biotinylated  (Echelon Biosciences)


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    Echelon Biosciences biotinylated
    Biotinylated, 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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    biotinylated  (Echelon Biosciences)


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    Echelon Biosciences biotinylated
    Biotinylated, 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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    antibodies against ptdins 4 5 pe  (Echelon Biosciences)


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    Echelon Biosciences antibodies against ptdins 4 5 pe
    Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) <t>PtdIns(4,5)P2</t> and ( F ) PtdIns(3,4,5)P3 were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .
    Antibodies Against Ptdins 4 5 Pe, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 1 article reviews
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    1) Product Images from "Synergistic and additive interactions between receptor signaling networks drive the regulatory T cell versus T helper 17 cell fate choice"

    Article Title: Synergistic and additive interactions between receptor signaling networks drive the regulatory T cell versus T helper 17 cell fate choice

    Journal: The Journal of Biological Chemistry

    doi: 10.1016/j.jbc.2021.101330

    Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) PtdIns(4,5)P2 and ( F ) PtdIns(3,4,5)P3 were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .
    Figure Legend Snippet: Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) PtdIns(4,5)P2 and ( F ) PtdIns(3,4,5)P3 were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .

    Techniques Used: Isolation, Selection, Western Blot, Immunoprecipitation, Standard Deviation, Imaging, Flow Cytometry

    JAK2 inhibition dysregulates signaling networks during Th17 polarization. A , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, soluble anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-ZAP7(Y319) and total ZAP70. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Primary murine CD4 + T cells were activated under Th17 (TCR+TGF-β+IL-6) or Treg polarization conditions (TCR+TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. B , immunoblotting was performed for p-ZAP7(Y319), ZAP70, p-CSK-S364, and CSK. Densitometry was performed on immunoblots across three biological replicates from panel B to determine the relative amount of ( C ) p-CSK-S364 and ( D ) p-ZAP70(Y319) normalized to total CSK and Zap70 abundance respectively. Shown are mean ± SD; p values were calculated by one-way ANOVA. E , primary murine CD4 + T cells were activated under Th17 (TCR+ TGF-β+IL-6) or Treg polarization conditions (TCR+ TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. Immunoblotting was performed for p-P85 (Y458) and P85. F , densitometry was performed on immunoblots across three biological replicates from panel E were p-P85(Y458) was normalized to total P85. Shown are mean ± SD; p values were calculated by one-way ANOVA. Mass ELISA assays were utilized to determine the relative level of ( G ) PtdIns(3,4,5)P3, and ( H ) PtdIns(4,5)P2 generated in primary murine CD4 + T cells activated under Th17 or Treg polarization conditions for 10 min in the presence or absence of the JAK2 inhibitor. Shown are mean ± SD from three biological replicates; p values were calculated by one-way ANOVA. I , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-AKT(S473) and total AKT. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Source data are provided for the immunoblots in panels A , B , E , and I .
    Figure Legend Snippet: JAK2 inhibition dysregulates signaling networks during Th17 polarization. A , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, soluble anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-ZAP7(Y319) and total ZAP70. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Primary murine CD4 + T cells were activated under Th17 (TCR+TGF-β+IL-6) or Treg polarization conditions (TCR+TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. B , immunoblotting was performed for p-ZAP7(Y319), ZAP70, p-CSK-S364, and CSK. Densitometry was performed on immunoblots across three biological replicates from panel B to determine the relative amount of ( C ) p-CSK-S364 and ( D ) p-ZAP70(Y319) normalized to total CSK and Zap70 abundance respectively. Shown are mean ± SD; p values were calculated by one-way ANOVA. E , primary murine CD4 + T cells were activated under Th17 (TCR+ TGF-β+IL-6) or Treg polarization conditions (TCR+ TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. Immunoblotting was performed for p-P85 (Y458) and P85. F , densitometry was performed on immunoblots across three biological replicates from panel E were p-P85(Y458) was normalized to total P85. Shown are mean ± SD; p values were calculated by one-way ANOVA. Mass ELISA assays were utilized to determine the relative level of ( G ) PtdIns(3,4,5)P3, and ( H ) PtdIns(4,5)P2 generated in primary murine CD4 + T cells activated under Th17 or Treg polarization conditions for 10 min in the presence or absence of the JAK2 inhibitor. Shown are mean ± SD from three biological replicates; p values were calculated by one-way ANOVA. I , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-AKT(S473) and total AKT. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Source data are provided for the immunoblots in panels A , B , E , and I .

    Techniques Used: Inhibition, Western Blot, Enzyme-linked Immunosorbent Assay, Generated

    ptdins  (Echelon Biosciences)


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    Echelon Biosciences ptdins
    Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) PtdIns(4,5)P2 and ( F ) <t>PtdIns(3,4,5)P3</t> were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .
    Ptdins, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 1 article reviews
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    ptdins - by Bioz Stars, 2024-05
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    1) Product Images from "Synergistic and additive interactions between receptor signaling networks drive the regulatory T cell versus T helper 17 cell fate choice"

    Article Title: Synergistic and additive interactions between receptor signaling networks drive the regulatory T cell versus T helper 17 cell fate choice

    Journal: The Journal of Biological Chemistry

    doi: 10.1016/j.jbc.2021.101330

    Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) PtdIns(4,5)P2 and ( F ) PtdIns(3,4,5)P3 were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .
    Figure Legend Snippet: Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) PtdIns(4,5)P2 and ( F ) PtdIns(3,4,5)P3 were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .

    Techniques Used: Isolation, Selection, Western Blot, Immunoprecipitation, Standard Deviation, Imaging, Flow Cytometry

    JAK2 inhibition dysregulates signaling networks during Th17 polarization. A , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, soluble anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-ZAP7(Y319) and total ZAP70. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Primary murine CD4 + T cells were activated under Th17 (TCR+TGF-β+IL-6) or Treg polarization conditions (TCR+TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. B , immunoblotting was performed for p-ZAP7(Y319), ZAP70, p-CSK-S364, and CSK. Densitometry was performed on immunoblots across three biological replicates from panel B to determine the relative amount of ( C ) p-CSK-S364 and ( D ) p-ZAP70(Y319) normalized to total CSK and Zap70 abundance respectively. Shown are mean ± SD; p values were calculated by one-way ANOVA. E , primary murine CD4 + T cells were activated under Th17 (TCR+ TGF-β+IL-6) or Treg polarization conditions (TCR+ TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. Immunoblotting was performed for p-P85 (Y458) and P85. F , densitometry was performed on immunoblots across three biological replicates from panel E were p-P85(Y458) was normalized to total P85. Shown are mean ± SD; p values were calculated by one-way ANOVA. Mass ELISA assays were utilized to determine the relative level of ( G ) PtdIns(3,4,5)P3, and ( H ) PtdIns(4,5)P2 generated in primary murine CD4 + T cells activated under Th17 or Treg polarization conditions for 10 min in the presence or absence of the JAK2 inhibitor. Shown are mean ± SD from three biological replicates; p values were calculated by one-way ANOVA. I , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-AKT(S473) and total AKT. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Source data are provided for the immunoblots in panels A , B , E , and I .
    Figure Legend Snippet: JAK2 inhibition dysregulates signaling networks during Th17 polarization. A , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, soluble anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-ZAP7(Y319) and total ZAP70. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Primary murine CD4 + T cells were activated under Th17 (TCR+TGF-β+IL-6) or Treg polarization conditions (TCR+TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. B , immunoblotting was performed for p-ZAP7(Y319), ZAP70, p-CSK-S364, and CSK. Densitometry was performed on immunoblots across three biological replicates from panel B to determine the relative amount of ( C ) p-CSK-S364 and ( D ) p-ZAP70(Y319) normalized to total CSK and Zap70 abundance respectively. Shown are mean ± SD; p values were calculated by one-way ANOVA. E , primary murine CD4 + T cells were activated under Th17 (TCR+ TGF-β+IL-6) or Treg polarization conditions (TCR+ TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. Immunoblotting was performed for p-P85 (Y458) and P85. F , densitometry was performed on immunoblots across three biological replicates from panel E were p-P85(Y458) was normalized to total P85. Shown are mean ± SD; p values were calculated by one-way ANOVA. Mass ELISA assays were utilized to determine the relative level of ( G ) PtdIns(3,4,5)P3, and ( H ) PtdIns(4,5)P2 generated in primary murine CD4 + T cells activated under Th17 or Treg polarization conditions for 10 min in the presence or absence of the JAK2 inhibitor. Shown are mean ± SD from three biological replicates; p values were calculated by one-way ANOVA. I , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-AKT(S473) and total AKT. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Source data are provided for the immunoblots in panels A , B , E , and I .

    Techniques Used: Inhibition, Western Blot, Enzyme-linked Immunosorbent Assay, Generated

    antibodies against pi 4 5 p2 pe  (Echelon Biosciences)


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    Echelon Biosciences antibodies against pi 4 5 p2 pe
    Antibodies Against Pi 4 5 P2 Pe, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 1 article reviews
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    antibodies against pi 4 5 p2 pe - by Bioz Stars, 2024-05
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    pip3 pe  (Echelon Biosciences)


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    Echelon Biosciences pip3 pe
    Pip3 Pe, 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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    z-b045  (Echelon Biosciences)


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    Echelon Biosciences z-b045
    Z B045, 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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    mouse echelon biosciences pip 2 z b045  (Echelon Biosciences)


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    Echelon Biosciences mouse echelon biosciences pip 2 z b045
    Primary antibodies used in this study
    Mouse Echelon Biosciences Pip 2 Z B045, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Choroid plexus epithelial cells express the adhesion protein P-cadherin at cell-cell contacts and syntaxin-4 in the luminal membrane domain"

    Article Title: Choroid plexus epithelial cells express the adhesion protein P-cadherin at cell-cell contacts and syntaxin-4 in the luminal membrane domain

    Journal: American Journal of Physiology - Cell Physiology

    doi: 10.1152/ajpcell.00305.2017

    Primary antibodies used in this study
    Figure Legend Snippet: Primary antibodies used in this study

    Techniques Used:

    mouse echelon biosciences pip 2 z b045  (Echelon Biosciences)


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    Echelon Biosciences mouse echelon biosciences pip 2 z b045
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    Mouse Echelon Biosciences Pip 2 Z B045, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Choroid plexus epithelial cells express the adhesion protein P-cadherin at cell-cell contacts and syntaxin-4 in the luminal membrane domain"

    Article Title: Choroid plexus epithelial cells express the adhesion protein P-cadherin at cell-cell contacts and syntaxin-4 in the luminal membrane domain

    Journal: American Journal of Physiology - Cell Physiology

    doi: 10.1152/ajpcell.00305.2017

    Primary antibodies used in this study
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    z b045  (Echelon Biosciences)


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    Z B045, 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 "Choroid plexus epithelial cells express the adhesion protein P-cadherin at cell-cell contacts and syntaxin-4 in the luminal membrane domain"

    Article Title: Choroid plexus epithelial cells express the adhesion protein P-cadherin at cell-cell contacts and syntaxin-4 in the luminal membrane domain

    Journal: American Journal of Physiology - Cell Physiology

    doi: 10.1152/ajpcell.00305.2017

    Primary antibodies used in this study
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    mouse echelon biosciences pip 2 z b045  (Echelon Biosciences)


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    Echelon Biosciences mouse echelon biosciences pip 2 z b045
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    Mouse Echelon Biosciences Pip 2 Z B045, 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 "Choroid plexus epithelial cells express the adhesion protein P-cadherin at cell-cell contacts and syntaxin-4 in the luminal membrane domain"

    Article Title: Choroid plexus epithelial cells express the adhesion protein P-cadherin at cell-cell contacts and syntaxin-4 in the luminal membrane domain

    Journal: American Journal of Physiology - Cell Physiology

    doi: 10.1152/ajpcell.00305.2017

    Primary antibodies used in this study
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    Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) <t>PtdIns(4,5)P2</t> and ( F ) PtdIns(3,4,5)P3 were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .
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    Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) PtdIns(4,5)P2 and ( F ) <t>PtdIns(3,4,5)P3</t> were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .
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    Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) PtdIns(4,5)P2 and ( F ) <t>PtdIns(3,4,5)P3</t> were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .
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    Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) PtdIns(4,5)P2 and ( F ) <t>PtdIns(3,4,5)P3</t> were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .
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    Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) PtdIns(4,5)P2 and ( F ) <t>PtdIns(3,4,5)P3</t> were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .
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    Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) PtdIns(4,5)P2 and ( F ) PtdIns(3,4,5)P3 were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .

    Journal: The Journal of Biological Chemistry

    Article Title: Synergistic and additive interactions between receptor signaling networks drive the regulatory T cell versus T helper 17 cell fate choice

    doi: 10.1016/j.jbc.2021.101330

    Figure Lengend Snippet: Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) PtdIns(4,5)P2 and ( F ) PtdIns(3,4,5)P3 were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .

    Article Snippet: Cells were also stained with antibodies against PtdIns(4,5)-PE (Echelon Biosciences Z-B045) or PtdIns(3,4,5)P3-PE (Echelon Biosciences Z-B3345B).

    Techniques: Isolation, Selection, Western Blot, Immunoprecipitation, Standard Deviation, Imaging, Flow Cytometry

    JAK2 inhibition dysregulates signaling networks during Th17 polarization. A , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, soluble anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-ZAP7(Y319) and total ZAP70. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Primary murine CD4 + T cells were activated under Th17 (TCR+TGF-β+IL-6) or Treg polarization conditions (TCR+TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. B , immunoblotting was performed for p-ZAP7(Y319), ZAP70, p-CSK-S364, and CSK. Densitometry was performed on immunoblots across three biological replicates from panel B to determine the relative amount of ( C ) p-CSK-S364 and ( D ) p-ZAP70(Y319) normalized to total CSK and Zap70 abundance respectively. Shown are mean ± SD; p values were calculated by one-way ANOVA. E , primary murine CD4 + T cells were activated under Th17 (TCR+ TGF-β+IL-6) or Treg polarization conditions (TCR+ TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. Immunoblotting was performed for p-P85 (Y458) and P85. F , densitometry was performed on immunoblots across three biological replicates from panel E were p-P85(Y458) was normalized to total P85. Shown are mean ± SD; p values were calculated by one-way ANOVA. Mass ELISA assays were utilized to determine the relative level of ( G ) PtdIns(3,4,5)P3, and ( H ) PtdIns(4,5)P2 generated in primary murine CD4 + T cells activated under Th17 or Treg polarization conditions for 10 min in the presence or absence of the JAK2 inhibitor. Shown are mean ± SD from three biological replicates; p values were calculated by one-way ANOVA. I , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-AKT(S473) and total AKT. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Source data are provided for the immunoblots in panels A , B , E , and I .

    Journal: The Journal of Biological Chemistry

    Article Title: Synergistic and additive interactions between receptor signaling networks drive the regulatory T cell versus T helper 17 cell fate choice

    doi: 10.1016/j.jbc.2021.101330

    Figure Lengend Snippet: JAK2 inhibition dysregulates signaling networks during Th17 polarization. A , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, soluble anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-ZAP7(Y319) and total ZAP70. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Primary murine CD4 + T cells were activated under Th17 (TCR+TGF-β+IL-6) or Treg polarization conditions (TCR+TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. B , immunoblotting was performed for p-ZAP7(Y319), ZAP70, p-CSK-S364, and CSK. Densitometry was performed on immunoblots across three biological replicates from panel B to determine the relative amount of ( C ) p-CSK-S364 and ( D ) p-ZAP70(Y319) normalized to total CSK and Zap70 abundance respectively. Shown are mean ± SD; p values were calculated by one-way ANOVA. E , primary murine CD4 + T cells were activated under Th17 (TCR+ TGF-β+IL-6) or Treg polarization conditions (TCR+ TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. Immunoblotting was performed for p-P85 (Y458) and P85. F , densitometry was performed on immunoblots across three biological replicates from panel E were p-P85(Y458) was normalized to total P85. Shown are mean ± SD; p values were calculated by one-way ANOVA. Mass ELISA assays were utilized to determine the relative level of ( G ) PtdIns(3,4,5)P3, and ( H ) PtdIns(4,5)P2 generated in primary murine CD4 + T cells activated under Th17 or Treg polarization conditions for 10 min in the presence or absence of the JAK2 inhibitor. Shown are mean ± SD from three biological replicates; p values were calculated by one-way ANOVA. I , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-AKT(S473) and total AKT. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Source data are provided for the immunoblots in panels A , B , E , and I .

    Article Snippet: Cells were also stained with antibodies against PtdIns(4,5)-PE (Echelon Biosciences Z-B045) or PtdIns(3,4,5)P3-PE (Echelon Biosciences Z-B3345B).

    Techniques: Inhibition, Western Blot, Enzyme-linked Immunosorbent Assay, Generated

    Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) PtdIns(4,5)P2 and ( F ) PtdIns(3,4,5)P3 were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .

    Journal: The Journal of Biological Chemistry

    Article Title: Synergistic and additive interactions between receptor signaling networks drive the regulatory T cell versus T helper 17 cell fate choice

    doi: 10.1016/j.jbc.2021.101330

    Figure Lengend Snippet: Phosphatidylinositol metabolism is differentially regulated during Treg versus Th17 induction. A , a schematic of phosphatidylinositol metabolism is depicted. B , the label-free phosphoproteomic analysis determined the relative abundance of phosphopeptides for phosphatidylinositol kinases and phosphatases regulated by signaling input. C , primary CD4 + murine T cells isolated by negative selection were activated under Treg (anti-CD3 AB, soluble CD28 AB, and TGF-β) or Th17 (anti-CD3 AB, soluble anti-CD28 AB, TGF-β, and IL-6) induction conditions. Immunoblotting was performed for p-P85 (Y458) and total P85. Densitometry was performed across three biological replicates to quantitate p-P85 (Y458) normalized to total p85 levels. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01). D , primary murine CD4 + T cells were activated with under TH0 (anti-CD3, soluble anti-CD28 antibodies), Treg (anti-CD3 and soluble anti-CD28 antibody and TGF-β) and Th17 (anti-CD3 antibody, anti-CD28 antibody, TGF-β, and IL-6) for 10 min. P85 was immunoprecipitated. Immunoblotting was performed for P85 and P110. Densitometry was performed across three biological replicates. Each data point represents the mean ± standard deviation across three independent experiments and one-way ANOVA was performed. The abundances of ( E ) PtdIns(4,5)P2 and ( F ) PtdIns(3,4,5)P3 were measured with an imaging flow cytometry assay in primary murine CD4 + T cells activated under TH0, Treg, and Th17 conditions (defined above). Three biological replicates were performed. Presented is the mean ± standard deviation. G , primary murine CD4 + T cells were activated under Treg and Th17 conditions (defined above), and immunoblotting was performed on the resulting lysates for p-AKT(S473) and total AKT. Densitometry was performed on the immunoblots. Three biological replicates were included. Phosphorylated AKT was normalized to the total AKT abundance. Shown are mean ± SD; p values were calculated by two-way ANOVA (∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001). Source data are provided for the immunoblots in panels C , D , and G .

    Article Snippet: Cells were also stained with antibodies against PtdIns(4,5)-PE (Echelon Biosciences Z-B045) or PtdIns(3,4,5)P3-PE (Echelon Biosciences Z-B3345B).

    Techniques: Isolation, Selection, Western Blot, Immunoprecipitation, Standard Deviation, Imaging, Flow Cytometry

    JAK2 inhibition dysregulates signaling networks during Th17 polarization. A , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, soluble anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-ZAP7(Y319) and total ZAP70. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Primary murine CD4 + T cells were activated under Th17 (TCR+TGF-β+IL-6) or Treg polarization conditions (TCR+TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. B , immunoblotting was performed for p-ZAP7(Y319), ZAP70, p-CSK-S364, and CSK. Densitometry was performed on immunoblots across three biological replicates from panel B to determine the relative amount of ( C ) p-CSK-S364 and ( D ) p-ZAP70(Y319) normalized to total CSK and Zap70 abundance respectively. Shown are mean ± SD; p values were calculated by one-way ANOVA. E , primary murine CD4 + T cells were activated under Th17 (TCR+ TGF-β+IL-6) or Treg polarization conditions (TCR+ TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. Immunoblotting was performed for p-P85 (Y458) and P85. F , densitometry was performed on immunoblots across three biological replicates from panel E were p-P85(Y458) was normalized to total P85. Shown are mean ± SD; p values were calculated by one-way ANOVA. Mass ELISA assays were utilized to determine the relative level of ( G ) PtdIns(3,4,5)P3, and ( H ) PtdIns(4,5)P2 generated in primary murine CD4 + T cells activated under Th17 or Treg polarization conditions for 10 min in the presence or absence of the JAK2 inhibitor. Shown are mean ± SD from three biological replicates; p values were calculated by one-way ANOVA. I , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-AKT(S473) and total AKT. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Source data are provided for the immunoblots in panels A , B , E , and I .

    Journal: The Journal of Biological Chemistry

    Article Title: Synergistic and additive interactions between receptor signaling networks drive the regulatory T cell versus T helper 17 cell fate choice

    doi: 10.1016/j.jbc.2021.101330

    Figure Lengend Snippet: JAK2 inhibition dysregulates signaling networks during Th17 polarization. A , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, soluble anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-ZAP7(Y319) and total ZAP70. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Primary murine CD4 + T cells were activated under Th17 (TCR+TGF-β+IL-6) or Treg polarization conditions (TCR+TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. B , immunoblotting was performed for p-ZAP7(Y319), ZAP70, p-CSK-S364, and CSK. Densitometry was performed on immunoblots across three biological replicates from panel B to determine the relative amount of ( C ) p-CSK-S364 and ( D ) p-ZAP70(Y319) normalized to total CSK and Zap70 abundance respectively. Shown are mean ± SD; p values were calculated by one-way ANOVA. E , primary murine CD4 + T cells were activated under Th17 (TCR+ TGF-β+IL-6) or Treg polarization conditions (TCR+ TGF-β) for 10 min in the presence or absence of 200 nM JAK2 inhibitor. Immunoblotting was performed for p-P85 (Y458) and P85. F , densitometry was performed on immunoblots across three biological replicates from panel E were p-P85(Y458) was normalized to total P85. Shown are mean ± SD; p values were calculated by one-way ANOVA. Mass ELISA assays were utilized to determine the relative level of ( G ) PtdIns(3,4,5)P3, and ( H ) PtdIns(4,5)P2 generated in primary murine CD4 + T cells activated under Th17 or Treg polarization conditions for 10 min in the presence or absence of the JAK2 inhibitor. Shown are mean ± SD from three biological replicates; p values were calculated by one-way ANOVA. I , primary murine CD4 + T cells were activated for 10 min with constant amounts of anti-CD3 antibody, anti-CD28 antibody, and TGF-β with varying amounts of IL-6. Immunoblotting was performed for p-AKT(S473) and total AKT. Densitometry was performed on immunoblots across three biological replicates. Shown are mean ± SD. Source data are provided for the immunoblots in panels A , B , E , and I .

    Article Snippet: Cells were also stained with antibodies against PtdIns(4,5)-PE (Echelon Biosciences Z-B045) or PtdIns(3,4,5)P3-PE (Echelon Biosciences Z-B3345B).

    Techniques: Inhibition, Western Blot, Enzyme-linked Immunosorbent Assay, Generated

    Primary antibodies used in this study

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Choroid plexus epithelial cells express the adhesion protein P-cadherin at cell-cell contacts and syntaxin-4 in the luminal membrane domain

    doi: 10.1152/ajpcell.00305.2017

    Figure Lengend Snippet: Primary antibodies used in this study

    Article Snippet: PIP 3 Z-P345b Floating 1:100 Mouse Echelon Biosciences PIP 2 Z-B045 Floating 1:100 Mouse Echelon Biosciences Pals1 17710-1-AP Paraffin–TEG 1:100 Rabbit Proteintech Crumbs 3 crumbs 3A Paraffin–TEG 1:200 Rat Massey-Harroche, Le Bivic PKCζ sc216 Paraffin–TEG 1:500 Rabbit Santa Cruz Biotech.

    Techniques:

    Primary antibodies used in this study

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Choroid plexus epithelial cells express the adhesion protein P-cadherin at cell-cell contacts and syntaxin-4 in the luminal membrane domain

    doi: 10.1152/ajpcell.00305.2017

    Figure Lengend Snippet: Primary antibodies used in this study

    Article Snippet: PIP 2 , Z-B045 , Floating , 1:100 , Mouse , Echelon Biosciences.

    Techniques: