brp lpa  (Echelon Biosciences)


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    Echelon Biosciences brp lpa
    Brp Lpa, 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
    https://www.bioz.com/result/brp lpa/product/Echelon Biosciences
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    brp lpa - by Bioz Stars, 2023-06
    93/100 stars

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    brp lpa  (Echelon Biosciences)


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    Structured Review

    Echelon Biosciences brp lpa
    Brp Lpa, 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
    https://www.bioz.com/result/brp lpa/product/Echelon Biosciences
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    brp lpa - by Bioz Stars, 2023-06
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    α bromomethylene phosphonate lpa brp lpa  (Echelon Biosciences)


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    Echelon Biosciences α bromomethylene phosphonate lpa brp lpa
    (A) HUVEC or (B) bEnd.3 cells were treated with vehicle control (▪) or 5 µM <t>BrP-LPA</t> (▴) for 45 min prior to irradiation. (C) HUVEC were treated with 0.1% fatty acid free BSA (•), 10 µM LPA (○) or 10 µM LPA plus 5 µM BrP-LPA (▪) in serum free medium for 45 min prior to irradiation. The cells were then irradiated with 0, 2, 4 and 6 Gy and plated for clonogenic survival assay. After 2–3 wks, cells were stained with 1% methylene blue and colonies consisting of >50 cells were counted by microscopy. Surviving colonies were normalized for plating efficiency. Shown are average survival fractions and SEM from three experiments; * p<0.05. (D) Equal numbers of HUVEC were plated in 96 well plates and treated with carrier control 3% fatty acid free BSA, 10 µM LPA or 10 µM LPA with 5 µM BrP-LPA in serum free medium for 45 min prior to irradiation. After 96 h, the cell viability was determined using a colorimetric cell proliferation assay (Promega). Shown is the average absorbance at 490 nm with SEM from three experiments; * p<0.05.
    α Bromomethylene Phosphonate Lpa Brp Lpa, 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
    https://www.bioz.com/result/α bromomethylene phosphonate lpa brp lpa/product/Echelon Biosciences
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    α bromomethylene phosphonate lpa brp lpa - by Bioz Stars, 2023-06
    93/100 stars

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    1) Product Images from "Autotaxin and LPA Receptors Represent Potential Molecular Targets for the Radiosensitization of Murine Glioma through Effects on Tumor Vasculature"

    Article Title: Autotaxin and LPA Receptors Represent Potential Molecular Targets for the Radiosensitization of Murine Glioma through Effects on Tumor Vasculature

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0022182

    (A) HUVEC or (B) bEnd.3 cells were treated with vehicle control (▪) or 5 µM BrP-LPA (▴) for 45 min prior to irradiation. (C) HUVEC were treated with 0.1% fatty acid free BSA (•), 10 µM LPA (○) or 10 µM LPA plus 5 µM BrP-LPA (▪) in serum free medium for 45 min prior to irradiation. The cells were then irradiated with 0, 2, 4 and 6 Gy and plated for clonogenic survival assay. After 2–3 wks, cells were stained with 1% methylene blue and colonies consisting of >50 cells were counted by microscopy. Surviving colonies were normalized for plating efficiency. Shown are average survival fractions and SEM from three experiments; * p<0.05. (D) Equal numbers of HUVEC were plated in 96 well plates and treated with carrier control 3% fatty acid free BSA, 10 µM LPA or 10 µM LPA with 5 µM BrP-LPA in serum free medium for 45 min prior to irradiation. After 96 h, the cell viability was determined using a colorimetric cell proliferation assay (Promega). Shown is the average absorbance at 490 nm with SEM from three experiments; * p<0.05.
    Figure Legend Snippet: (A) HUVEC or (B) bEnd.3 cells were treated with vehicle control (▪) or 5 µM BrP-LPA (▴) for 45 min prior to irradiation. (C) HUVEC were treated with 0.1% fatty acid free BSA (•), 10 µM LPA (○) or 10 µM LPA plus 5 µM BrP-LPA (▪) in serum free medium for 45 min prior to irradiation. The cells were then irradiated with 0, 2, 4 and 6 Gy and plated for clonogenic survival assay. After 2–3 wks, cells were stained with 1% methylene blue and colonies consisting of >50 cells were counted by microscopy. Surviving colonies were normalized for plating efficiency. Shown are average survival fractions and SEM from three experiments; * p<0.05. (D) Equal numbers of HUVEC were plated in 96 well plates and treated with carrier control 3% fatty acid free BSA, 10 µM LPA or 10 µM LPA with 5 µM BrP-LPA in serum free medium for 45 min prior to irradiation. After 96 h, the cell viability was determined using a colorimetric cell proliferation assay (Promega). Shown is the average absorbance at 490 nm with SEM from three experiments; * p<0.05.

    Techniques Used: Irradiation, Clonogenic Cell Survival Assay, Staining, Microscopy, Proliferation Assay

    (A, B) HUVEC or (C) bEnd.3 cells were plated on matrigel-coated 96 well plates and treated with vehicle control or 5 µM BrP-LPA for 45 min prior to irradiation with 3 Gy. (A) Shown are the representative photomicrographs of tubule formation taken 16 h after irradiation. (B) Tubule formation was quantified as number of tubules per high power field (HPF). Shown are bar graphs of mean number of tubules per HPF relative to control and SEM from three experiments; * p<0.05.
    Figure Legend Snippet: (A, B) HUVEC or (C) bEnd.3 cells were plated on matrigel-coated 96 well plates and treated with vehicle control or 5 µM BrP-LPA for 45 min prior to irradiation with 3 Gy. (A) Shown are the representative photomicrographs of tubule formation taken 16 h after irradiation. (B) Tubule formation was quantified as number of tubules per high power field (HPF). Shown are bar graphs of mean number of tubules per HPF relative to control and SEM from three experiments; * p<0.05.

    Techniques Used: Irradiation

    (A) bEnd.3 or (B) HUVEC were plated on 60 mm plates and allowed to grow to 70% confluency. The semi-confluent cell layer was scraped using a sterile pipette tip to create a scratch devoid of cells. The remaining cells were treated with vehicle control or 5 µM BrP-LPA for 45 min prior to irradiation with 3 Gy. Migration was observed at 36 h. Cells were fixed with ethanol and stained with 1% methylene blue. Migrated cells were counted and normalized to surrounding cell density per HPF. Shown are representative photomicrographs and bar graphs representing the mean percentages of migrating cells relative to corresponding controls with SEM from three experiments, * p<0.05.
    Figure Legend Snippet: (A) bEnd.3 or (B) HUVEC were plated on 60 mm plates and allowed to grow to 70% confluency. The semi-confluent cell layer was scraped using a sterile pipette tip to create a scratch devoid of cells. The remaining cells were treated with vehicle control or 5 µM BrP-LPA for 45 min prior to irradiation with 3 Gy. Migration was observed at 36 h. Cells were fixed with ethanol and stained with 1% methylene blue. Migrated cells were counted and normalized to surrounding cell density per HPF. Shown are representative photomicrographs and bar graphs representing the mean percentages of migrating cells relative to corresponding controls with SEM from three experiments, * p<0.05.

    Techniques Used: Transferring, Irradiation, Migration, Staining

    (A, B) Mouse glioma GL261 cells were plated on 60 mm plates and allowed to grow to 70% confluency. Plates were scraped with a pipette tip to create a scratch devoid of cells and treated with vehicle control or 5 µM BrP-LPA for 45 min before irradiation with 3 Gy. After 24 h, cells were fixed and stained with methylene blue. Migrated cells were counted and normalized to surrounding cell density per HPF. Shown are representative photomicrographs (A) and a bar graph (B) representing the mean percentages of migrating cells relative to corresponding controls with SEM from three experiments; * p<0.05. (C) For clonogenic survival assay, GL261 cells were plated and allowed to attach. After 6 h, cells were treated vehicle control or with 5 µM BrP-LPA for 45 min and irradiated with 0, 2, 4, and 6 Gy. After 10 days, surviving colonies (>50 cells) were counted and normalized for plating efficiency. Shown is the clonogenic survival curve and mean surviving fractions and SEM from three experiments; * p<0.05.
    Figure Legend Snippet: (A, B) Mouse glioma GL261 cells were plated on 60 mm plates and allowed to grow to 70% confluency. Plates were scraped with a pipette tip to create a scratch devoid of cells and treated with vehicle control or 5 µM BrP-LPA for 45 min before irradiation with 3 Gy. After 24 h, cells were fixed and stained with methylene blue. Migrated cells were counted and normalized to surrounding cell density per HPF. Shown are representative photomicrographs (A) and a bar graph (B) representing the mean percentages of migrating cells relative to corresponding controls with SEM from three experiments; * p<0.05. (C) For clonogenic survival assay, GL261 cells were plated and allowed to attach. After 6 h, cells were treated vehicle control or with 5 µM BrP-LPA for 45 min and irradiated with 0, 2, 4, and 6 Gy. After 10 days, surviving colonies (>50 cells) were counted and normalized for plating efficiency. Shown is the clonogenic survival curve and mean surviving fractions and SEM from three experiments; * p<0.05.

    Techniques Used: Transferring, Irradiation, Staining, Clonogenic Cell Survival Assay

    bEnd.3 and GL261 cells were grown in co-culture for 24 h. Cells were treated with vehicle or 5 µM BrP-LPA control for 45 min before treatment with 3 Gy. Cells were lysed at 5 min after irradiation. Shown are immunoblot analyses using specific antibodies to phospho-Akt Thr308/Ser473 , total Akt, and actin. Numbers represent the ratios of phospho-Akt to total Akt protein normalized to actin (fold change relative to control).
    Figure Legend Snippet: bEnd.3 and GL261 cells were grown in co-culture for 24 h. Cells were treated with vehicle or 5 µM BrP-LPA control for 45 min before treatment with 3 Gy. Cells were lysed at 5 min after irradiation. Shown are immunoblot analyses using specific antibodies to phospho-Akt Thr308/Ser473 , total Akt, and actin. Numbers represent the ratios of phospho-Akt to total Akt protein normalized to actin (fold change relative to control).

    Techniques Used: Co-Culture Assay, Irradiation, Western Blot

    GL261 cells were injected into the hind limbs of nude mice. Tumors were irradiated with 3 Gy for 5 consecutive days for a total of 15 Gy. Mice were treated with 3 mg/kg BrP-LPA or vehicle control for 45 min prior to irradiation on days 1, 3, and 5. (A) Shown are mean tumor volumes with SEM from each treatment group of 5 mice. (B) Tumor growth delay was calculated as the number of days for tumors to reach a 6-fold volume increase compared to control. Shown is a bar graph representing the mean tumor growth delay with SEM from each treatment group of 5 mice; * p<0.05.
    Figure Legend Snippet: GL261 cells were injected into the hind limbs of nude mice. Tumors were irradiated with 3 Gy for 5 consecutive days for a total of 15 Gy. Mice were treated with 3 mg/kg BrP-LPA or vehicle control for 45 min prior to irradiation on days 1, 3, and 5. (A) Shown are mean tumor volumes with SEM from each treatment group of 5 mice. (B) Tumor growth delay was calculated as the number of days for tumors to reach a 6-fold volume increase compared to control. Shown is a bar graph representing the mean tumor growth delay with SEM from each treatment group of 5 mice; * p<0.05.

    Techniques Used: Injection, Irradiation

    brp lpa  (Echelon Biosciences)


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    Echelon Biosciences brp lpa
    A–C. Steady-state hydrolysis of LPC (16:0, 18:0, 18:1 respectively) by recombinant ATX as measured with the TOOS activity assay. D–E. ATX activity inhibition dose-response curves in the presence of various <t>BrP-LPA</t> concentrations as measured with the TOOS assay using as substrates 50 μΜ of 16:0 (D) and 18:0 LPC (E). F–G . Cornish-Bowden (F) and Lineweaver-Burk (G) plots show that BrP-LPA is a competitive ATX inhibitor. H. The kinetic parameters (k m , V max ) obtained by the incubation of ATX at various concentrations of 16:0 LPC (100–400 μΜ) in the absence or presence of BrPLPA (0–10 μΜ). All presented values are the means (±std) of two independent experiments.
    Brp Lpa, 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
    https://www.bioz.com/result/brp lpa/product/Echelon Biosciences
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    brp lpa - by Bioz Stars, 2023-06
    93/100 stars

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    1) Product Images from "A Metabolically-Stabilized Phosphonate Analog of Lysophosphatidic Acid Attenuates Collagen-Induced Arthritis"

    Article Title: A Metabolically-Stabilized Phosphonate Analog of Lysophosphatidic Acid Attenuates Collagen-Induced Arthritis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070941

    A–C. Steady-state hydrolysis of LPC (16:0, 18:0, 18:1 respectively) by recombinant ATX as measured with the TOOS activity assay. D–E. ATX activity inhibition dose-response curves in the presence of various BrP-LPA concentrations as measured with the TOOS assay using as substrates 50 μΜ of 16:0 (D) and 18:0 LPC (E). F–G . Cornish-Bowden (F) and Lineweaver-Burk (G) plots show that BrP-LPA is a competitive ATX inhibitor. H. The kinetic parameters (k m , V max ) obtained by the incubation of ATX at various concentrations of 16:0 LPC (100–400 μΜ) in the absence or presence of BrPLPA (0–10 μΜ). All presented values are the means (±std) of two independent experiments.
    Figure Legend Snippet: A–C. Steady-state hydrolysis of LPC (16:0, 18:0, 18:1 respectively) by recombinant ATX as measured with the TOOS activity assay. D–E. ATX activity inhibition dose-response curves in the presence of various BrP-LPA concentrations as measured with the TOOS assay using as substrates 50 μΜ of 16:0 (D) and 18:0 LPC (E). F–G . Cornish-Bowden (F) and Lineweaver-Burk (G) plots show that BrP-LPA is a competitive ATX inhibitor. H. The kinetic parameters (k m , V max ) obtained by the incubation of ATX at various concentrations of 16:0 LPC (100–400 μΜ) in the absence or presence of BrPLPA (0–10 μΜ). All presented values are the means (±std) of two independent experiments.

    Techniques Used: Recombinant, Activity Assay, Inhibition, Incubation

    Kinetic parameters of in vitro LPC hydrolysis by recombinant ATX and inhibition constants of  BrP-LPA.
    Figure Legend Snippet: Kinetic parameters of in vitro LPC hydrolysis by recombinant ATX and inhibition constants of BrP-LPA.

    Techniques Used: In Vitro, Recombinant, Inhibition

    A. Effect of BrP-LPA on plasma ATX/lysoPLD hydrolysis of exogenous added 50 μΜ LPC (14:0, 16:0, 18:0). B. Correlation of recovered BrP-LPA in whole blood with the added BrP-LPA. C. Inhibition of endogenous ATX/lysoPLD activity in whole blood ex vivo after the addition of increasing BrP-LPA concentrations (0–10 μΜ). ATX activity was measured in the presence of 1 mM LPC with the TOOS assay. D. LPC levels and E. LPA levels measured in whole blood ex vivo in the absence/presence of different BrP-LPA concentrations (0.03–10 μΜ). F. Per cent residual levels of the indicated LPA species in the presence of increasing amounts of BrP-LPA. Solid lines, best fits of averaged data points. * indicates significant (p<0.05), *** indicates significant (p<0.001) decrease relative to control group.
    Figure Legend Snippet: A. Effect of BrP-LPA on plasma ATX/lysoPLD hydrolysis of exogenous added 50 μΜ LPC (14:0, 16:0, 18:0). B. Correlation of recovered BrP-LPA in whole blood with the added BrP-LPA. C. Inhibition of endogenous ATX/lysoPLD activity in whole blood ex vivo after the addition of increasing BrP-LPA concentrations (0–10 μΜ). ATX activity was measured in the presence of 1 mM LPC with the TOOS assay. D. LPC levels and E. LPA levels measured in whole blood ex vivo in the absence/presence of different BrP-LPA concentrations (0.03–10 μΜ). F. Per cent residual levels of the indicated LPA species in the presence of increasing amounts of BrP-LPA. Solid lines, best fits of averaged data points. * indicates significant (p<0.05), *** indicates significant (p<0.001) decrease relative to control group.

    Techniques Used: Inhibition, Activity Assay, Ex Vivo

    Inhibition constants as measured by dose-response curves obtained by residual percent activity of plasma ATX/LysoPLD (TOOS activity assay) after the addition of increasing amounts of  BrP-LPA  (0.01–100 μM) in the presence of different exogenous LPC species.
    Figure Legend Snippet: Inhibition constants as measured by dose-response curves obtained by residual percent activity of plasma ATX/LysoPLD (TOOS activity assay) after the addition of increasing amounts of BrP-LPA (0.01–100 μM) in the presence of different exogenous LPC species.

    Techniques Used: Inhibition, Activity Assay

    A. Plasma BrP-LPA pharmacokinetic profile following i.p administration of 10 mg/Kg BrP-LPA in female mice. B. Plasma per cent (%) residual ATX activity measured in the presence of 1 mM LPC with the TOOS assay and C . Plasma % residual total LPA levels at different time points after i.p. administration of BrP-LPA. D. Plasma concentration of different LPA and E. LPC species at different time points following BrP-LPA i.p administration. The time point 0 refers to the vehicle control. The presented values are the means (±std) of two independent experiments. *(p<0.05) and **(p<0.01) indicate a significant decrease relative to control group.
    Figure Legend Snippet: A. Plasma BrP-LPA pharmacokinetic profile following i.p administration of 10 mg/Kg BrP-LPA in female mice. B. Plasma per cent (%) residual ATX activity measured in the presence of 1 mM LPC with the TOOS assay and C . Plasma % residual total LPA levels at different time points after i.p. administration of BrP-LPA. D. Plasma concentration of different LPA and E. LPC species at different time points following BrP-LPA i.p administration. The time point 0 refers to the vehicle control. The presented values are the means (±std) of two independent experiments. *(p<0.05) and **(p<0.01) indicate a significant decrease relative to control group.

    Techniques Used: Activity Assay, Concentration Assay

    A. Schematic representation of immunization and drug administration schemes. DBA/1 mice subjected to CIA were treated intraperitoneally with vehicle, BrP-LPA (10 mg/kg), or dexamethasone (Dex, 3 mg/kg) twice a week. B–C. Time dependent clinical scores (± SEM) after primary immunization with CII. Data are pooled from two different experiments (n = 10). D. Representative sections of the forelimb and hindlimb joints of vehicle-treated and BrP-LPA-treated mice, stained with H&E. E. Quantification of disease severity in vehicle-treated and BrP-LPA-treated mice. Joint sections were assessed histologically in a blinded manner by three independent examiners. Data are shown as mean joint values per mouse ± SD. F. Humoral response to collagen in vehicle-treated or BrP-LPA-treated mice. Levels of CII-specific Abs were determined by ELISA in mouse sera. Results show the mean ± SEM values. *(p<0.05) and **(p<0.01) indicate a significant decrease relative to control group.
    Figure Legend Snippet: A. Schematic representation of immunization and drug administration schemes. DBA/1 mice subjected to CIA were treated intraperitoneally with vehicle, BrP-LPA (10 mg/kg), or dexamethasone (Dex, 3 mg/kg) twice a week. B–C. Time dependent clinical scores (± SEM) after primary immunization with CII. Data are pooled from two different experiments (n = 10). D. Representative sections of the forelimb and hindlimb joints of vehicle-treated and BrP-LPA-treated mice, stained with H&E. E. Quantification of disease severity in vehicle-treated and BrP-LPA-treated mice. Joint sections were assessed histologically in a blinded manner by three independent examiners. Data are shown as mean joint values per mouse ± SD. F. Humoral response to collagen in vehicle-treated or BrP-LPA-treated mice. Levels of CII-specific Abs were determined by ELISA in mouse sera. Results show the mean ± SEM values. *(p<0.05) and **(p<0.01) indicate a significant decrease relative to control group.

    Techniques Used: Staining, Enzyme-linked Immunosorbent Assay

    brp lpa  (Echelon Biosciences)


    Bioz Verified Symbol Echelon Biosciences is a verified supplier
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    Echelon Biosciences brp lpa
    A–C. Steady-state hydrolysis of LPC (16:0, 18:0, 18:1 respectively) by recombinant ATX as measured with the TOOS activity assay. D–E. ATX activity inhibition dose-response curves in the presence of various <t>BrP-LPA</t> concentrations as measured with the TOOS assay using as substrates 50 μΜ of 16:0 (D) and 18:0 LPC (E). F–G . Cornish-Bowden (F) and Lineweaver-Burk (G) plots show that BrP-LPA is a competitive ATX inhibitor. H. The kinetic parameters (k m , V max ) obtained by the incubation of ATX at various concentrations of 16:0 LPC (100–400 μΜ) in the absence or presence of BrPLPA (0–10 μΜ). All presented values are the means (±std) of two independent experiments.
    Brp Lpa, 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
    https://www.bioz.com/result/brp lpa/product/Echelon Biosciences
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    brp lpa - by Bioz Stars, 2023-06
    93/100 stars

    Images

    1) Product Images from "A Metabolically-Stabilized Phosphonate Analog of Lysophosphatidic Acid Attenuates Collagen-Induced Arthritis"

    Article Title: A Metabolically-Stabilized Phosphonate Analog of Lysophosphatidic Acid Attenuates Collagen-Induced Arthritis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070941

    A–C. Steady-state hydrolysis of LPC (16:0, 18:0, 18:1 respectively) by recombinant ATX as measured with the TOOS activity assay. D–E. ATX activity inhibition dose-response curves in the presence of various BrP-LPA concentrations as measured with the TOOS assay using as substrates 50 μΜ of 16:0 (D) and 18:0 LPC (E). F–G . Cornish-Bowden (F) and Lineweaver-Burk (G) plots show that BrP-LPA is a competitive ATX inhibitor. H. The kinetic parameters (k m , V max ) obtained by the incubation of ATX at various concentrations of 16:0 LPC (100–400 μΜ) in the absence or presence of BrPLPA (0–10 μΜ). All presented values are the means (±std) of two independent experiments.
    Figure Legend Snippet: A–C. Steady-state hydrolysis of LPC (16:0, 18:0, 18:1 respectively) by recombinant ATX as measured with the TOOS activity assay. D–E. ATX activity inhibition dose-response curves in the presence of various BrP-LPA concentrations as measured with the TOOS assay using as substrates 50 μΜ of 16:0 (D) and 18:0 LPC (E). F–G . Cornish-Bowden (F) and Lineweaver-Burk (G) plots show that BrP-LPA is a competitive ATX inhibitor. H. The kinetic parameters (k m , V max ) obtained by the incubation of ATX at various concentrations of 16:0 LPC (100–400 μΜ) in the absence or presence of BrPLPA (0–10 μΜ). All presented values are the means (±std) of two independent experiments.

    Techniques Used: Recombinant, Activity Assay, Inhibition, Incubation

    Kinetic parameters of in vitro LPC hydrolysis by recombinant ATX and inhibition constants of  BrP-LPA.
    Figure Legend Snippet: Kinetic parameters of in vitro LPC hydrolysis by recombinant ATX and inhibition constants of BrP-LPA.

    Techniques Used: In Vitro, Recombinant, Inhibition

    A. Effect of BrP-LPA on plasma ATX/lysoPLD hydrolysis of exogenous added 50 μΜ LPC (14:0, 16:0, 18:0). B. Correlation of recovered BrP-LPA in whole blood with the added BrP-LPA. C. Inhibition of endogenous ATX/lysoPLD activity in whole blood ex vivo after the addition of increasing BrP-LPA concentrations (0–10 μΜ). ATX activity was measured in the presence of 1 mM LPC with the TOOS assay. D. LPC levels and E. LPA levels measured in whole blood ex vivo in the absence/presence of different BrP-LPA concentrations (0.03–10 μΜ). F. Per cent residual levels of the indicated LPA species in the presence of increasing amounts of BrP-LPA. Solid lines, best fits of averaged data points. * indicates significant (p<0.05), *** indicates significant (p<0.001) decrease relative to control group.
    Figure Legend Snippet: A. Effect of BrP-LPA on plasma ATX/lysoPLD hydrolysis of exogenous added 50 μΜ LPC (14:0, 16:0, 18:0). B. Correlation of recovered BrP-LPA in whole blood with the added BrP-LPA. C. Inhibition of endogenous ATX/lysoPLD activity in whole blood ex vivo after the addition of increasing BrP-LPA concentrations (0–10 μΜ). ATX activity was measured in the presence of 1 mM LPC with the TOOS assay. D. LPC levels and E. LPA levels measured in whole blood ex vivo in the absence/presence of different BrP-LPA concentrations (0.03–10 μΜ). F. Per cent residual levels of the indicated LPA species in the presence of increasing amounts of BrP-LPA. Solid lines, best fits of averaged data points. * indicates significant (p<0.05), *** indicates significant (p<0.001) decrease relative to control group.

    Techniques Used: Inhibition, Activity Assay, Ex Vivo

    Inhibition constants as measured by dose-response curves obtained by residual percent activity of plasma ATX/LysoPLD (TOOS activity assay) after the addition of increasing amounts of  BrP-LPA  (0.01–100 μM) in the presence of different exogenous LPC species.
    Figure Legend Snippet: Inhibition constants as measured by dose-response curves obtained by residual percent activity of plasma ATX/LysoPLD (TOOS activity assay) after the addition of increasing amounts of BrP-LPA (0.01–100 μM) in the presence of different exogenous LPC species.

    Techniques Used: Inhibition, Activity Assay

    A. Plasma BrP-LPA pharmacokinetic profile following i.p administration of 10 mg/Kg BrP-LPA in female mice. B. Plasma per cent (%) residual ATX activity measured in the presence of 1 mM LPC with the TOOS assay and C . Plasma % residual total LPA levels at different time points after i.p. administration of BrP-LPA. D. Plasma concentration of different LPA and E. LPC species at different time points following BrP-LPA i.p administration. The time point 0 refers to the vehicle control. The presented values are the means (±std) of two independent experiments. *(p<0.05) and **(p<0.01) indicate a significant decrease relative to control group.
    Figure Legend Snippet: A. Plasma BrP-LPA pharmacokinetic profile following i.p administration of 10 mg/Kg BrP-LPA in female mice. B. Plasma per cent (%) residual ATX activity measured in the presence of 1 mM LPC with the TOOS assay and C . Plasma % residual total LPA levels at different time points after i.p. administration of BrP-LPA. D. Plasma concentration of different LPA and E. LPC species at different time points following BrP-LPA i.p administration. The time point 0 refers to the vehicle control. The presented values are the means (±std) of two independent experiments. *(p<0.05) and **(p<0.01) indicate a significant decrease relative to control group.

    Techniques Used: Activity Assay, Concentration Assay

    A. Schematic representation of immunization and drug administration schemes. DBA/1 mice subjected to CIA were treated intraperitoneally with vehicle, BrP-LPA (10 mg/kg), or dexamethasone (Dex, 3 mg/kg) twice a week. B–C. Time dependent clinical scores (± SEM) after primary immunization with CII. Data are pooled from two different experiments (n = 10). D. Representative sections of the forelimb and hindlimb joints of vehicle-treated and BrP-LPA-treated mice, stained with H&E. E. Quantification of disease severity in vehicle-treated and BrP-LPA-treated mice. Joint sections were assessed histologically in a blinded manner by three independent examiners. Data are shown as mean joint values per mouse ± SD. F. Humoral response to collagen in vehicle-treated or BrP-LPA-treated mice. Levels of CII-specific Abs were determined by ELISA in mouse sera. Results show the mean ± SEM values. *(p<0.05) and **(p<0.01) indicate a significant decrease relative to control group.
    Figure Legend Snippet: A. Schematic representation of immunization and drug administration schemes. DBA/1 mice subjected to CIA were treated intraperitoneally with vehicle, BrP-LPA (10 mg/kg), or dexamethasone (Dex, 3 mg/kg) twice a week. B–C. Time dependent clinical scores (± SEM) after primary immunization with CII. Data are pooled from two different experiments (n = 10). D. Representative sections of the forelimb and hindlimb joints of vehicle-treated and BrP-LPA-treated mice, stained with H&E. E. Quantification of disease severity in vehicle-treated and BrP-LPA-treated mice. Joint sections were assessed histologically in a blinded manner by three independent examiners. Data are shown as mean joint values per mouse ± SD. F. Humoral response to collagen in vehicle-treated or BrP-LPA-treated mice. Levels of CII-specific Abs were determined by ELISA in mouse sera. Results show the mean ± SEM values. *(p<0.05) and **(p<0.01) indicate a significant decrease relative to control group.

    Techniques Used: Staining, Enzyme-linked Immunosorbent Assay

    brp lpa  (Echelon Biosciences)


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    Echelon Biosciences brp lpa
    Summary of stimulators and inhibitors tested
    Brp Lpa, 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 "Development of a selective fluorescence-based enzyme assay for glycerophosphodiesterase family members GDE4 and GDE7"

    Article Title: Development of a selective fluorescence-based enzyme assay for glycerophosphodiesterase family members GDE4 and GDE7

    Journal: Journal of Lipid Research

    doi: 10.1016/j.jlr.2021.100141

    Summary of stimulators and inhibitors tested
    Figure Legend Snippet: Summary of stimulators and inhibitors tested

    Techniques Used: Inhibition

    Effects of natural substrates, LPC analogs, and ATX inhibitors on FS-3-degrading enzyme activity of GDE4 and GDE7. The membrane fractions from the GDE4- (left graphs) or GDE7-expressing (right graphs) HEK293T cells were incubated with 5 μM FS-3 in the presence of 2 mM MgCl 2 (for GDE4) or 2 mM CaCl 2 (for GDE7). The amounts of protein in the membrane fractions were 5 μg (left graphs) and 1 μg (right graphs). The effects of LPC (5% ethanol final concentration, A), LPE (5% ethanol, B), edelfosine (C), S32826 (0.1–0.2% DMSO, D), BrP-LPA (0.1–0.2% DMSO, E), and 3-ccPA (10% methanol, F) were examined at the indicated concentrations, and FS-3-degrading activity is shown (mean ± SD, n = 3 or 4). One-way ANOVA with post hoc Dunnett test was conducted. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 (vs. vehicle).
    Figure Legend Snippet: Effects of natural substrates, LPC analogs, and ATX inhibitors on FS-3-degrading enzyme activity of GDE4 and GDE7. The membrane fractions from the GDE4- (left graphs) or GDE7-expressing (right graphs) HEK293T cells were incubated with 5 μM FS-3 in the presence of 2 mM MgCl 2 (for GDE4) or 2 mM CaCl 2 (for GDE7). The amounts of protein in the membrane fractions were 5 μg (left graphs) and 1 μg (right graphs). The effects of LPC (5% ethanol final concentration, A), LPE (5% ethanol, B), edelfosine (C), S32826 (0.1–0.2% DMSO, D), BrP-LPA (0.1–0.2% DMSO, E), and 3-ccPA (10% methanol, F) were examined at the indicated concentrations, and FS-3-degrading activity is shown (mean ± SD, n = 3 or 4). One-way ANOVA with post hoc Dunnett test was conducted. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 (vs. vehicle).

    Techniques Used: Activity Assay, Expressing, Incubation, Concentration Assay

    brp lpa  (Echelon Biosciences)


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    Echelon Biosciences brp lpa
    Brp Lpa, 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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    bromo  (Echelon Biosciences)


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    Echelon Biosciences bromo
    Bromo, 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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    brp lpa  (Echelon Biosciences)


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    Echelon Biosciences brp lpa
    Inhibition of LPA5 suppresses the <t>LPA-induced</t> pro-inflammatory phenotype in BV-2 and primary murine microglia. a Serum-starved BV-2 cells were treated with LPA in the absence or presence of <t>BrP-LPA</t> (5 μM; upper panel) or TCLPA5 (5 μM; lower panel) added 2 h prior to LPA addition. COX-2 and Arg-1 response was monitored using Western blotting. One representative plot for each protein and the densitometric analysis (mean + SD) from four independent experiments is presented. (** p < 0.01; *** p < 0.001; # p < 0.05, inhibitor compared to LPA-treated cells; one-way ANOVA with Bonferroni correction). b PMM were incubated in the presence of vehicle (DMSO; 'untreated'), LPA (1 μM), vehicle (DMSO) plus LPA (1 µM), or TCLPA5 (5 μM in DMSO; added 2 h prior to LPA addition) plus LPA (1 µM) for 24 h. Cells were stained for iNOS, COX-2, Arg-1, or RELMα and visualized using confocal microscopy. Fluorescence intensity was quantitated with ImageJ. At least 50 cells out of 3 different areas per chamber were measured. Results (three independent experiments) are presented as mean + SD (* p < 0.05; *** p < 0.001; # p < 0.05 inhibitor compared to LPA-treated cells; one-way ANOVA with Bonferroni correction)
    Brp Lpa, 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 "1-Oleyl-lysophosphatidic acid (LPA) promotes polarization of BV-2 and primary murine microglia towards an M1-like phenotype"

    Article Title: 1-Oleyl-lysophosphatidic acid (LPA) promotes polarization of BV-2 and primary murine microglia towards an M1-like phenotype

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-016-0701-9

    Inhibition of LPA5 suppresses the LPA-induced pro-inflammatory phenotype in BV-2 and primary murine microglia. a Serum-starved BV-2 cells were treated with LPA in the absence or presence of BrP-LPA (5 μM; upper panel) or TCLPA5 (5 μM; lower panel) added 2 h prior to LPA addition. COX-2 and Arg-1 response was monitored using Western blotting. One representative plot for each protein and the densitometric analysis (mean + SD) from four independent experiments is presented. (** p < 0.01; *** p < 0.001; # p < 0.05, inhibitor compared to LPA-treated cells; one-way ANOVA with Bonferroni correction). b PMM were incubated in the presence of vehicle (DMSO; 'untreated'), LPA (1 μM), vehicle (DMSO) plus LPA (1 µM), or TCLPA5 (5 μM in DMSO; added 2 h prior to LPA addition) plus LPA (1 µM) for 24 h. Cells were stained for iNOS, COX-2, Arg-1, or RELMα and visualized using confocal microscopy. Fluorescence intensity was quantitated with ImageJ. At least 50 cells out of 3 different areas per chamber were measured. Results (three independent experiments) are presented as mean + SD (* p < 0.05; *** p < 0.001; # p < 0.05 inhibitor compared to LPA-treated cells; one-way ANOVA with Bonferroni correction)
    Figure Legend Snippet: Inhibition of LPA5 suppresses the LPA-induced pro-inflammatory phenotype in BV-2 and primary murine microglia. a Serum-starved BV-2 cells were treated with LPA in the absence or presence of BrP-LPA (5 μM; upper panel) or TCLPA5 (5 μM; lower panel) added 2 h prior to LPA addition. COX-2 and Arg-1 response was monitored using Western blotting. One representative plot for each protein and the densitometric analysis (mean + SD) from four independent experiments is presented. (** p < 0.01; *** p < 0.001; # p < 0.05, inhibitor compared to LPA-treated cells; one-way ANOVA with Bonferroni correction). b PMM were incubated in the presence of vehicle (DMSO; 'untreated'), LPA (1 μM), vehicle (DMSO) plus LPA (1 µM), or TCLPA5 (5 μM in DMSO; added 2 h prior to LPA addition) plus LPA (1 µM) for 24 h. Cells were stained for iNOS, COX-2, Arg-1, or RELMα and visualized using confocal microscopy. Fluorescence intensity was quantitated with ImageJ. At least 50 cells out of 3 different areas per chamber were measured. Results (three independent experiments) are presented as mean + SD (* p < 0.05; *** p < 0.001; # p < 0.05 inhibitor compared to LPA-treated cells; one-way ANOVA with Bonferroni correction)

    Techniques Used: Inhibition, Western Blot, Incubation, Staining, Confocal Microscopy, Fluorescence

    LPA receptor antagonists attenuate M1 surface marker expression in BV-2 cells. Serum-starved (o/n) cells were cultivated in the presence of vehicle, LPA (1 μM), or LPA plus a BrP-LPA (5 μM) and b TCLPA5 (5 μM) for the indicated times. Inhibitors were added 2 h prior LPA addition. Cells were stained with PE-conjugated anti-CD40, APC-conjugated anti-CD86, or PE-conjugated anti-CD206 antibodies and analyzed using a Guava easyCyte 8 Millipore flow cytometer. Results from four individual experiments in triplicate are shown as mean values + SD. (* p < 0.05; ** p < 0.01; *** p < 0.001 compared to untreated or DMSO-treated cells; # p < 0.05; ## p < 0.01; ### p < 0.001 inhibitor compared to LPA-treated cells; one-way ANOVA with Bonferroni correction)
    Figure Legend Snippet: LPA receptor antagonists attenuate M1 surface marker expression in BV-2 cells. Serum-starved (o/n) cells were cultivated in the presence of vehicle, LPA (1 μM), or LPA plus a BrP-LPA (5 μM) and b TCLPA5 (5 μM) for the indicated times. Inhibitors were added 2 h prior LPA addition. Cells were stained with PE-conjugated anti-CD40, APC-conjugated anti-CD86, or PE-conjugated anti-CD206 antibodies and analyzed using a Guava easyCyte 8 Millipore flow cytometer. Results from four individual experiments in triplicate are shown as mean values + SD. (* p < 0.05; ** p < 0.01; *** p < 0.001 compared to untreated or DMSO-treated cells; # p < 0.05; ## p < 0.01; ### p < 0.001 inhibitor compared to LPA-treated cells; one-way ANOVA with Bonferroni correction)

    Techniques Used: Marker, Expressing, Staining, Flow Cytometry

    brp lpa  (Echelon Biosciences)


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    Echelon Biosciences brp lpa
    Brp Lpa, 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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    bromomethylene phosphonate lpa brp lpa  (Echelon Biosciences)


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    Echelon Biosciences bromomethylene phosphonate lpa brp lpa
    Hyperoxia increases lysophosphatidic acid <t>(LPA)</t> levels in pulmonary NKT cells in vitro. <t>BrP-LPA</t> diminishes level of LPA. Pulmonary iNKT cells were cultured and exposed to 95 % O2/5 % CO2 for 72 h with and without addition of the pan-LPA antagonist BrP-LPA. LPA levels in hyperoxia-exposed cells were significantly lower when cells were incubated with the pan LPA antagonist BrP-LPA during hyperoxia (p < 0.05). r.u. relative units (relative to the background-reaction in the absence of weight)
    Bromomethylene Phosphonate Lpa Brp Lpa, 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 "Lysophosphatidic acid generation by pulmonary NKT cell ENPP-2/autotaxin exacerbates hyperoxic lung injury"

    Article Title: Lysophosphatidic acid generation by pulmonary NKT cell ENPP-2/autotaxin exacerbates hyperoxic lung injury

    Journal: Purinergic Signalling

    doi: 10.1007/s11302-015-9463-6

    Hyperoxia increases lysophosphatidic acid (LPA) levels in pulmonary NKT cells in vitro. BrP-LPA diminishes level of LPA. Pulmonary iNKT cells were cultured and exposed to 95 % O2/5 % CO2 for 72 h with and without addition of the pan-LPA antagonist BrP-LPA. LPA levels in hyperoxia-exposed cells were significantly lower when cells were incubated with the pan LPA antagonist BrP-LPA during hyperoxia (p < 0.05). r.u. relative units (relative to the background-reaction in the absence of weight)
    Figure Legend Snippet: Hyperoxia increases lysophosphatidic acid (LPA) levels in pulmonary NKT cells in vitro. BrP-LPA diminishes level of LPA. Pulmonary iNKT cells were cultured and exposed to 95 % O2/5 % CO2 for 72 h with and without addition of the pan-LPA antagonist BrP-LPA. LPA levels in hyperoxia-exposed cells were significantly lower when cells were incubated with the pan LPA antagonist BrP-LPA during hyperoxia (p < 0.05). r.u. relative units (relative to the background-reaction in the absence of weight)

    Techniques Used: In Vitro, Cell Culture, Incubation

    Kaplan–Meier survival curves for (n > 25) and plus BrP-LPA animals after 72 h in 100 % oxygen demonstrate a clear survival benefit of those animals that received a BrP-LPA injection prior to oxygen exposure
    Figure Legend Snippet: Kaplan–Meier survival curves for (n > 25) and plus BrP-LPA animals after 72 h in 100 % oxygen demonstrate a clear survival benefit of those animals that received a BrP-LPA injection prior to oxygen exposure

    Techniques Used: Injection

    Histological assessment of hyperoxic lung injury. a H&E staining of control lung tissue and b plus BrP-LPA injection prior to hyperoxia. Animals were exposed to 100 % oxygen for 72 h. The hyperoxia-exposed lungs show severe injury with pulmonary edema and hemorrhage, whereas the BrP-LPA injected animals express milder injury
    Figure Legend Snippet: Histological assessment of hyperoxic lung injury. a H&E staining of control lung tissue and b plus BrP-LPA injection prior to hyperoxia. Animals were exposed to 100 % oxygen for 72 h. The hyperoxia-exposed lungs show severe injury with pulmonary edema and hemorrhage, whereas the BrP-LPA injected animals express milder injury

    Techniques Used: Staining, Injection

    Flow cytometry analysis of pulmonary NKT cell and PMN populations after hyperoxia. a Lungs extracted from oxygen-exposed mice with and without BrP-LPA treatment were analyzed. Baseline iNKT cell populations in the lung did not differ between BrP-LPA treated and untreated wild type mice under normoxic conditions. After 72 h of 100 % exposure, animals show significant increases of iNKT cells compared to their baseline. After BrP-LPA treatment, animals exhibit only a minimal increase of pulmonary iNKT cells in response to hyperoxia. b Populations of GR-1+/F4/80- PMN-cells increase massively after oxygen exposure in lungs. When animals were injected with BrP-LPA prior to oxygen exposure PMN-cells only increased marginally
    Figure Legend Snippet: Flow cytometry analysis of pulmonary NKT cell and PMN populations after hyperoxia. a Lungs extracted from oxygen-exposed mice with and without BrP-LPA treatment were analyzed. Baseline iNKT cell populations in the lung did not differ between BrP-LPA treated and untreated wild type mice under normoxic conditions. After 72 h of 100 % exposure, animals show significant increases of iNKT cells compared to their baseline. After BrP-LPA treatment, animals exhibit only a minimal increase of pulmonary iNKT cells in response to hyperoxia. b Populations of GR-1+/F4/80- PMN-cells increase massively after oxygen exposure in lungs. When animals were injected with BrP-LPA prior to oxygen exposure PMN-cells only increased marginally

    Techniques Used: Flow Cytometry, Injection

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    Echelon Biosciences brp lpa
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    Echelon Biosciences α bromomethylene phosphonate lpa brp lpa
    (A) HUVEC or (B) bEnd.3 cells were treated with vehicle control (▪) or 5 µM <t>BrP-LPA</t> (▴) for 45 min prior to irradiation. (C) HUVEC were treated with 0.1% fatty acid free BSA (•), 10 µM LPA (○) or 10 µM LPA plus 5 µM BrP-LPA (▪) in serum free medium for 45 min prior to irradiation. The cells were then irradiated with 0, 2, 4 and 6 Gy and plated for clonogenic survival assay. After 2–3 wks, cells were stained with 1% methylene blue and colonies consisting of >50 cells were counted by microscopy. Surviving colonies were normalized for plating efficiency. Shown are average survival fractions and SEM from three experiments; * p<0.05. (D) Equal numbers of HUVEC were plated in 96 well plates and treated with carrier control 3% fatty acid free BSA, 10 µM LPA or 10 µM LPA with 5 µM BrP-LPA in serum free medium for 45 min prior to irradiation. After 96 h, the cell viability was determined using a colorimetric cell proliferation assay (Promega). Shown is the average absorbance at 490 nm with SEM from three experiments; * p<0.05.
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    Echelon Biosciences bromo
    (A) HUVEC or (B) bEnd.3 cells were treated with vehicle control (▪) or 5 µM <t>BrP-LPA</t> (▴) for 45 min prior to irradiation. (C) HUVEC were treated with 0.1% fatty acid free BSA (•), 10 µM LPA (○) or 10 µM LPA plus 5 µM BrP-LPA (▪) in serum free medium for 45 min prior to irradiation. The cells were then irradiated with 0, 2, 4 and 6 Gy and plated for clonogenic survival assay. After 2–3 wks, cells were stained with 1% methylene blue and colonies consisting of >50 cells were counted by microscopy. Surviving colonies were normalized for plating efficiency. Shown are average survival fractions and SEM from three experiments; * p<0.05. (D) Equal numbers of HUVEC were plated in 96 well plates and treated with carrier control 3% fatty acid free BSA, 10 µM LPA or 10 µM LPA with 5 µM BrP-LPA in serum free medium for 45 min prior to irradiation. After 96 h, the cell viability was determined using a colorimetric cell proliferation assay (Promega). Shown is the average absorbance at 490 nm with SEM from three experiments; * p<0.05.
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    Echelon Biosciences bromomethylene phosphonate lpa brp lpa
    Hyperoxia increases lysophosphatidic acid <t>(LPA)</t> levels in pulmonary NKT cells in vitro. <t>BrP-LPA</t> diminishes level of LPA. Pulmonary iNKT cells were cultured and exposed to 95 % O2/5 % CO2 for 72 h with and without addition of the pan-LPA antagonist BrP-LPA. LPA levels in hyperoxia-exposed cells were significantly lower when cells were incubated with the pan LPA antagonist BrP-LPA during hyperoxia (p < 0.05). r.u. relative units (relative to the background-reaction in the absence of weight)
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    (A) HUVEC or (B) bEnd.3 cells were treated with vehicle control (▪) or 5 µM BrP-LPA (▴) for 45 min prior to irradiation. (C) HUVEC were treated with 0.1% fatty acid free BSA (•), 10 µM LPA (○) or 10 µM LPA plus 5 µM BrP-LPA (▪) in serum free medium for 45 min prior to irradiation. The cells were then irradiated with 0, 2, 4 and 6 Gy and plated for clonogenic survival assay. After 2–3 wks, cells were stained with 1% methylene blue and colonies consisting of >50 cells were counted by microscopy. Surviving colonies were normalized for plating efficiency. Shown are average survival fractions and SEM from three experiments; * p<0.05. (D) Equal numbers of HUVEC were plated in 96 well plates and treated with carrier control 3% fatty acid free BSA, 10 µM LPA or 10 µM LPA with 5 µM BrP-LPA in serum free medium for 45 min prior to irradiation. After 96 h, the cell viability was determined using a colorimetric cell proliferation assay (Promega). Shown is the average absorbance at 490 nm with SEM from three experiments; * p<0.05.

    Journal: PLoS ONE

    Article Title: Autotaxin and LPA Receptors Represent Potential Molecular Targets for the Radiosensitization of Murine Glioma through Effects on Tumor Vasculature

    doi: 10.1371/journal.pone.0022182

    Figure Lengend Snippet: (A) HUVEC or (B) bEnd.3 cells were treated with vehicle control (▪) or 5 µM BrP-LPA (▴) for 45 min prior to irradiation. (C) HUVEC were treated with 0.1% fatty acid free BSA (•), 10 µM LPA (○) or 10 µM LPA plus 5 µM BrP-LPA (▪) in serum free medium for 45 min prior to irradiation. The cells were then irradiated with 0, 2, 4 and 6 Gy and plated for clonogenic survival assay. After 2–3 wks, cells were stained with 1% methylene blue and colonies consisting of >50 cells were counted by microscopy. Surviving colonies were normalized for plating efficiency. Shown are average survival fractions and SEM from three experiments; * p<0.05. (D) Equal numbers of HUVEC were plated in 96 well plates and treated with carrier control 3% fatty acid free BSA, 10 µM LPA or 10 µM LPA with 5 µM BrP-LPA in serum free medium for 45 min prior to irradiation. After 96 h, the cell viability was determined using a colorimetric cell proliferation assay (Promega). Shown is the average absorbance at 490 nm with SEM from three experiments; * p<0.05.

    Article Snippet: To inhibit ATX and LPA receptors, we used α-bromomethylene phosphonate LPA (BrP-LPA) which was purchased from Echelon (Salt Lake City, UT).

    Techniques: Irradiation, Clonogenic Cell Survival Assay, Staining, Microscopy, Proliferation Assay

    (A, B) HUVEC or (C) bEnd.3 cells were plated on matrigel-coated 96 well plates and treated with vehicle control or 5 µM BrP-LPA for 45 min prior to irradiation with 3 Gy. (A) Shown are the representative photomicrographs of tubule formation taken 16 h after irradiation. (B) Tubule formation was quantified as number of tubules per high power field (HPF). Shown are bar graphs of mean number of tubules per HPF relative to control and SEM from three experiments; * p<0.05.

    Journal: PLoS ONE

    Article Title: Autotaxin and LPA Receptors Represent Potential Molecular Targets for the Radiosensitization of Murine Glioma through Effects on Tumor Vasculature

    doi: 10.1371/journal.pone.0022182

    Figure Lengend Snippet: (A, B) HUVEC or (C) bEnd.3 cells were plated on matrigel-coated 96 well plates and treated with vehicle control or 5 µM BrP-LPA for 45 min prior to irradiation with 3 Gy. (A) Shown are the representative photomicrographs of tubule formation taken 16 h after irradiation. (B) Tubule formation was quantified as number of tubules per high power field (HPF). Shown are bar graphs of mean number of tubules per HPF relative to control and SEM from three experiments; * p<0.05.

    Article Snippet: To inhibit ATX and LPA receptors, we used α-bromomethylene phosphonate LPA (BrP-LPA) which was purchased from Echelon (Salt Lake City, UT).

    Techniques: Irradiation

    (A) bEnd.3 or (B) HUVEC were plated on 60 mm plates and allowed to grow to 70% confluency. The semi-confluent cell layer was scraped using a sterile pipette tip to create a scratch devoid of cells. The remaining cells were treated with vehicle control or 5 µM BrP-LPA for 45 min prior to irradiation with 3 Gy. Migration was observed at 36 h. Cells were fixed with ethanol and stained with 1% methylene blue. Migrated cells were counted and normalized to surrounding cell density per HPF. Shown are representative photomicrographs and bar graphs representing the mean percentages of migrating cells relative to corresponding controls with SEM from three experiments, * p<0.05.

    Journal: PLoS ONE

    Article Title: Autotaxin and LPA Receptors Represent Potential Molecular Targets for the Radiosensitization of Murine Glioma through Effects on Tumor Vasculature

    doi: 10.1371/journal.pone.0022182

    Figure Lengend Snippet: (A) bEnd.3 or (B) HUVEC were plated on 60 mm plates and allowed to grow to 70% confluency. The semi-confluent cell layer was scraped using a sterile pipette tip to create a scratch devoid of cells. The remaining cells were treated with vehicle control or 5 µM BrP-LPA for 45 min prior to irradiation with 3 Gy. Migration was observed at 36 h. Cells were fixed with ethanol and stained with 1% methylene blue. Migrated cells were counted and normalized to surrounding cell density per HPF. Shown are representative photomicrographs and bar graphs representing the mean percentages of migrating cells relative to corresponding controls with SEM from three experiments, * p<0.05.

    Article Snippet: To inhibit ATX and LPA receptors, we used α-bromomethylene phosphonate LPA (BrP-LPA) which was purchased from Echelon (Salt Lake City, UT).

    Techniques: Transferring, Irradiation, Migration, Staining

    (A, B) Mouse glioma GL261 cells were plated on 60 mm plates and allowed to grow to 70% confluency. Plates were scraped with a pipette tip to create a scratch devoid of cells and treated with vehicle control or 5 µM BrP-LPA for 45 min before irradiation with 3 Gy. After 24 h, cells were fixed and stained with methylene blue. Migrated cells were counted and normalized to surrounding cell density per HPF. Shown are representative photomicrographs (A) and a bar graph (B) representing the mean percentages of migrating cells relative to corresponding controls with SEM from three experiments; * p<0.05. (C) For clonogenic survival assay, GL261 cells were plated and allowed to attach. After 6 h, cells were treated vehicle control or with 5 µM BrP-LPA for 45 min and irradiated with 0, 2, 4, and 6 Gy. After 10 days, surviving colonies (>50 cells) were counted and normalized for plating efficiency. Shown is the clonogenic survival curve and mean surviving fractions and SEM from three experiments; * p<0.05.

    Journal: PLoS ONE

    Article Title: Autotaxin and LPA Receptors Represent Potential Molecular Targets for the Radiosensitization of Murine Glioma through Effects on Tumor Vasculature

    doi: 10.1371/journal.pone.0022182

    Figure Lengend Snippet: (A, B) Mouse glioma GL261 cells were plated on 60 mm plates and allowed to grow to 70% confluency. Plates were scraped with a pipette tip to create a scratch devoid of cells and treated with vehicle control or 5 µM BrP-LPA for 45 min before irradiation with 3 Gy. After 24 h, cells were fixed and stained with methylene blue. Migrated cells were counted and normalized to surrounding cell density per HPF. Shown are representative photomicrographs (A) and a bar graph (B) representing the mean percentages of migrating cells relative to corresponding controls with SEM from three experiments; * p<0.05. (C) For clonogenic survival assay, GL261 cells were plated and allowed to attach. After 6 h, cells were treated vehicle control or with 5 µM BrP-LPA for 45 min and irradiated with 0, 2, 4, and 6 Gy. After 10 days, surviving colonies (>50 cells) were counted and normalized for plating efficiency. Shown is the clonogenic survival curve and mean surviving fractions and SEM from three experiments; * p<0.05.

    Article Snippet: To inhibit ATX and LPA receptors, we used α-bromomethylene phosphonate LPA (BrP-LPA) which was purchased from Echelon (Salt Lake City, UT).

    Techniques: Transferring, Irradiation, Staining, Clonogenic Cell Survival Assay

    bEnd.3 and GL261 cells were grown in co-culture for 24 h. Cells were treated with vehicle or 5 µM BrP-LPA control for 45 min before treatment with 3 Gy. Cells were lysed at 5 min after irradiation. Shown are immunoblot analyses using specific antibodies to phospho-Akt Thr308/Ser473 , total Akt, and actin. Numbers represent the ratios of phospho-Akt to total Akt protein normalized to actin (fold change relative to control).

    Journal: PLoS ONE

    Article Title: Autotaxin and LPA Receptors Represent Potential Molecular Targets for the Radiosensitization of Murine Glioma through Effects on Tumor Vasculature

    doi: 10.1371/journal.pone.0022182

    Figure Lengend Snippet: bEnd.3 and GL261 cells were grown in co-culture for 24 h. Cells were treated with vehicle or 5 µM BrP-LPA control for 45 min before treatment with 3 Gy. Cells were lysed at 5 min after irradiation. Shown are immunoblot analyses using specific antibodies to phospho-Akt Thr308/Ser473 , total Akt, and actin. Numbers represent the ratios of phospho-Akt to total Akt protein normalized to actin (fold change relative to control).

    Article Snippet: To inhibit ATX and LPA receptors, we used α-bromomethylene phosphonate LPA (BrP-LPA) which was purchased from Echelon (Salt Lake City, UT).

    Techniques: Co-Culture Assay, Irradiation, Western Blot

    GL261 cells were injected into the hind limbs of nude mice. Tumors were irradiated with 3 Gy for 5 consecutive days for a total of 15 Gy. Mice were treated with 3 mg/kg BrP-LPA or vehicle control for 45 min prior to irradiation on days 1, 3, and 5. (A) Shown are mean tumor volumes with SEM from each treatment group of 5 mice. (B) Tumor growth delay was calculated as the number of days for tumors to reach a 6-fold volume increase compared to control. Shown is a bar graph representing the mean tumor growth delay with SEM from each treatment group of 5 mice; * p<0.05.

    Journal: PLoS ONE

    Article Title: Autotaxin and LPA Receptors Represent Potential Molecular Targets for the Radiosensitization of Murine Glioma through Effects on Tumor Vasculature

    doi: 10.1371/journal.pone.0022182

    Figure Lengend Snippet: GL261 cells were injected into the hind limbs of nude mice. Tumors were irradiated with 3 Gy for 5 consecutive days for a total of 15 Gy. Mice were treated with 3 mg/kg BrP-LPA or vehicle control for 45 min prior to irradiation on days 1, 3, and 5. (A) Shown are mean tumor volumes with SEM from each treatment group of 5 mice. (B) Tumor growth delay was calculated as the number of days for tumors to reach a 6-fold volume increase compared to control. Shown is a bar graph representing the mean tumor growth delay with SEM from each treatment group of 5 mice; * p<0.05.

    Article Snippet: To inhibit ATX and LPA receptors, we used α-bromomethylene phosphonate LPA (BrP-LPA) which was purchased from Echelon (Salt Lake City, UT).

    Techniques: Injection, Irradiation

    Hyperoxia increases lysophosphatidic acid (LPA) levels in pulmonary NKT cells in vitro. BrP-LPA diminishes level of LPA. Pulmonary iNKT cells were cultured and exposed to 95 % O2/5 % CO2 for 72 h with and without addition of the pan-LPA antagonist BrP-LPA. LPA levels in hyperoxia-exposed cells were significantly lower when cells were incubated with the pan LPA antagonist BrP-LPA during hyperoxia (p < 0.05). r.u. relative units (relative to the background-reaction in the absence of weight)

    Journal: Purinergic Signalling

    Article Title: Lysophosphatidic acid generation by pulmonary NKT cell ENPP-2/autotaxin exacerbates hyperoxic lung injury

    doi: 10.1007/s11302-015-9463-6

    Figure Lengend Snippet: Hyperoxia increases lysophosphatidic acid (LPA) levels in pulmonary NKT cells in vitro. BrP-LPA diminishes level of LPA. Pulmonary iNKT cells were cultured and exposed to 95 % O2/5 % CO2 for 72 h with and without addition of the pan-LPA antagonist BrP-LPA. LPA levels in hyperoxia-exposed cells were significantly lower when cells were incubated with the pan LPA antagonist BrP-LPA during hyperoxia (p < 0.05). r.u. relative units (relative to the background-reaction in the absence of weight)

    Article Snippet: Bromomethylene phosphonate LPA (BrP-LPA) was purchased from Echelon®.

    Techniques: In Vitro, Cell Culture, Incubation

    Kaplan–Meier survival curves for (n > 25) and plus BrP-LPA animals after 72 h in 100 % oxygen demonstrate a clear survival benefit of those animals that received a BrP-LPA injection prior to oxygen exposure

    Journal: Purinergic Signalling

    Article Title: Lysophosphatidic acid generation by pulmonary NKT cell ENPP-2/autotaxin exacerbates hyperoxic lung injury

    doi: 10.1007/s11302-015-9463-6

    Figure Lengend Snippet: Kaplan–Meier survival curves for (n > 25) and plus BrP-LPA animals after 72 h in 100 % oxygen demonstrate a clear survival benefit of those animals that received a BrP-LPA injection prior to oxygen exposure

    Article Snippet: Bromomethylene phosphonate LPA (BrP-LPA) was purchased from Echelon®.

    Techniques: Injection

    Histological assessment of hyperoxic lung injury. a H&E staining of control lung tissue and b plus BrP-LPA injection prior to hyperoxia. Animals were exposed to 100 % oxygen for 72 h. The hyperoxia-exposed lungs show severe injury with pulmonary edema and hemorrhage, whereas the BrP-LPA injected animals express milder injury

    Journal: Purinergic Signalling

    Article Title: Lysophosphatidic acid generation by pulmonary NKT cell ENPP-2/autotaxin exacerbates hyperoxic lung injury

    doi: 10.1007/s11302-015-9463-6

    Figure Lengend Snippet: Histological assessment of hyperoxic lung injury. a H&E staining of control lung tissue and b plus BrP-LPA injection prior to hyperoxia. Animals were exposed to 100 % oxygen for 72 h. The hyperoxia-exposed lungs show severe injury with pulmonary edema and hemorrhage, whereas the BrP-LPA injected animals express milder injury

    Article Snippet: Bromomethylene phosphonate LPA (BrP-LPA) was purchased from Echelon®.

    Techniques: Staining, Injection

    Flow cytometry analysis of pulmonary NKT cell and PMN populations after hyperoxia. a Lungs extracted from oxygen-exposed mice with and without BrP-LPA treatment were analyzed. Baseline iNKT cell populations in the lung did not differ between BrP-LPA treated and untreated wild type mice under normoxic conditions. After 72 h of 100 % exposure, animals show significant increases of iNKT cells compared to their baseline. After BrP-LPA treatment, animals exhibit only a minimal increase of pulmonary iNKT cells in response to hyperoxia. b Populations of GR-1+/F4/80- PMN-cells increase massively after oxygen exposure in lungs. When animals were injected with BrP-LPA prior to oxygen exposure PMN-cells only increased marginally

    Journal: Purinergic Signalling

    Article Title: Lysophosphatidic acid generation by pulmonary NKT cell ENPP-2/autotaxin exacerbates hyperoxic lung injury

    doi: 10.1007/s11302-015-9463-6

    Figure Lengend Snippet: Flow cytometry analysis of pulmonary NKT cell and PMN populations after hyperoxia. a Lungs extracted from oxygen-exposed mice with and without BrP-LPA treatment were analyzed. Baseline iNKT cell populations in the lung did not differ between BrP-LPA treated and untreated wild type mice under normoxic conditions. After 72 h of 100 % exposure, animals show significant increases of iNKT cells compared to their baseline. After BrP-LPA treatment, animals exhibit only a minimal increase of pulmonary iNKT cells in response to hyperoxia. b Populations of GR-1+/F4/80- PMN-cells increase massively after oxygen exposure in lungs. When animals were injected with BrP-LPA prior to oxygen exposure PMN-cells only increased marginally

    Article Snippet: Bromomethylene phosphonate LPA (BrP-LPA) was purchased from Echelon®.

    Techniques: Flow Cytometry, Injection