sphingosine 1 phosphate elisa kit  (Echelon Biosciences)


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    Echelon Biosciences sphingosine 1 phosphate elisa kit
    Sphingosine 1 Phosphate Elisa Kit, 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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    sphingosine 1 phoshate s1p  (Echelon Biosciences)


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    Echelon Biosciences sphingosine 1 phoshate s1p
    <t> Sphingosine-1-phosphate </t> levels in HDL, HDL2, and HDL3.
    Sphingosine 1 Phoshate S1p, 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 "eNOS Activation by HDL Is Impaired in Genetic CETP Deficiency"

    Article Title: eNOS Activation by HDL Is Impaired in Genetic CETP Deficiency

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0095925

     Sphingosine-1-phosphate  levels in HDL, HDL2, and HDL3.
    Figure Legend Snippet: Sphingosine-1-phosphate levels in HDL, HDL2, and HDL3.

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    s1p  (Echelon Biosciences)


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    Echelon Biosciences s1p
    Oral keratinocytes were incubated with or without Sphk-1 inhibitor (2 µM) for 2 h prior to challenging the cells with various TLR agonists namely, heat inactivated P. gingivalis (MOI:100), FSL-1 (1 µg/ml), Pam3CSK4 (0.5 µg/ml), P.gingivalis LPS (1 µg/ml), E. Coli LPS (1 µg/ml), ssRNA (0.1 µg/ml), Poly I:C (5 µg/ml) ODN (0.5 µg/ml), Imiquimod (0.1 µg/ml), Flagellin (0.25 µg/mL) ( A ); the IL-1α (2.5 ng/ml), IL-1β (2.5 ng/ml) and TNF-α (2.5 ng/ml) ( B ) and GPCR agonist <t>S1P</t> (100 nM) ( C ) for 24 h. The supernatant was collected after 24 h and HBD-2 ELISA was performed using Human BD-2 ELISA kit. The Sphk-1 inhibitor ablated HBD-2 induction with the agonists tested. The time course experiment was performed by pretreating the cells with Sphk-1 (2 µg/ml) for 2 h before challenging with FSL-1 (1 µg/ml) for 0, 30, 60, 90, 120 and 240 min. The total protein was collected and subjected to immunoblot with ser225 phospho specific Sphk-1 antibody and β-actin as loading control. We noted increase in the phosphorylation level of Sphk-1 at ser225 as early as 60 min and the level of phosphorylation was down regulated in the presence of Sphk-1 inhibitor ( D ). Control cells received DMSO unless otherwise stated. Results are mean ± SEM and are representative of three independent experiments. Statistical comparisons are shown by horizontal bars with asterisks above them (* indicates p<0.05 determined by ANOVA and Tukey multiple comparison test).
    S1p, 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 "Sphingosine Kinase-1 Is Required for Toll Mediated β-Defensin 2 Induction in Human Oral Keratinocytes"

    Article Title: Sphingosine Kinase-1 Is Required for Toll Mediated β-Defensin 2 Induction in Human Oral Keratinocytes

