. Proteins were subjected to western analysis by anti-E3 mAb and anti-Tubulin mAb. For quantification (right) bands were normalized to tubulin loading control and data is expressed as the fold change relative to DMSO-treated samples. D. Early protein expression is reduced in p85-deficient cells. Cells were infected at an MOI = 5 with VV at pH = 7.4 and analyzed as in C. E. Early protein expression is reduced in p85-deficient cells. Cells were infected at an MOI = 5 with VV. Virus was bound to cells at 4°C, after which cells were moved to 37°C with PBS, pH = 5. Following a 2 hour infection, equal amounts of protein were subjected to western analysis with anti-E3 mAb or anti-Tubulin, and quantitated as in C.
    Ly294002, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ly294002 - by Bioz Stars, 2024-03
    93/100 stars

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    1) Product Images from "Multiple Phosphatidylinositol 3-Kinases Regulate Vaccinia Virus Morphogenesis"

    Article Title: Multiple Phosphatidylinositol 3-Kinases Regulate Vaccinia Virus Morphogenesis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0010884

    A. PI3K inhibitors reduce plaque size in BSC40 cells. Cells were infected with 100 PFU of vaccinia virus (VV), strain WR. Inhibitors were added one hour post viral entry. First row: LY294002, 20 µM; B0304, 20 µM; AS1, 50 µM; AS2, 50 µM (LY294002 and B0304 are in 0.2% DMSO and AS1 and AS2 are in 0.5% DMSO); 0.5% DMSO alone. Insets show representative plaques from 50 µM AS1 and 0.5% DMSO treated wells. Second row: 10 µM Wortmannin in 0.1% DMSO did not appear to reduce VV plaque size; 0.1% DMSO control. Results are from three independent trials. B. p85WT and p85α −/− cells form two times more plaques than p85α −/− β −/− cells. Cells were infected with equal amounts of virus and monolayers were stained with crystal violet 48 hours post infection. Results are from eight independent trials. C. PI3K inhibitors do not decrease early viral protein expression. BSC40 cells were infected at pH 7.4 and MOI = 5 with VV and PI3K inhibitors were added 1 hour post-viral entry and allowed to infect for an additional 2 hours. Proteins were subjected to western analysis by anti-E3 mAb and anti-Tubulin mAb. For quantification (right) bands were normalized to tubulin loading control and data is expressed as the fold change relative to DMSO-treated samples. D. Early protein expression is reduced in p85-deficient cells. Cells were infected at an MOI = 5 with VV at pH = 7.4 and analyzed as in C. E. Early protein expression is reduced in p85-deficient cells. Cells were infected at an MOI = 5 with VV. Virus was bound to cells at 4°C, after which cells were moved to 37°C with PBS, pH = 5. Following a 2 hour infection, equal amounts of protein were subjected to western analysis with anti-E3 mAb or anti-Tubulin, and quantitated as in C.
    Figure Legend Snippet: A. PI3K inhibitors reduce plaque size in BSC40 cells. Cells were infected with 100 PFU of vaccinia virus (VV), strain WR. Inhibitors were added one hour post viral entry. First row: LY294002, 20 µM; B0304, 20 µM; AS1, 50 µM; AS2, 50 µM (LY294002 and B0304 are in 0.2% DMSO and AS1 and AS2 are in 0.5% DMSO); 0.5% DMSO alone. Insets show representative plaques from 50 µM AS1 and 0.5% DMSO treated wells. Second row: 10 µM Wortmannin in 0.1% DMSO did not appear to reduce VV plaque size; 0.1% DMSO control. Results are from three independent trials. B. p85WT and p85α −/− cells form two times more plaques than p85α −/− β −/− cells. Cells were infected with equal amounts of virus and monolayers were stained with crystal violet 48 hours post infection. Results are from eight independent trials. C. PI3K inhibitors do not decrease early viral protein expression. BSC40 cells were infected at pH 7.4 and MOI = 5 with VV and PI3K inhibitors were added 1 hour post-viral entry and allowed to infect for an additional 2 hours. Proteins were subjected to western analysis by anti-E3 mAb and anti-Tubulin mAb. For quantification (right) bands were normalized to tubulin loading control and data is expressed as the fold change relative to DMSO-treated samples. D. Early protein expression is reduced in p85-deficient cells. Cells were infected at an MOI = 5 with VV at pH = 7.4 and analyzed as in C. E. Early protein expression is reduced in p85-deficient cells. Cells were infected at an MOI = 5 with VV. Virus was bound to cells at 4°C, after which cells were moved to 37°C with PBS, pH = 5. Following a 2 hour infection, equal amounts of protein were subjected to western analysis with anti-E3 mAb or anti-Tubulin, and quantitated as in C.

    Techniques Used: Infection, Staining, Expressing, Western Blot

    50% Inhibitory Concentration (IC 50 ) of PI3K inhibitors added post-adsorption to poxvirus-infected monolayers.
    Figure Legend Snippet: 50% Inhibitory Concentration (IC 50 ) of PI3K inhibitors added post-adsorption to poxvirus-infected monolayers.

    Techniques Used: Concentration Assay

    ly294002  (Echelon Biosciences)


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    Echelon Biosciences ly294002
    Schematic diagram of the method for exploring the synergistic effects of ILK and <t>PI3K</t> in the role of static compressive stress on autophagy of human periodontal ligament cells (hPDLCs). hPDLCs were obtained from the human periodontal ligament and divided into untreated, ILK silencing, and PI3K inhibitory groups. The compressive force of hPDLCs was simulated by applying gravity on the collagen-alginate hydrogel in vitro . The effects of mechanical stress on hPDLC autophagy and the potential modulatory mechanisms were then demonstrated by a series of corresponding assays. ILK, integrin-linked kinase; PI3K, phosphatidylinositol 3 kinase.
    Ly294002, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Role of integrin-linked kinase in static compressive stress-induced autophagy via phosphatidylinositol 3 kinase in human periodontal ligament cells"

    Article Title: Role of integrin-linked kinase in static compressive stress-induced autophagy via phosphatidylinositol 3 kinase in human periodontal ligament cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2021.5000

    Schematic diagram of the method for exploring the synergistic effects of ILK and PI3K in the role of static compressive stress on autophagy of human periodontal ligament cells (hPDLCs). hPDLCs were obtained from the human periodontal ligament and divided into untreated, ILK silencing, and PI3K inhibitory groups. The compressive force of hPDLCs was simulated by applying gravity on the collagen-alginate hydrogel in vitro . The effects of mechanical stress on hPDLC autophagy and the potential modulatory mechanisms were then demonstrated by a series of corresponding assays. ILK, integrin-linked kinase; PI3K, phosphatidylinositol 3 kinase.
    Figure Legend Snippet: Schematic diagram of the method for exploring the synergistic effects of ILK and PI3K in the role of static compressive stress on autophagy of human periodontal ligament cells (hPDLCs). hPDLCs were obtained from the human periodontal ligament and divided into untreated, ILK silencing, and PI3K inhibitory groups. The compressive force of hPDLCs was simulated by applying gravity on the collagen-alginate hydrogel in vitro . The effects of mechanical stress on hPDLC autophagy and the potential modulatory mechanisms were then demonstrated by a series of corresponding assays. ILK, integrin-linked kinase; PI3K, phosphatidylinositol 3 kinase.

    Techniques Used: In Vitro

    Loading static stress after PI3K inhibition. (A) Optical density value under different concentrations of drugs. (B) Protein expression level of AKT, P-AKT (Ser473) and GAPDH. (C) The ratio of P-AKT/total AKT level. (D) ILK gene expression. No significant change was found between groups (P>0.05). (E) Beclin-1 ( BECN1 ) gene expression was slightly increased at 15 min (P<0.05), while no significant change was observed in other groups (P>0.05). (F) ATG-5 gene expression remained unchanged between groups (P>0.05). (G) The expression of LC3 (red fluorescence: LC3; blue fluorescence: DAPI; scale bar, 200 µ m). (H) The ratio of LC3II/LC3I. Data are presented as mean ± standard error of the mean. * P<0.05 and ** P<0.01, significant differences vs. the 0 min group; ns, not significant (P>0.05). PI3K, phosphatidylinositol 3 kinase; ILK, integrin-linked kinase; DAPI, 4-6-diamidino-2-phenylindole.
    Figure Legend Snippet: Loading static stress after PI3K inhibition. (A) Optical density value under different concentrations of drugs. (B) Protein expression level of AKT, P-AKT (Ser473) and GAPDH. (C) The ratio of P-AKT/total AKT level. (D) ILK gene expression. No significant change was found between groups (P>0.05). (E) Beclin-1 ( BECN1 ) gene expression was slightly increased at 15 min (P<0.05), while no significant change was observed in other groups (P>0.05). (F) ATG-5 gene expression remained unchanged between groups (P>0.05). (G) The expression of LC3 (red fluorescence: LC3; blue fluorescence: DAPI; scale bar, 200 µ m). (H) The ratio of LC3II/LC3I. Data are presented as mean ± standard error of the mean. * P<0.05 and ** P<0.01, significant differences vs. the 0 min group; ns, not significant (P>0.05). PI3K, phosphatidylinositol 3 kinase; ILK, integrin-linked kinase; DAPI, 4-6-diamidino-2-phenylindole.

    Techniques Used: Inhibition, Expressing, Fluorescence

    inhibitors  (Echelon Biosciences)


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    Echelon Biosciences inhibitors
    Inhibitors, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ly294002  (Echelon Biosciences)


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    Echelon Biosciences ly294002
    A–C A mass ELISA was used to detect specific PIP species after exposure to anti‐FcγRII/III (2.4G2) with or without 3‐a‐aminocholestane (SHIP 1 inhibitor), SF1670 (PTEN inhibitor) and <t>LY294002</t> (PI‐3K inhibitor). In M‐MOP cells (left panel) and primary microglia (right panel), PI(4,5)P 2 (A), PI(3,4)P 2 (B) and PIP 3 (C) were detected. All data show the mean ± SD of three independent experiments and were analysed by two‐way ANOVA (log‐transformed data were used in C) with Sidak’s multiple comparison tests performed. * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001 (blue: P522 and red: R522).
    Ly294002, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "PIP2 depletion and altered endocytosis caused by expression of Alzheimer's disease‐protective variant PLCγ2 R522"

    Article Title: PIP2 depletion and altered endocytosis caused by expression of Alzheimer's disease‐protective variant PLCγ2 R522

    Journal: The EMBO Journal

    doi: 10.15252/embj.2020105603

    A–C A mass ELISA was used to detect specific PIP species after exposure to anti‐FcγRII/III (2.4G2) with or without 3‐a‐aminocholestane (SHIP 1 inhibitor), SF1670 (PTEN inhibitor) and LY294002 (PI‐3K inhibitor). In M‐MOP cells (left panel) and primary microglia (right panel), PI(4,5)P 2 (A), PI(3,4)P 2 (B) and PIP 3 (C) were detected. All data show the mean ± SD of three independent experiments and were analysed by two‐way ANOVA (log‐transformed data were used in C) with Sidak’s multiple comparison tests performed. * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001 (blue: P522 and red: R522).
    Figure Legend Snippet: A–C A mass ELISA was used to detect specific PIP species after exposure to anti‐FcγRII/III (2.4G2) with or without 3‐a‐aminocholestane (SHIP 1 inhibitor), SF1670 (PTEN inhibitor) and LY294002 (PI‐3K inhibitor). In M‐MOP cells (left panel) and primary microglia (right panel), PI(4,5)P 2 (A), PI(3,4)P 2 (B) and PIP 3 (C) were detected. All data show the mean ± SD of three independent experiments and were analysed by two‐way ANOVA (log‐transformed data were used in C) with Sidak’s multiple comparison tests performed. * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001 (blue: P522 and red: R522).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Transformation Assay

    ly294002  (Echelon Biosciences)


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    Echelon Biosciences ly294002
    A . Fura2 340/380 time traces from M-CSF-differentiated macrophages derived from conditionally-immortalised macrophage precursor cell lines (M-MØP) of Plcg2 P522 mice (P522, blue) and Plcg2 R522 mice (R522, red). One set of cell lines, generated from male mice, is shown. Cells were exposed to 5µg/ml anti-FcγRII/III along with 20µM EGTA and 2µM Ionomycin as indicated. B . Fura2 340/380 time traces from primary microglia derived from the cortex of Plcg2 R522 mice (blue: P522) and Plcg2 R522 mice (red: R522) with or without pre-exposure for 2 hours with Edelfosine <t>(10µM).</t> Cells were exposed to 5µg/ml anti-FcγRII/III along with EGTA and 2µM Ionomycin. Data in A and B shows the mean±SD of 3 independent experiments analysed by two-way ANOVA with Sidak post-hoc tests. PLCG2 R522 hiPSC-derived microglia show increased cytosolic Ca 2+ influx in comparison to controls following activation of PLCγ2 using anti-CD32. Cytoplasmic Ca2+ increase following activation of PLCγ2 using anti-CD32 Representative Ca 2+ traces ( C ) and graphical summary ( D ). 3 independent PLCG2 R522 CRISPR-engineered clones were examined. Data shown is mean±SD of 4 independent experiments and were analysed by one-way ANOVA with Tukey’s multiple comparison test (** = p<0.01; *** = p<0.001; **** = p< 0.0001).
    Ly294002, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "The Alzheimer’s disease protective P522R variant of PLCG2 , consistently enhances stimulus-dependent PLCγ2 activation, depleting substrate and altering cell function"

    Article Title: The Alzheimer’s disease protective P522R variant of PLCG2 , consistently enhances stimulus-dependent PLCγ2 activation, depleting substrate and altering cell function

    Journal: bioRxiv

    doi: 10.1101/2020.04.27.059600

    A . Fura2 340/380 time traces from M-CSF-differentiated macrophages derived from conditionally-immortalised macrophage precursor cell lines (M-MØP) of Plcg2 P522 mice (P522, blue) and Plcg2 R522 mice (R522, red). One set of cell lines, generated from male mice, is shown. Cells were exposed to 5µg/ml anti-FcγRII/III along with 20µM EGTA and 2µM Ionomycin as indicated. B . Fura2 340/380 time traces from primary microglia derived from the cortex of Plcg2 R522 mice (blue: P522) and Plcg2 R522 mice (red: R522) with or without pre-exposure for 2 hours with Edelfosine (10µM). Cells were exposed to 5µg/ml anti-FcγRII/III along with EGTA and 2µM Ionomycin. Data in A and B shows the mean±SD of 3 independent experiments analysed by two-way ANOVA with Sidak post-hoc tests. PLCG2 R522 hiPSC-derived microglia show increased cytosolic Ca 2+ influx in comparison to controls following activation of PLCγ2 using anti-CD32. Cytoplasmic Ca2+ increase following activation of PLCγ2 using anti-CD32 Representative Ca 2+ traces ( C ) and graphical summary ( D ). 3 independent PLCG2 R522 CRISPR-engineered clones were examined. Data shown is mean±SD of 4 independent experiments and were analysed by one-way ANOVA with Tukey’s multiple comparison test (** = p<0.01; *** = p<0.001; **** = p< 0.0001).
    Figure Legend Snippet: A . Fura2 340/380 time traces from M-CSF-differentiated macrophages derived from conditionally-immortalised macrophage precursor cell lines (M-MØP) of Plcg2 P522 mice (P522, blue) and Plcg2 R522 mice (R522, red). One set of cell lines, generated from male mice, is shown. Cells were exposed to 5µg/ml anti-FcγRII/III along with 20µM EGTA and 2µM Ionomycin as indicated. B . Fura2 340/380 time traces from primary microglia derived from the cortex of Plcg2 R522 mice (blue: P522) and Plcg2 R522 mice (red: R522) with or without pre-exposure for 2 hours with Edelfosine (10µM). Cells were exposed to 5µg/ml anti-FcγRII/III along with EGTA and 2µM Ionomycin. Data in A and B shows the mean±SD of 3 independent experiments analysed by two-way ANOVA with Sidak post-hoc tests. PLCG2 R522 hiPSC-derived microglia show increased cytosolic Ca 2+ influx in comparison to controls following activation of PLCγ2 using anti-CD32. Cytoplasmic Ca2+ increase following activation of PLCγ2 using anti-CD32 Representative Ca 2+ traces ( C ) and graphical summary ( D ). 3 independent PLCG2 R522 CRISPR-engineered clones were examined. Data shown is mean±SD of 4 independent experiments and were analysed by one-way ANOVA with Tukey’s multiple comparison test (** = p<0.01; *** = p<0.001; **** = p< 0.0001).

    Techniques Used: Derivative Assay, Generated, Activation Assay, CRISPR, Clone Assay

    A . M-MØP (blue: P522; red: R522) were loaded with Fluo-8 Ca 2+ indicator and examined for peak changes in fluoresce after exposure to 5ug/ml anti-FcγRII/III (2.4G2) with or without pre-exposure for 2 hours with Edelfosine (10µM) or U73122 (5µM) or Xestospogin C (5µM). B . Left panel shows LNA GapmeR percentage knockdown of Plcg2 gene expression in M-MOP cells within the P522 control (blue) and R522 (red) variant lines. Right panel shows Fura2 340/380 time traces from M-MØP cell lines (blue: P522; red: R522) which have undergone Plcg2 knockdown (square symbols) using an antisense LNA GapmeR. Cells were exposed to 5µg/ml anti-FcγRII/III and 2µM Ionomycin. C . Microglia from Plcg2 P522 (blue: P522) mice and Plcg2 R522 mice (red: R522) from neonate cortex (solid colour) or hippocampus (striped) were loaded with Fluo-8 Ca 2+ indicator. These cells were then examined for peak changes in fluoresce after exposure to 5µg/ml anti-FcγRII/III. These readings were taken with or without pre-exposure for 2 hours with Edelfosine (10µM) or U73122 (5µM). D Cortical microglia from Plcg2 P522 (blue: P522) mice and Plcg2 R522 mice (red: R522) from neonate cortex (solid colour) or hippocampus (striped) were loaded with Fluo-8 Ca 2+ indicator. These cells were then examined for peak changes in fluoresce after exposure to LPS (50ng/ml) or Aβ 1-42 oligomers (40µM). Data shown as the mean±SD of 3 independent experiments. Data in A, C and D were analysed by two-way ANOVA with Dunnett’s multiple Comparison test. See also Supplementary figure 3.
    Figure Legend Snippet: A . M-MØP (blue: P522; red: R522) were loaded with Fluo-8 Ca 2+ indicator and examined for peak changes in fluoresce after exposure to 5ug/ml anti-FcγRII/III (2.4G2) with or without pre-exposure for 2 hours with Edelfosine (10µM) or U73122 (5µM) or Xestospogin C (5µM). B . Left panel shows LNA GapmeR percentage knockdown of Plcg2 gene expression in M-MOP cells within the P522 control (blue) and R522 (red) variant lines. Right panel shows Fura2 340/380 time traces from M-MØP cell lines (blue: P522; red: R522) which have undergone Plcg2 knockdown (square symbols) using an antisense LNA GapmeR. Cells were exposed to 5µg/ml anti-FcγRII/III and 2µM Ionomycin. C . Microglia from Plcg2 P522 (blue: P522) mice and Plcg2 R522 mice (red: R522) from neonate cortex (solid colour) or hippocampus (striped) were loaded with Fluo-8 Ca 2+ indicator. These cells were then examined for peak changes in fluoresce after exposure to 5µg/ml anti-FcγRII/III. These readings were taken with or without pre-exposure for 2 hours with Edelfosine (10µM) or U73122 (5µM). D Cortical microglia from Plcg2 P522 (blue: P522) mice and Plcg2 R522 mice (red: R522) from neonate cortex (solid colour) or hippocampus (striped) were loaded with Fluo-8 Ca 2+ indicator. These cells were then examined for peak changes in fluoresce after exposure to LPS (50ng/ml) or Aβ 1-42 oligomers (40µM). Data shown as the mean±SD of 3 independent experiments. Data in A, C and D were analysed by two-way ANOVA with Dunnett’s multiple Comparison test. See also Supplementary figure 3.

    Techniques Used: Expressing, Variant Assay

    A mass ELISA was used to detect specific PIP species after exposure to anti-FcγRII/III (2.4G2) with or without 3-a-aminocholestane (SHIP 1 inhibitor), SF1670 (PTEN inhibitor) and LY294002 (PI-3K inhibitor). In M-MOP cells (left panel) and primary microglia (right panel) PI(4,5)P 2 (A) , PI(3,4)P 2 (B) and PIP 3 (C) were detected. All data shows the mean±SD of 3 independent experiments were analysed by 2 way ANOVA (log-transformed data was used in C) with Sidak’s multiple comparison tests performed *=p<0.05, **=p<0.01, ***=p<0.001, ****=p<0.0001. (blue: P522; red: R522)
    Figure Legend Snippet: A mass ELISA was used to detect specific PIP species after exposure to anti-FcγRII/III (2.4G2) with or without 3-a-aminocholestane (SHIP 1 inhibitor), SF1670 (PTEN inhibitor) and LY294002 (PI-3K inhibitor). In M-MOP cells (left panel) and primary microglia (right panel) PI(4,5)P 2 (A) , PI(3,4)P 2 (B) and PIP 3 (C) were detected. All data shows the mean±SD of 3 independent experiments were analysed by 2 way ANOVA (log-transformed data was used in C) with Sidak’s multiple comparison tests performed *=p<0.05, **=p<0.01, ***=p<0.001, ****=p<0.0001. (blue: P522; red: R522)

    Techniques Used: Enzyme-linked Immunosorbent Assay, Transformation Assay

    ly294002  (Echelon Biosciences)


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    Echelon Biosciences ly294002
    Triple pJMCS-LG3, pGNG1 and YCpLG-p110α YPH499 transformants were analyzed by flow cytometry as in in the presence of different doses of inhibitors as noted. The abscissae represent fold increase in green fluorescence of treated samples with the indicated concentrations of compounds <t>LY294002</t> (A) , 15e (B) , GDC-0326 (C) and ZSTK474 (D) relative to their correspondent DMSO alone control (normalized to 1). Three different clones were analyzed per experiment (n = 10,000 cells per clone). Data correspond to the average, and error bars represent SD. Asterisks (*) mark data points that are statistically significant respective to the data of the lowest concentration of compound tested in each series, according to Student's t-test (p < 0.05).
    Ly294002, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "A humanized yeast-based toolkit for monitoring phosphatidylinositol 3-kinase activity at both single cell and population levels"

    Article Title: A humanized yeast-based toolkit for monitoring phosphatidylinositol 3-kinase activity at both single cell and population levels

    Journal: Microbial Cell

    doi: 10.15698/mic2018.12.660

    Triple pJMCS-LG3, pGNG1 and YCpLG-p110α YPH499 transformants were analyzed by flow cytometry as in in the presence of different doses of inhibitors as noted. The abscissae represent fold increase in green fluorescence of treated samples with the indicated concentrations of compounds LY294002 (A) , 15e (B) , GDC-0326 (C) and ZSTK474 (D) relative to their correspondent DMSO alone control (normalized to 1). Three different clones were analyzed per experiment (n = 10,000 cells per clone). Data correspond to the average, and error bars represent SD. Asterisks (*) mark data points that are statistically significant respective to the data of the lowest concentration of compound tested in each series, according to Student's t-test (p < 0.05).
    Figure Legend Snippet: Triple pJMCS-LG3, pGNG1 and YCpLG-p110α YPH499 transformants were analyzed by flow cytometry as in in the presence of different doses of inhibitors as noted. The abscissae represent fold increase in green fluorescence of treated samples with the indicated concentrations of compounds LY294002 (A) , 15e (B) , GDC-0326 (C) and ZSTK474 (D) relative to their correspondent DMSO alone control (normalized to 1). Three different clones were analyzed per experiment (n = 10,000 cells per clone). Data correspond to the average, and error bars represent SD. Asterisks (*) mark data points that are statistically significant respective to the data of the lowest concentration of compound tested in each series, according to Student's t-test (p < 0.05).

    Techniques Used: Flow Cytometry, Fluorescence, Clone Assay, Concentration Assay

    ly294002  (Echelon Biosciences)


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    Echelon Biosciences ly294002
    Ly294002, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ly294002  (Echelon Biosciences)


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    Echelon Biosciences ly294002
    Ly294002, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Echelon Biosciences ly294002
    Standard plot of absorbance at 450 nm versus (A) log pmol PI(4,5)P2 and (C) log pmol PI3P. B Percentage change in PI(4,5)P2 content after BL irradiation in control (10 mM PIPES), neomycin (250 µM) and U73122 (25 µM) samples. D Percentage change in PI3P level after BL irradiation in control (10 mM PIPES), WM (10 µM) and <t>LY294002</t> (200 µM). The vertical bars represent SE of 3 independent experiments.
    Ly294002, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Echelon Biosciences inhibitors
    Standard plot of absorbance at 450 nm versus (A) log pmol PI(4,5)P2 and (C) log pmol PI3P. B Percentage change in PI(4,5)P2 content after BL irradiation in control (10 mM PIPES), neomycin (250 µM) and U73122 (25 µM) samples. D Percentage change in PI3P level after BL irradiation in control (10 mM PIPES), WM (10 µM) and <t>LY294002</t> (200 µM). The vertical bars represent SE of 3 independent experiments.
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    Standard plot of absorbance at 450 nm versus (A) log pmol PI(4,5)P2 and (C) log pmol PI3P. B Percentage change in PI(4,5)P2 content after BL irradiation in control (10 mM PIPES), neomycin (250 µM) and U73122 (25 µM) samples. D Percentage change in PI3P level after BL irradiation in control (10 mM PIPES), WM (10 µM) and LY294002 (200 µM). The vertical bars represent SE of 3 independent experiments.

    Journal: PLoS ONE

    Article Title: Phosphoinositides Play Differential Roles in Regulating Phototropin1- and Phototropin2-Mediated Chloroplast Movements in Arabidopsis

    doi: 10.1371/journal.pone.0055393

    Figure Lengend Snippet: Standard plot of absorbance at 450 nm versus (A) log pmol PI(4,5)P2 and (C) log pmol PI3P. B Percentage change in PI(4,5)P2 content after BL irradiation in control (10 mM PIPES), neomycin (250 µM) and U73122 (25 µM) samples. D Percentage change in PI3P level after BL irradiation in control (10 mM PIPES), WM (10 µM) and LY294002 (200 µM). The vertical bars represent SE of 3 independent experiments.

    Article Snippet: In a manner similar to PI(4,5)P2, the effects of WM and LY294002 on PI3K were also tested using the Echelon Bioscience mass Elisa kit (K3300).

    Techniques: Irradiation

    Leaves were infiltrated with ( A and B ) WM (10 µM) or ( C and D ) LY294002 (200 µM) and incubated for 1.5 h under darkness. Parameters of chloroplast responses: (A, C) amplitudes and (B, D) kinetics after treatment with inhibitors. Final readings are presented as mean ± SE (n = 10) from 2 independent experiments.

    Journal: PLoS ONE

    Article Title: Phosphoinositides Play Differential Roles in Regulating Phototropin1- and Phototropin2-Mediated Chloroplast Movements in Arabidopsis

    doi: 10.1371/journal.pone.0055393

    Figure Lengend Snippet: Leaves were infiltrated with ( A and B ) WM (10 µM) or ( C and D ) LY294002 (200 µM) and incubated for 1.5 h under darkness. Parameters of chloroplast responses: (A, C) amplitudes and (B, D) kinetics after treatment with inhibitors. Final readings are presented as mean ± SE (n = 10) from 2 independent experiments.

    Article Snippet: In a manner similar to PI(4,5)P2, the effects of WM and LY294002 on PI3K were also tested using the Echelon Bioscience mass Elisa kit (K3300).

    Techniques: Incubation

    A Leaf discs were illuminated with 100 µmol m −2 s −1 of BL for 5 s. B and C Leaf discs treated with (B) 10/25 µM U73122 or U73343 and (C) 10 µM WM, 50 µM WM or 200 µM LY294002 60 min before the measurements and then irradiated with 100 µmol m −2 s −1 BL for 5 s. The vertical bars represent SE of 20 independent experiments for each. The arrow shows the onset of BL pulse.

    Journal: PLoS ONE

    Article Title: Phosphoinositides Play Differential Roles in Regulating Phototropin1- and Phototropin2-Mediated Chloroplast Movements in Arabidopsis

    doi: 10.1371/journal.pone.0055393

    Figure Lengend Snippet: A Leaf discs were illuminated with 100 µmol m −2 s −1 of BL for 5 s. B and C Leaf discs treated with (B) 10/25 µM U73122 or U73343 and (C) 10 µM WM, 50 µM WM or 200 µM LY294002 60 min before the measurements and then irradiated with 100 µmol m −2 s −1 BL for 5 s. The vertical bars represent SE of 20 independent experiments for each. The arrow shows the onset of BL pulse.

    Article Snippet: In a manner similar to PI(4,5)P2, the effects of WM and LY294002 on PI3K were also tested using the Echelon Bioscience mass Elisa kit (K3300).

    Techniques: Irradiation