s1p assay kit  (Echelon Biosciences)


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    Echelon Biosciences s1p assay kit
    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) <t>S1P</t> was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.
    S1p Assay Kit, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    s1p assay kit - by Bioz Stars, 2023-01
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    Images

    1) Product Images from "Sphingosine Kinase-2 Maintains Viral Latency and Survival for KSHV-Infected Endothelial Cells"

    Article Title: Sphingosine Kinase-2 Maintains Viral Latency and Survival for KSHV-Infected Endothelial Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0102314

    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.
    Figure Legend Snippet: ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.

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

    s1p assay kit  (Echelon Biosciences)


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    Echelon Biosciences s1p assay kit
    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) <t>S1P</t> was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.
    S1p Assay Kit, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 95/100, 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 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    s1p assay kit - by Bioz Stars, 2023-01
    95/100 stars

    Images

    1) Product Images from "Sphingosine Kinase-2 Maintains Viral Latency and Survival for KSHV-Infected Endothelial Cells"

    Article Title: Sphingosine Kinase-2 Maintains Viral Latency and Survival for KSHV-Infected Endothelial Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0102314

    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.
    Figure Legend Snippet: ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.

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

    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: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 1 article reviews
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    s1p - by Bioz Stars, 2023-01
    95/100 stars

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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 assay kit  (Echelon Biosciences)


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    Echelon Biosciences s1p assay kit
    Differential involvement of <t>S1P</t> receptors in S1P- and CoCl 2 -mediated chemokine secretion by normal FLS and RAFLS. Human primary FLS from normal ( n = 4) and RA ( n = 4) donors were incubated with 5 μ M S1P (a, b) or 200 μ M CoCl 2 (c, d). Where indicated, cells were pretreated with S1P 3 antagonist CAY10444 (5 μ M) or S1P 2 antagonist JTE-013 (5 μ M) for 30 min before stimulation with S1P or CoCl 2 . The amounts of chemokines released in the supernatants were monitored after 24 h. Data are expressed as percentage of chemokine production induced by S1P (a, b) or CoCl 2 (c, d). The data are the means ± SE from four experiments (4 different donors) performed in triplicate (3 independent experiments). For statistical comparative analyses, the samples stimulated with S1P (a, b) or CoCl 2 (c, d) were compared to those stimulated in the presence of CAY10444 or JTE-013, respectively. ∗∗ p < 0.01; ∗∗∗ p < 0.001.
    S1p Assay Kit, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Chemical Hypoxia Brings to Light Altered Autocrine Sphingosine-1-Phosphate Signalling in Rheumatoid Arthritis Synovial Fibroblasts"

    Article Title: Chemical Hypoxia Brings to Light Altered Autocrine Sphingosine-1-Phosphate Signalling in Rheumatoid Arthritis Synovial Fibroblasts

    Journal: Mediators of Inflammation

    doi: 10.1155/2015/436525

    Differential involvement of S1P receptors in S1P- and CoCl 2 -mediated chemokine secretion by normal FLS and RAFLS. Human primary FLS from normal ( n = 4) and RA ( n = 4) donors were incubated with 5 μ M S1P (a, b) or 200 μ M CoCl 2 (c, d). Where indicated, cells were pretreated with S1P 3 antagonist CAY10444 (5 μ M) or S1P 2 antagonist JTE-013 (5 μ M) for 30 min before stimulation with S1P or CoCl 2 . The amounts of chemokines released in the supernatants were monitored after 24 h. Data are expressed as percentage of chemokine production induced by S1P (a, b) or CoCl 2 (c, d). The data are the means ± SE from four experiments (4 different donors) performed in triplicate (3 independent experiments). For statistical comparative analyses, the samples stimulated with S1P (a, b) or CoCl 2 (c, d) were compared to those stimulated in the presence of CAY10444 or JTE-013, respectively. ∗∗ p < 0.01; ∗∗∗ p < 0.001.
    Figure Legend Snippet: Differential involvement of S1P receptors in S1P- and CoCl 2 -mediated chemokine secretion by normal FLS and RAFLS. Human primary FLS from normal ( n = 4) and RA ( n = 4) donors were incubated with 5 μ M S1P (a, b) or 200 μ M CoCl 2 (c, d). Where indicated, cells were pretreated with S1P 3 antagonist CAY10444 (5 μ M) or S1P 2 antagonist JTE-013 (5 μ M) for 30 min before stimulation with S1P or CoCl 2 . The amounts of chemokines released in the supernatants were monitored after 24 h. Data are expressed as percentage of chemokine production induced by S1P (a, b) or CoCl 2 (c, d). The data are the means ± SE from four experiments (4 different donors) performed in triplicate (3 independent experiments). For statistical comparative analyses, the samples stimulated with S1P (a, b) or CoCl 2 (c, d) were compared to those stimulated in the presence of CAY10444 or JTE-013, respectively. ∗∗ p < 0.01; ∗∗∗ p < 0.001.

    Techniques Used: Incubation

     S1P  content in normal FLS and RAFLS.
    Figure Legend Snippet: S1P content in normal FLS and RAFLS.

    Techniques Used:

    Impact of SPL inhibition on CoCl 2 -mediated chemokine/cytokine secretion in normal FLS and RAFLS. Human primary FLS from normal (a, c) and RA (b, d, e, f) donors were incubated with 200 μ M CoCl 2 in the presence of SPL inhibitor SM4 (3 μ M) or the inactive analog SM3 (3 μ M) and sphingosine (1 μ M) for 24 h. Where indicated, cells were pretreated with S1P 3 antagonist CAY10444 (5 μ M) or S1P 2 antagonist JTE-013 (5 μ M) for 30 min before stimulation with CoCl 2 in combination with sphingosine (Sph), SM4, or SM3. The data are the means ± SE from three experiments. For statistical comparative analyses, chemokine levels in the samples stimulated with CoCl 2 were compared to that of other samples (a–d) or chemokines produced by cells stimulated with CoCl 2 in combination with Sph and SM4 were compared to those produced by cells incubated with the S1P receptor antagonists prior to cell stimulation (f). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. Cytokine/chemokine secretion in RAFLS supernatants was analyzed using Proteome Profiler Human Cytokine Array panel A (e). Circled pairs of duplicate spots represent one cytokine/chemokine.
    Figure Legend Snippet: Impact of SPL inhibition on CoCl 2 -mediated chemokine/cytokine secretion in normal FLS and RAFLS. Human primary FLS from normal (a, c) and RA (b, d, e, f) donors were incubated with 200 μ M CoCl 2 in the presence of SPL inhibitor SM4 (3 μ M) or the inactive analog SM3 (3 μ M) and sphingosine (1 μ M) for 24 h. Where indicated, cells were pretreated with S1P 3 antagonist CAY10444 (5 μ M) or S1P 2 antagonist JTE-013 (5 μ M) for 30 min before stimulation with CoCl 2 in combination with sphingosine (Sph), SM4, or SM3. The data are the means ± SE from three experiments. For statistical comparative analyses, chemokine levels in the samples stimulated with CoCl 2 were compared to that of other samples (a–d) or chemokines produced by cells stimulated with CoCl 2 in combination with Sph and SM4 were compared to those produced by cells incubated with the S1P receptor antagonists prior to cell stimulation (f). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. Cytokine/chemokine secretion in RAFLS supernatants was analyzed using Proteome Profiler Human Cytokine Array panel A (e). Circled pairs of duplicate spots represent one cytokine/chemokine.

    Techniques Used: Inhibition, Incubation, Produced, Cell Stimulation

    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: 95/100, 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: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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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: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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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: 95/100, 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
    Immunosorbent Assay Elisa, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    sphingosine 1 phosphate assay kit  (Echelon Biosciences)


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    Echelon Biosciences sphingosine 1 phosphate assay kit
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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: 95/100, 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 s1p assay kit
    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) <t>S1P</t> was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.
    S1p Assay Kit, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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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).
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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).
    S1p Elisa Kit, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 95/100, 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).
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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).
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    Echelon Biosciences sphingosine 1 phosphate assay 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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    ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.

    Journal: PLoS ONE

    Article Title: Sphingosine Kinase-2 Maintains Viral Latency and Survival for KSHV-Infected Endothelial Cells

    doi: 10.1371/journal.pone.0102314

    Figure Lengend Snippet: ( A, B ) pDMVEC were incubated with or without purified KSHV for 2 h. After an additional 24 h, cells were incubated with the indicated concentrations of ABC294640 (ABC) or vehicle for another 24 h, then ceramide and dihydro-ceramide (dh-ceramide) species quantified as described in Methods. ( C, D ) Cells were treated as (A), then the concentrations of intracellular (C) or extracellular (D) S1P was quantified by ELISA. Error bars represent the S.E.M. for three independent experiments. * = p<0.01.

    Article Snippet: Concentrations of S1P in culture supernatants and pDMVEC cell lysates were determined using the S1P Assay Kit (Echelon) according to the manufacturers’ instructions.

    Techniques: Incubation, Purification, Enzyme-linked Immunosorbent Assay

    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