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0011512

    Oral keratinocytes were incubated with or without Sphk-1 inhibitor (2 µM) for 2 h prior to challenging the cells with various TLR agonists namely, heat inactivated P. gingivalis (MOI:100), FSL-1 (1 µg/ml), Pam3CSK4 (0.5 µg/ml), P.gingivalis LPS (1 µg/ml), E. Coli LPS (1 µg/ml), ssRNA (0.1 µg/ml), Poly I:C (5 µg/ml) ODN (0.5 µg/ml), Imiquimod (0.1 µg/ml), Flagellin (0.25 µg/mL) ( A ); the IL-1α (2.5 ng/ml), IL-1β (2.5 ng/ml) and TNF-α (2.5 ng/ml) ( B ) and GPCR agonist S1P (100 nM) ( C ) for 24 h. The supernatant was collected after 24 h and HBD-2 ELISA was performed using Human BD-2 ELISA kit. The Sphk-1 inhibitor ablated HBD-2 induction with the agonists tested. The time course experiment was performed by pretreating the cells with Sphk-1 (2 µg/ml) for 2 h before challenging with FSL-1 (1 µg/ml) for 0, 30, 60, 90, 120 and 240 min. The total protein was collected and subjected to immunoblot with ser225 phospho specific Sphk-1 antibody and β-actin as loading control. We noted increase in the phosphorylation level of Sphk-1 at ser225 as early as 60 min and the level of phosphorylation was down regulated in the presence of Sphk-1 inhibitor ( D ). Control cells received DMSO unless otherwise stated. Results are mean ± SEM and are representative of three independent experiments. Statistical comparisons are shown by horizontal bars with asterisks above them (* indicates p<0.05 determined by ANOVA and Tukey multiple comparison test).
    Figure Legend Snippet: Oral keratinocytes were incubated with or without Sphk-1 inhibitor (2 µM) for 2 h prior to challenging the cells with various TLR agonists namely, heat inactivated P. gingivalis (MOI:100), FSL-1 (1 µg/ml), Pam3CSK4 (0.5 µg/ml), P.gingivalis LPS (1 µg/ml), E. Coli LPS (1 µg/ml), ssRNA (0.1 µg/ml), Poly I:C (5 µg/ml) ODN (0.5 µg/ml), Imiquimod (0.1 µg/ml), Flagellin (0.25 µg/mL) ( A ); the IL-1α (2.5 ng/ml), IL-1β (2.5 ng/ml) and TNF-α (2.5 ng/ml) ( B ) and GPCR agonist S1P (100 nM) ( C ) for 24 h. The supernatant was collected after 24 h and HBD-2 ELISA was performed using Human BD-2 ELISA kit. The Sphk-1 inhibitor ablated HBD-2 induction with the agonists tested. The time course experiment was performed by pretreating the cells with Sphk-1 (2 µg/ml) for 2 h before challenging with FSL-1 (1 µg/ml) for 0, 30, 60, 90, 120 and 240 min. The total protein was collected and subjected to immunoblot with ser225 phospho specific Sphk-1 antibody and β-actin as loading control. We noted increase in the phosphorylation level of Sphk-1 at ser225 as early as 60 min and the level of phosphorylation was down regulated in the presence of Sphk-1 inhibitor ( D ). Control cells received DMSO unless otherwise stated. Results are mean ± SEM and are representative of three independent experiments. Statistical comparisons are shown by horizontal bars with asterisks above them (* indicates p<0.05 determined by ANOVA and Tukey multiple comparison test).

    Techniques Used: Incubation, Enzyme-linked Immunosorbent Assay, Western Blot

    The cells were transiently transfected with siRNA against Sphk-1 and incubated with FSL-1 (1 µg/ml), Imiquimod (0.1 µg/ml) and IL-1β (2.5 ng/ml). The supernatant was collected after 24 h challenge and subjected to human HBD-2 ELISA. siRNA against Sphk-1 down modulated agonist induced HBD-2 in HGECs ( A ). Intracellular S1P was measured using S1P ELISA kit after challenging with FSL-1 (1 µg/ml) for 2 h in cells pre-incubated with Sphk-1 inhibitor. The reaction was terminated after 2 h and 100 µg of total protein was subjected to ELISA. The intracellular S1P production was downregulated by Sphk-1 inhibitor ( B ). Total RNA was collected and converted to cDNA from the cells challenged with FSL-1 (1 µg/ml) for 24 h. cDNA was subjected to real time PCR with Sphk-1, Sphk-2 and GAPDH endogenous control TaqMan probes. Sphk-1 mRNA expression was highly upregulated compared to Sphk2 mRNA expression showing Sphk-1 as a predominant kinase in HGECs upon ligand challenge ( C ). Control cells received DMSO unless otherwise stated. Results are mean ± SEM and are representative of three independent experiments. Statistical comparisons are shown by horizontal bars with asterisks above them (* indicates p<0.05 determined by ANOVA and Tukey multiple comparison test).
    Figure Legend Snippet: The cells were transiently transfected with siRNA against Sphk-1 and incubated with FSL-1 (1 µg/ml), Imiquimod (0.1 µg/ml) and IL-1β (2.5 ng/ml). The supernatant was collected after 24 h challenge and subjected to human HBD-2 ELISA. siRNA against Sphk-1 down modulated agonist induced HBD-2 in HGECs ( A ). Intracellular S1P was measured using S1P ELISA kit after challenging with FSL-1 (1 µg/ml) for 2 h in cells pre-incubated with Sphk-1 inhibitor. The reaction was terminated after 2 h and 100 µg of total protein was subjected to ELISA. The intracellular S1P production was downregulated by Sphk-1 inhibitor ( B ). Total RNA was collected and converted to cDNA from the cells challenged with FSL-1 (1 µg/ml) for 24 h. cDNA was subjected to real time PCR with Sphk-1, Sphk-2 and GAPDH endogenous control TaqMan probes. Sphk-1 mRNA expression was highly upregulated compared to Sphk2 mRNA expression showing Sphk-1 as a predominant kinase in HGECs upon ligand challenge ( C ). Control cells received DMSO unless otherwise stated. Results are mean ± SEM and are representative of three independent experiments. Statistical comparisons are shown by horizontal bars with asterisks above them (* indicates p<0.05 determined by ANOVA and Tukey multiple comparison test).

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

    s1p elisa kit  (Echelon Biosciences)


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    Echelon Biosciences s1p elisa kit
    S1p Elisa Kit, 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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    sphingosine 1 phosphase elisa kit  (Echelon Biosciences)


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    Echelon Biosciences sphingosine 1 phosphase elisa kit
    Correlative plasma markers (A) acid sphingomyelinase (ASM); and (B) <t>sphingosine-1-phosphate</t> <t>(S1P).</t>
    Sphingosine 1 Phosphase Elisa Kit, 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 "Quantifying the Changes in the Tumour Vascular Micro-environment in Spinal Metastases Treated with Stereotactic Body Radiotherapy - A Single Arm Prospective Study"

    Article Title: Quantifying the Changes in the Tumour Vascular Micro-environment in Spinal Metastases Treated with Stereotactic Body Radiotherapy - A Single Arm Prospective Study

    Journal: Radiology and Oncology

    doi: 10.2478/raon-2022-0046

    Correlative plasma markers (A) acid sphingomyelinase (ASM); and (B) sphingosine-1-phosphate (S1P).
    Figure Legend Snippet: Correlative plasma markers (A) acid sphingomyelinase (ASM); and (B) sphingosine-1-phosphate (S1P).

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    s1p elisa kit  (Echelon Biosciences)


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    Echelon Biosciences s1p elisa kit
    S1p Elisa Kit, 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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    s1p elisa kits  (Echelon Biosciences)


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    Echelon Biosciences s1p elisa kits
    S1p Elisa Kits, 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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    immunosorbent assay elisa  (Echelon Biosciences)


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    Echelon Biosciences immunosorbent assay elisa
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    sphingosine 1 phosphate assay kit  (Echelon Biosciences)


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    Echelon Biosciences sphingosine 1 phosphate assay kit
    Sphingosine 1 Phosphate Assay Kit, 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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    s1p assay kit  (Echelon Biosciences)


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    Echelon Biosciences s1p assay kit
    Effects of Sal on the expressions of <t>SphK/S1P/S1PRs</t> signaling pathway in mice liver induced by CCl 4 . (A) : The mRNA expressions of SphK1, SphK2, SPPR1, S1PR2 and S1PR3 in mice liver tissue. (B) : Level of S1P in serum from mice. (C) : The protein expressions of SphK1, SphK2, of S1PR1, S1PR2, and S1PR3 in mice liver tissue ( n = 15, * p < 0.05, ** p < 0.01, vs Cr; # p < 0.05, ## p < 0.01, vs. M.) SphK1: Sphingosine kinase 1; S1P: Sphingosine-1-phosphate; S1PR1: Sphingosine 1-phosphate receptor 1; S1PR2: Sphingosine 1-phosphate receptor 2; S1PR3: Sphingosine 1-phosphate receptor 3.
    S1p Assay Kit, 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 "Salidroside Inhibits CCl 4 -Induced Liver Fibrosis in Mice by Reducing Activation and Migration of HSC Induced by Liver Sinusoidal Endothelial Cell-Derived Exosomal SphK1"

    Article Title: Salidroside Inhibits CCl 4 -Induced Liver Fibrosis in Mice by Reducing Activation and Migration of HSC Induced by Liver Sinusoidal Endothelial Cell-Derived Exosomal SphK1

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2021.677810

    Effects of Sal on the expressions of SphK/S1P/S1PRs signaling pathway in mice liver induced by CCl 4 . (A) : The mRNA expressions of SphK1, SphK2, SPPR1, S1PR2 and S1PR3 in mice liver tissue. (B) : Level of S1P in serum from mice. (C) : The protein expressions of SphK1, SphK2, of S1PR1, S1PR2, and S1PR3 in mice liver tissue ( n = 15, * p < 0.05, ** p < 0.01, vs Cr; # p < 0.05, ## p < 0.01, vs. M.) SphK1: Sphingosine kinase 1; S1P: Sphingosine-1-phosphate; S1PR1: Sphingosine 1-phosphate receptor 1; S1PR2: Sphingosine 1-phosphate receptor 2; S1PR3: Sphingosine 1-phosphate receptor 3.
    Figure Legend Snippet: Effects of Sal on the expressions of SphK/S1P/S1PRs signaling pathway in mice liver induced by CCl 4 . (A) : The mRNA expressions of SphK1, SphK2, SPPR1, S1PR2 and S1PR3 in mice liver tissue. (B) : Level of S1P in serum from mice. (C) : The protein expressions of SphK1, SphK2, of S1PR1, S1PR2, and S1PR3 in mice liver tissue ( n = 15, * p < 0.05, ** p < 0.01, vs Cr; # p < 0.05, ## p < 0.01, vs. M.) SphK1: Sphingosine kinase 1; S1P: Sphingosine-1-phosphate; S1PR1: Sphingosine 1-phosphate receptor 1; S1PR2: Sphingosine 1-phosphate receptor 2; S1PR3: Sphingosine 1-phosphate receptor 3.

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    Echelon Biosciences sphingosine 1 phosphate elisa kit
    Sphingosine 1 Phosphate Elisa Kit, 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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    Oral keratinocytes were incubated with or without Sphk-1 inhibitor (2 µM) for 2 h prior to challenging the cells with various TLR agonists namely, heat inactivated P. gingivalis (MOI:100), FSL-1 (1 µg/ml), Pam3CSK4 (0.5 µg/ml), P.gingivalis LPS (1 µg/ml), E. Coli LPS (1 µg/ml), ssRNA (0.1 µg/ml), Poly I:C (5 µg/ml) ODN (0.5 µg/ml), Imiquimod (0.1 µg/ml), Flagellin (0.25 µg/mL) ( A ); the IL-1α (2.5 ng/ml), IL-1β (2.5 ng/ml) and TNF-α (2.5 ng/ml) ( B ) and GPCR agonist <t>S1P</t> (100 nM) ( C ) for 24 h. The supernatant was collected after 24 h and HBD-2 ELISA was performed using Human BD-2 ELISA kit. The Sphk-1 inhibitor ablated HBD-2 induction with the agonists tested. The time course experiment was performed by pretreating the cells with Sphk-1 (2 µg/ml) for 2 h before challenging with FSL-1 (1 µg/ml) for 0, 30, 60, 90, 120 and 240 min. The total protein was collected and subjected to immunoblot with ser225 phospho specific Sphk-1 antibody and β-actin as loading control. We noted increase in the phosphorylation level of Sphk-1 at ser225 as early as 60 min and the level of phosphorylation was down regulated in the presence of Sphk-1 inhibitor ( D ). Control cells received DMSO unless otherwise stated. Results are mean ± SEM and are representative of three independent experiments. Statistical comparisons are shown by horizontal bars with asterisks above them (* indicates p<0.05 determined by ANOVA and Tukey multiple comparison test).
    S1p, 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 s1p elisa kit
    Oral keratinocytes were incubated with or without Sphk-1 inhibitor (2 µM) for 2 h prior to challenging the cells with various TLR agonists namely, heat inactivated P. gingivalis (MOI:100), FSL-1 (1 µg/ml), Pam3CSK4 (0.5 µg/ml), P.gingivalis LPS (1 µg/ml), E. Coli LPS (1 µg/ml), ssRNA (0.1 µg/ml), Poly I:C (5 µg/ml) ODN (0.5 µg/ml), Imiquimod (0.1 µg/ml), Flagellin (0.25 µg/mL) ( A ); the IL-1α (2.5 ng/ml), IL-1β (2.5 ng/ml) and TNF-α (2.5 ng/ml) ( B ) and GPCR agonist <t>S1P</t> (100 nM) ( C ) for 24 h. The supernatant was collected after 24 h and HBD-2 ELISA was performed using Human BD-2 ELISA kit. The Sphk-1 inhibitor ablated HBD-2 induction with the agonists tested. The time course experiment was performed by pretreating the cells with Sphk-1 (2 µg/ml) for 2 h before challenging with FSL-1 (1 µg/ml) for 0, 30, 60, 90, 120 and 240 min. The total protein was collected and subjected to immunoblot with ser225 phospho specific Sphk-1 antibody and β-actin as loading control. We noted increase in the phosphorylation level of Sphk-1 at ser225 as early as 60 min and the level of phosphorylation was down regulated in the presence of Sphk-1 inhibitor ( D ). Control cells received DMSO unless otherwise stated. Results are mean ± SEM and are representative of three independent experiments. Statistical comparisons are shown by horizontal bars with asterisks above them (* indicates p<0.05 determined by ANOVA and Tukey multiple comparison test).
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    Correlative plasma markers (A) acid sphingomyelinase (ASM); and (B) <t>sphingosine-1-phosphate</t> <t>(S1P).</t>
    Sphingosine 1 Phosphase Elisa Kit, 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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    Correlative plasma markers (A) acid sphingomyelinase (ASM); and (B) <t>sphingosine-1-phosphate</t> <t>(S1P).</t>
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    93
    Echelon Biosciences sphingosine 1 phosphate assay kit
    Correlative plasma markers (A) acid sphingomyelinase (ASM); and (B) <t>sphingosine-1-phosphate</t> <t>(S1P).</t>
    Sphingosine 1 Phosphate Assay Kit, 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/sphingosine 1 phosphate assay kit/product/Echelon Biosciences
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sphingosine 1 phosphate assay kit - by Bioz Stars, 2024-05
    93/100 stars
      Buy from Supplier

    93
    Echelon Biosciences s1p assay kit
    Effects of Sal on the expressions of <t>SphK/S1P/S1PRs</t> signaling pathway in mice liver induced by CCl 4 . (A) : The mRNA expressions of SphK1, SphK2, SPPR1, S1PR2 and S1PR3 in mice liver tissue. (B) : Level of S1P in serum from mice. (C) : The protein expressions of SphK1, SphK2, of S1PR1, S1PR2, and S1PR3 in mice liver tissue ( n = 15, * p < 0.05, ** p < 0.01, vs Cr; # p < 0.05, ## p < 0.01, vs. M.) SphK1: Sphingosine kinase 1; S1P: Sphingosine-1-phosphate; S1PR1: Sphingosine 1-phosphate receptor 1; S1PR2: Sphingosine 1-phosphate receptor 2; S1PR3: Sphingosine 1-phosphate receptor 3.
    S1p Assay Kit, 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/s1p assay kit/product/Echelon Biosciences
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    s1p assay kit - by Bioz Stars, 2024-05
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    Image Search Results


     Sphingosine-1-phosphate  levels in HDL, HDL2, and HDL3.

    Journal: PLoS ONE

    Article Title: eNOS Activation by HDL Is Impaired in Genetic CETP Deficiency

    doi: 10.1371/journal.pone.0095925

    Figure Lengend Snippet: Sphingosine-1-phosphate levels in HDL, HDL2, and HDL3.

    Article Snippet: The concentration of sphingosine-1-phoshate (S1P) in isolated HDL fractions was measured with a commercial competitive ELISA kit (Echelon Biosciences Inc., Salt Lake City, UT, USA) and normalized by protein concentration.

    Techniques:

    Oral keratinocytes were incubated with or without Sphk-1 inhibitor (2 µM) for 2 h prior to challenging the cells with various TLR agonists namely, heat inactivated P. gingivalis (MOI:100), FSL-1 (1 µg/ml), Pam3CSK4 (0.5 µg/ml), P.gingivalis LPS (1 µg/ml), E. Coli LPS (1 µg/ml), ssRNA (0.1 µg/ml), Poly I:C (5 µg/ml) ODN (0.5 µg/ml), Imiquimod (0.1 µg/ml), Flagellin (0.25 µg/mL) ( A ); the IL-1α (2.5 ng/ml), IL-1β (2.5 ng/ml) and TNF-α (2.5 ng/ml) ( B ) and GPCR agonist S1P (100 nM) ( C ) for 24 h. The supernatant was collected after 24 h and HBD-2 ELISA was performed using Human BD-2 ELISA kit. The Sphk-1 inhibitor ablated HBD-2 induction with the agonists tested. The time course experiment was performed by pretreating the cells with Sphk-1 (2 µg/ml) for 2 h before challenging with FSL-1 (1 µg/ml) for 0, 30, 60, 90, 120 and 240 min. The total protein was collected and subjected to immunoblot with ser225 phospho specific Sphk-1 antibody and β-actin as loading control. We noted increase in the phosphorylation level of Sphk-1 at ser225 as early as 60 min and the level of phosphorylation was down regulated in the presence of Sphk-1 inhibitor ( D ). Control cells received DMSO unless otherwise stated. Results are mean ± SEM and are representative of three independent experiments. Statistical comparisons are shown by horizontal bars with asterisks above them (* indicates p<0.05 determined by ANOVA and Tukey multiple comparison test).

    Journal: PLoS ONE

    Article Title: Sphingosine Kinase-1 Is Required for Toll Mediated β-Defensin 2 Induction in Human Oral Keratinocytes

    doi: 10.1371/journal.pone.0011512

    Figure Lengend Snippet: Oral keratinocytes were incubated with or without Sphk-1 inhibitor (2 µM) for 2 h prior to challenging the cells with various TLR agonists namely, heat inactivated P. gingivalis (MOI:100), FSL-1 (1 µg/ml), Pam3CSK4 (0.5 µg/ml), P.gingivalis LPS (1 µg/ml), E. Coli LPS (1 µg/ml), ssRNA (0.1 µg/ml), Poly I:C (5 µg/ml) ODN (0.5 µg/ml), Imiquimod (0.1 µg/ml), Flagellin (0.25 µg/mL) ( A ); the IL-1α (2.5 ng/ml), IL-1β (2.5 ng/ml) and TNF-α (2.5 ng/ml) ( B ) and GPCR agonist S1P (100 nM) ( C ) for 24 h. The supernatant was collected after 24 h and HBD-2 ELISA was performed using Human BD-2 ELISA kit. The Sphk-1 inhibitor ablated HBD-2 induction with the agonists tested. The time course experiment was performed by pretreating the cells with Sphk-1 (2 µg/ml) for 2 h before challenging with FSL-1 (1 µg/ml) for 0, 30, 60, 90, 120 and 240 min. The total protein was collected and subjected to immunoblot with ser225 phospho specific Sphk-1 antibody and β-actin as loading control. We noted increase in the phosphorylation level of Sphk-1 at ser225 as early as 60 min and the level of phosphorylation was down regulated in the presence of Sphk-1 inhibitor ( D ). Control cells received DMSO unless otherwise stated. Results are mean ± SEM and are representative of three independent experiments. Statistical comparisons are shown by horizontal bars with asterisks above them (* indicates p<0.05 determined by ANOVA and Tukey multiple comparison test).

    Article Snippet: TransAM NF-kB p65 kit was purchased from Active Motif, CA and the ELISA kit for S1P was procured from Echelon Biosciences, UT, HBD-2 ELISA kit was from Peprotech, NJ and IL-6, IL-1β and TNF-α were from BD biosciences, CA.

    Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Western Blot

    The cells were transiently transfected with siRNA against Sphk-1 and incubated with FSL-1 (1 µg/ml), Imiquimod (0.1 µg/ml) and IL-1β (2.5 ng/ml). The supernatant was collected after 24 h challenge and subjected to human HBD-2 ELISA. siRNA against Sphk-1 down modulated agonist induced HBD-2 in HGECs ( A ). Intracellular S1P was measured using S1P ELISA kit after challenging with FSL-1 (1 µg/ml) for 2 h in cells pre-incubated with Sphk-1 inhibitor. The reaction was terminated after 2 h and 100 µg of total protein was subjected to ELISA. The intracellular S1P production was downregulated by Sphk-1 inhibitor ( B ). Total RNA was collected and converted to cDNA from the cells challenged with FSL-1 (1 µg/ml) for 24 h. cDNA was subjected to real time PCR with Sphk-1, Sphk-2 and GAPDH endogenous control TaqMan probes. Sphk-1 mRNA expression was highly upregulated compared to Sphk2 mRNA expression showing Sphk-1 as a predominant kinase in HGECs upon ligand challenge ( C ). Control cells received DMSO unless otherwise stated. Results are mean ± SEM and are representative of three independent experiments. Statistical comparisons are shown by horizontal bars with asterisks above them (* indicates p<0.05 determined by ANOVA and Tukey multiple comparison test).

    Journal: PLoS ONE

    Article Title: Sphingosine Kinase-1 Is Required for Toll Mediated β-Defensin 2 Induction in Human Oral Keratinocytes

    doi: 10.1371/journal.pone.0011512

    Figure Lengend Snippet: The cells were transiently transfected with siRNA against Sphk-1 and incubated with FSL-1 (1 µg/ml), Imiquimod (0.1 µg/ml) and IL-1β (2.5 ng/ml). The supernatant was collected after 24 h challenge and subjected to human HBD-2 ELISA. siRNA against Sphk-1 down modulated agonist induced HBD-2 in HGECs ( A ). Intracellular S1P was measured using S1P ELISA kit after challenging with FSL-1 (1 µg/ml) for 2 h in cells pre-incubated with Sphk-1 inhibitor. The reaction was terminated after 2 h and 100 µg of total protein was subjected to ELISA. The intracellular S1P production was downregulated by Sphk-1 inhibitor ( B ). Total RNA was collected and converted to cDNA from the cells challenged with FSL-1 (1 µg/ml) for 24 h. cDNA was subjected to real time PCR with Sphk-1, Sphk-2 and GAPDH endogenous control TaqMan probes. Sphk-1 mRNA expression was highly upregulated compared to Sphk2 mRNA expression showing Sphk-1 as a predominant kinase in HGECs upon ligand challenge ( C ). Control cells received DMSO unless otherwise stated. Results are mean ± SEM and are representative of three independent experiments. Statistical comparisons are shown by horizontal bars with asterisks above them (* indicates p<0.05 determined by ANOVA and Tukey multiple comparison test).

    Article Snippet: TransAM NF-kB p65 kit was purchased from Active Motif, CA and the ELISA kit for S1P was procured from Echelon Biosciences, UT, HBD-2 ELISA kit was from Peprotech, NJ and IL-6, IL-1β and TNF-α were from BD biosciences, CA.

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

    Correlative plasma markers (A) acid sphingomyelinase (ASM); and (B) sphingosine-1-phosphate (S1P).

    Journal: Radiology and Oncology

    Article Title: Quantifying the Changes in the Tumour Vascular Micro-environment in Spinal Metastases Treated with Stereotactic Body Radiotherapy - A Single Arm Prospective Study

    doi: 10.2478/raon-2022-0046

    Figure Lengend Snippet: Correlative plasma markers (A) acid sphingomyelinase (ASM); and (B) sphingosine-1-phosphate (S1P).

    Article Snippet: Samples were centrifuged for 15 minutes at 1000×g at 2 - 8 o C within 30 minutes of collection, and plasma was removed to be stored at -80 o C. Once plasma was collected from all patients, ASM (Human Acid Sphingomyelinase ELISA Kit, Elabscience) and S1P (Sphingosine 1-phosphase ELISA kit, Echelon Biosciences, UT, USA) was batch evaluated using a semi-quantitative method, using the Sandwich-ELISA method.

    Techniques:

    Effects of Sal on the expressions of SphK/S1P/S1PRs signaling pathway in mice liver induced by CCl 4 . (A) : The mRNA expressions of SphK1, SphK2, SPPR1, S1PR2 and S1PR3 in mice liver tissue. (B) : Level of S1P in serum from mice. (C) : The protein expressions of SphK1, SphK2, of S1PR1, S1PR2, and S1PR3 in mice liver tissue ( n = 15, * p < 0.05, ** p < 0.01, vs Cr; # p < 0.05, ## p < 0.01, vs. M.) SphK1: Sphingosine kinase 1; S1P: Sphingosine-1-phosphate; S1PR1: Sphingosine 1-phosphate receptor 1; S1PR2: Sphingosine 1-phosphate receptor 2; S1PR3: Sphingosine 1-phosphate receptor 3.

    Journal: Frontiers in Pharmacology

    Article Title: Salidroside Inhibits CCl 4 -Induced Liver Fibrosis in Mice by Reducing Activation and Migration of HSC Induced by Liver Sinusoidal Endothelial Cell-Derived Exosomal SphK1

    doi: 10.3389/fphar.2021.677810

    Figure Lengend Snippet: Effects of Sal on the expressions of SphK/S1P/S1PRs signaling pathway in mice liver induced by CCl 4 . (A) : The mRNA expressions of SphK1, SphK2, SPPR1, S1PR2 and S1PR3 in mice liver tissue. (B) : Level of S1P in serum from mice. (C) : The protein expressions of SphK1, SphK2, of S1PR1, S1PR2, and S1PR3 in mice liver tissue ( n = 15, * p < 0.05, ** p < 0.01, vs Cr; # p < 0.05, ## p < 0.01, vs. M.) SphK1: Sphingosine kinase 1; S1P: Sphingosine-1-phosphate; S1PR1: Sphingosine 1-phosphate receptor 1; S1PR2: Sphingosine 1-phosphate receptor 2; S1PR3: Sphingosine 1-phosphate receptor 3.

    Article Snippet: The S1P Assay Kit (S1P-ELISA) was delivered by Echelon Biosciences (Salt Lake City, United States).

    Techniques: