pathscan phospho mek1  (Cell Signaling Technology Inc)

 
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    Name:
    PathScan Phospho MEK1 Ser217 221 Sandwich ELISA Kit
    Description:
    CST s PathScan Phospho MEK1 Ser217 221 Sandwich ELISA Kit is a solid phase sandwich enzyme linked immunosorbent assay ELISA that detects endogenous levels of Phospho MEK1 Ser217 221 protein MEK1 Mouse mAb has been coated onto the microwells After incubation with cell lysates total MEK1 protein phospho and nonphospho is captured by the coated antibody Following extensive washing a Phospho MEK1 2 Antibody is added to detect the captured phospho MEK1 protein Anti rabbit IgG HRP linked antibody is then used to recognize the bound detection antibody HRP substrate TMB is added to develop color The magnitude of optical density for this developed color is proportional to the quantity of phospho MEK1 protein
    Catalog Number:
    7175
    Price:
    None
    Category:
    ELISA Kits
    Reactivity:
    Human Mouse
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    Structured Review

    Cell Signaling Technology Inc pathscan phospho mek1
    CST s PathScan Phospho MEK1 Ser217 221 Sandwich ELISA Kit is a solid phase sandwich enzyme linked immunosorbent assay ELISA that detects endogenous levels of Phospho MEK1 Ser217 221 protein MEK1 Mouse mAb has been coated onto the microwells After incubation with cell lysates total MEK1 protein phospho and nonphospho is captured by the coated antibody Following extensive washing a Phospho MEK1 2 Antibody is added to detect the captured phospho MEK1 protein Anti rabbit IgG HRP linked antibody is then used to recognize the bound detection antibody HRP substrate TMB is added to develop color The magnitude of optical density for this developed color is proportional to the quantity of phospho MEK1 protein
    https://www.bioz.com/result/pathscan phospho mek1/product/Cell Signaling Technology Inc
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pathscan phospho mek1 - by Bioz Stars, 2020-09
    90/100 stars

    Related Products / Commonly Used Together

    total mek1 sandwich elisa kits

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    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: Achillolide A Protects Astrocytes against Oxidative Stress by Reducing Intracellular Reactive Oxygen Species and Interfering with Cell Signaling
    Article Snippet: .. To measure the amount of total and phosphorylated MEK1 in cell lysates of astrocytes, ELISA was performed according to the manufacturer’s protocol using the PathScan total MEK1 sandwich ELISA kit (Cell Signaling Technology) and the PathScan phosoho-MEK1 (Ser217/221) sandwich ELISA kit (Cell Signaling Technology), respectively. .. To measure the amount of total and phospho-p44/42 MAPK in cell lysates of astrocytes, ELISA was performed according to the manufacturer’s protocol using the PathScan phosoho-p44/42 MAPK (Thr202/Tyr204) sandwich ELISA kit (Cell Signaling Technology), or the PathScan total p44/42 MAPK sandwich ELISA kit (Cell Signaling Technology), respectively.

    Article Title: Huntingtin Subcellular Localisation Is Regulated by Kinase Signalling Activity in the StHdhQ111 Model of HD
    Article Snippet: .. ELISA Phospho-AKT1 (Ser473) and phospho-MEK1 (Ser217/221) PathScan® Sandwich ELISA kits (Cell Signalling Technology) were utilised to assay kinase activation and inhibition, and were carried out according to manufacturer’s instructions. .. Kinase activation and inhibition and huntingtin localisation EGF (Life Technologies) was reconstituted and diluted to a stock solution of 100μg/ml in PBS and stored at -20°C.

    Sandwich ELISA:

    Article Title: Achillolide A Protects Astrocytes against Oxidative Stress by Reducing Intracellular Reactive Oxygen Species and Interfering with Cell Signaling
    Article Snippet: .. To measure the amount of total and phosphorylated MEK1 in cell lysates of astrocytes, ELISA was performed according to the manufacturer’s protocol using the PathScan total MEK1 sandwich ELISA kit (Cell Signaling Technology) and the PathScan phosoho-MEK1 (Ser217/221) sandwich ELISA kit (Cell Signaling Technology), respectively. .. To measure the amount of total and phospho-p44/42 MAPK in cell lysates of astrocytes, ELISA was performed according to the manufacturer’s protocol using the PathScan phosoho-p44/42 MAPK (Thr202/Tyr204) sandwich ELISA kit (Cell Signaling Technology), or the PathScan total p44/42 MAPK sandwich ELISA kit (Cell Signaling Technology), respectively.

    Article Title: Huntingtin Subcellular Localisation Is Regulated by Kinase Signalling Activity in the StHdhQ111 Model of HD
    Article Snippet: .. ELISA Phospho-AKT1 (Ser473) and phospho-MEK1 (Ser217/221) PathScan® Sandwich ELISA kits (Cell Signalling Technology) were utilised to assay kinase activation and inhibition, and were carried out according to manufacturer’s instructions. .. Kinase activation and inhibition and huntingtin localisation EGF (Life Technologies) was reconstituted and diluted to a stock solution of 100μg/ml in PBS and stored at -20°C.

    Article Title: An Evolutionary Conserved Role for Anaplastic Lymphoma Kinase in Behavioral Responses to Ethanol
    Article Snippet: .. Striatum was dissected from the brains of adult male mice and analyzed using the PathScan® Phospho-MEK1 and Total MEK1 Sandwich ELISA kits from Cell Signaling Technology. .. Shown is the ratio of phosphorylated MEK to total MEK (pMEK/MEK) in wild-type (+/+) and AlkKO (−/−) mice.

    Inhibition:

    Article Title: Huntingtin Subcellular Localisation Is Regulated by Kinase Signalling Activity in the StHdhQ111 Model of HD
    Article Snippet: .. ELISA Phospho-AKT1 (Ser473) and phospho-MEK1 (Ser217/221) PathScan® Sandwich ELISA kits (Cell Signalling Technology) were utilised to assay kinase activation and inhibition, and were carried out according to manufacturer’s instructions. .. Kinase activation and inhibition and huntingtin localisation EGF (Life Technologies) was reconstituted and diluted to a stock solution of 100μg/ml in PBS and stored at -20°C.

    Activation Assay:

    Article Title: Huntingtin Subcellular Localisation Is Regulated by Kinase Signalling Activity in the StHdhQ111 Model of HD
    Article Snippet: .. ELISA Phospho-AKT1 (Ser473) and phospho-MEK1 (Ser217/221) PathScan® Sandwich ELISA kits (Cell Signalling Technology) were utilised to assay kinase activation and inhibition, and were carried out according to manufacturer’s instructions. .. Kinase activation and inhibition and huntingtin localisation EGF (Life Technologies) was reconstituted and diluted to a stock solution of 100μg/ml in PBS and stored at -20°C.

    Mouse Assay:

    Article Title: An Evolutionary Conserved Role for Anaplastic Lymphoma Kinase in Behavioral Responses to Ethanol
    Article Snippet: .. Striatum was dissected from the brains of adult male mice and analyzed using the PathScan® Phospho-MEK1 and Total MEK1 Sandwich ELISA kits from Cell Signaling Technology. .. Shown is the ratio of phosphorylated MEK to total MEK (pMEK/MEK) in wild-type (+/+) and AlkKO (−/−) mice.

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    Cell Signaling Technology Inc phospho mek1 2
    MKP-3 knockdown via siRNA increases ERK1/2 phosphorylation in H- ras MCF10A and DLD-1 cells. (A) Whole cell lysates were prepared from serum starved MCF10A and H- ras MCF10A cells. MKP-3 ( top panel ) was detected by immunoblot analysis. The blot was stripped and reprobed for β-tubulin ( bottom panel ) as a loading control. Whole cell lysates were prepared from (B) H- ras MCF10A cells or (C) DLD-1 cells that were either not transfected (lane 1), were transfected with duplex siRNA targeted against MKP-3 (lane 2), or were transfected with duplex scrambled siRNA (lane 3). The following proteins were detected by immunoblot analysis: dually phosphorylated, active <t>MEK1/2</t> (pMEK1/2); total MEK1; dually phosphorylated, active ERK1/2 (pERK1/2); total ERK2; or MKP-3. The MKP-3 blot was stripped and reprobed for β-tubulin as a loading control. As a positive control for phospho-MEK1/2, nontransfected cells were incubated for 120 minutes with (B) 1.6 nM TPA (lane 4), or incubated for 30 minutes with (C) 3 nM EGF (lane 4). ). The graphs represent average pERK1/2 densitometry relative to non-transfected controls for (B) 10 independent experiments and (C) 6 independent experiments; error bars represent SEM. Filled bars (nt), not transfected. Open bars (M), transfected with duplex siRNA targeted against MKP-3. Hatched bars (S), transfected with duplex scrambled siRNA. Treatment of H- ras MCF10A and DLD-1 cells with MKP3 siRNA resulted in an increase in pERK1/2 levels relative to nontransfected control or treatment with scrambled siRNA (H- ras MCF10A: p = 0.006, n=30, 1-way ANOVA and Tukey’s HSD, α=0.05; DLD-1: p= 0.0008, n=18, 1-way ANOVA and Tukey’s HSD, α=0.05). We did not detect a difference between non transfected cells or cells transfected with scrambled siRNA.
    Phospho Mek1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho mek1 2/product/Cell Signaling Technology Inc
    Average 99 stars, based on 10 article reviews
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    91
    Cell Signaling Technology Inc pathscan multitarget elisa
    <t>PathScan</t> MAPK <t>multitarget</t> sandwich <t>ELISA.</t> HOEC monolayers (mixed donors; N = 3) were treated with either TNF-α (10 ng/ml), IL-1β (10 ng/ml), hBD-2 (5 μg/ml), or hBD-3 (5 μg/ml) for 0, 30, or 60 min, respectively. The cell
    Pathscan Multitarget Elisa, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pathscan multitarget elisa/product/Cell Signaling Technology Inc
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pathscan multitarget elisa - by Bioz Stars, 2020-09
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    90
    Cell Signaling Technology Inc pathscan total mek1 sandwich elisa kit
    Inhibitory effect of achillolide A on H 2 O 2 -induced phosphorylation of p44/42 MAPK and <t>MEK1</t> in astrocytes. Astrocytes were treated with 175 μM of H 2 O 2 for 40 min following preincubation with achillolide A or memantine for 2 h. The levels of phosphorylated and total p44/42 MAPK ( A , B ) and phosphorylated and total MEK1 ( C ) were measured by <t>ELISA.</t> Relative phospho MAPK levels were calculated from the ratio: phospho MAPK levels/total MAPK levels the results are means ± SEM of two experiments ( n = 4). * p
    Pathscan Total Mek1 Sandwich Elisa Kit, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pathscan total mek1 sandwich elisa kit/product/Cell Signaling Technology Inc
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pathscan total mek1 sandwich elisa kit - by Bioz Stars, 2020-09
    90/100 stars
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    94
    Cell Signaling Technology Inc pathscan multi target sandwich elisa kit
    SPI − inhibits LPS-induced NF-κB activation. ApoE–/– mice ( n = 4/diet) were fed CAS- or SPI − -diet for 1 week followed by LPS challenge for 3 h. Nuclear and cytosolic fractions were prepared from aorta and liver lysates (experiment 5). Total- and phospho-NF-κB in nuclear fractions of aorta ( a ) and liver ( b ) were determined by NF-κB <t>Pathscan</t> <t>ELISA</t> kit. Cytosolic I-κB protein levels ( c , d ) in aortic lysates from CAS- or SPI − -fed apoE–/– mice were determined by Western blot. Blots probed with GAPDH were used to normalize I-kB protein levels. Values are mean ± SD, n = 3. Immunofluorescence analysis of NF-κB p65 nuclear translocation in liver CAS- or SPI − -fed mice ( n = 4). Representative pictures from 2 mice per diet group are represented. Liver sections (10 μm) were fixed in PBS/10 % formalin and stained with rabbit anti-NF-κb p65 mAb as described under section “Materials and methods.” Sections were mounted with DAPI to localize nucleus. Number of nucleated (DAPI+) and NF-κB-positive cells (f) were counted in 3 separate sections/mouse. ** P
    Pathscan Multi Target Sandwich Elisa Kit, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pathscan multi target sandwich elisa kit/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
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    Image Search Results


    MKP-3 knockdown via siRNA increases ERK1/2 phosphorylation in H- ras MCF10A and DLD-1 cells. (A) Whole cell lysates were prepared from serum starved MCF10A and H- ras MCF10A cells. MKP-3 ( top panel ) was detected by immunoblot analysis. The blot was stripped and reprobed for β-tubulin ( bottom panel ) as a loading control. Whole cell lysates were prepared from (B) H- ras MCF10A cells or (C) DLD-1 cells that were either not transfected (lane 1), were transfected with duplex siRNA targeted against MKP-3 (lane 2), or were transfected with duplex scrambled siRNA (lane 3). The following proteins were detected by immunoblot analysis: dually phosphorylated, active MEK1/2 (pMEK1/2); total MEK1; dually phosphorylated, active ERK1/2 (pERK1/2); total ERK2; or MKP-3. The MKP-3 blot was stripped and reprobed for β-tubulin as a loading control. As a positive control for phospho-MEK1/2, nontransfected cells were incubated for 120 minutes with (B) 1.6 nM TPA (lane 4), or incubated for 30 minutes with (C) 3 nM EGF (lane 4). ). The graphs represent average pERK1/2 densitometry relative to non-transfected controls for (B) 10 independent experiments and (C) 6 independent experiments; error bars represent SEM. Filled bars (nt), not transfected. Open bars (M), transfected with duplex siRNA targeted against MKP-3. Hatched bars (S), transfected with duplex scrambled siRNA. Treatment of H- ras MCF10A and DLD-1 cells with MKP3 siRNA resulted in an increase in pERK1/2 levels relative to nontransfected control or treatment with scrambled siRNA (H- ras MCF10A: p = 0.006, n=30, 1-way ANOVA and Tukey’s HSD, α=0.05; DLD-1: p= 0.0008, n=18, 1-way ANOVA and Tukey’s HSD, α=0.05). We did not detect a difference between non transfected cells or cells transfected with scrambled siRNA.

    Journal: Toxicology and applied pharmacology

    Article Title: Reciprocal Regulation of Extracellular Signal Regulated Kinase 1/2 and Mitogen Activated Protein Kinase Phosphatase-3

    doi: 10.1016/j.taap.2008.08.007

    Figure Lengend Snippet: MKP-3 knockdown via siRNA increases ERK1/2 phosphorylation in H- ras MCF10A and DLD-1 cells. (A) Whole cell lysates were prepared from serum starved MCF10A and H- ras MCF10A cells. MKP-3 ( top panel ) was detected by immunoblot analysis. The blot was stripped and reprobed for β-tubulin ( bottom panel ) as a loading control. Whole cell lysates were prepared from (B) H- ras MCF10A cells or (C) DLD-1 cells that were either not transfected (lane 1), were transfected with duplex siRNA targeted against MKP-3 (lane 2), or were transfected with duplex scrambled siRNA (lane 3). The following proteins were detected by immunoblot analysis: dually phosphorylated, active MEK1/2 (pMEK1/2); total MEK1; dually phosphorylated, active ERK1/2 (pERK1/2); total ERK2; or MKP-3. The MKP-3 blot was stripped and reprobed for β-tubulin as a loading control. As a positive control for phospho-MEK1/2, nontransfected cells were incubated for 120 minutes with (B) 1.6 nM TPA (lane 4), or incubated for 30 minutes with (C) 3 nM EGF (lane 4). ). The graphs represent average pERK1/2 densitometry relative to non-transfected controls for (B) 10 independent experiments and (C) 6 independent experiments; error bars represent SEM. Filled bars (nt), not transfected. Open bars (M), transfected with duplex siRNA targeted against MKP-3. Hatched bars (S), transfected with duplex scrambled siRNA. Treatment of H- ras MCF10A and DLD-1 cells with MKP3 siRNA resulted in an increase in pERK1/2 levels relative to nontransfected control or treatment with scrambled siRNA (H- ras MCF10A: p = 0.006, n=30, 1-way ANOVA and Tukey’s HSD, α=0.05; DLD-1: p= 0.0008, n=18, 1-way ANOVA and Tukey’s HSD, α=0.05). We did not detect a difference between non transfected cells or cells transfected with scrambled siRNA.

    Article Snippet: After blocking in a TBST/5% milk solution, immunoblots were incubated overnight at 4°C using the following primary antibodies and dilutions: Phospho-p44/42 MAPK (Thr-202/Tyr-204) (E10) (mouse monoclonal) (1:2000), and phospho-MEK1/2 (Ser-217/221) (rabbit polyclonal) (1:2000) from Cell Signaling (Beverly, MA), and ERK2 (C-14) (rabbit polyclonal) (1:2000), MEK1 (12-B) (rabbit polyclonal) (1:500), MKP-3 (C-20) (goat polyclonal) (1:1000) and β-tubulin (H-235) (rabbit polyclonal) (1:2000) from Santa Cruz Biotechnology (Santa Cruz, CA).

    Techniques: Transfection, Positive Control, Incubation

    TPA and EGF stimulate the loss and recovery of MKP-3 protein. Whole cell lysates were prepared from (A) H- ras MCF10A cells that were incubated for the indicated times with 1.6 nM TPA; (B) H- ras MCF10A cells; or (C) DLD-1 cells that were incubated for the indicated times with 3 nM EGF. The following proteins were detected by immunoblot analysis: dually phosphorylated, active MEK1/2 (pMEK1/2); total MEK1; dually phosphorylated, active ERK1/2 (pERK1/2); total ERK2; or MKP-3. The MKP-3 blot was stripped and reprobed for β-tubulin as a loading control. The graphs represent average densitometry of pMEK1/2, pERK1/2, and MKP-3 for 3 independent experiments; error bars represent SEM. Filled bars, pMEK1/2. Open bars, pERK1/2. Hatched bars, MKP-3.

    Journal: Toxicology and applied pharmacology

    Article Title: Reciprocal Regulation of Extracellular Signal Regulated Kinase 1/2 and Mitogen Activated Protein Kinase Phosphatase-3

    doi: 10.1016/j.taap.2008.08.007

    Figure Lengend Snippet: TPA and EGF stimulate the loss and recovery of MKP-3 protein. Whole cell lysates were prepared from (A) H- ras MCF10A cells that were incubated for the indicated times with 1.6 nM TPA; (B) H- ras MCF10A cells; or (C) DLD-1 cells that were incubated for the indicated times with 3 nM EGF. The following proteins were detected by immunoblot analysis: dually phosphorylated, active MEK1/2 (pMEK1/2); total MEK1; dually phosphorylated, active ERK1/2 (pERK1/2); total ERK2; or MKP-3. The MKP-3 blot was stripped and reprobed for β-tubulin as a loading control. The graphs represent average densitometry of pMEK1/2, pERK1/2, and MKP-3 for 3 independent experiments; error bars represent SEM. Filled bars, pMEK1/2. Open bars, pERK1/2. Hatched bars, MKP-3.

    Article Snippet: After blocking in a TBST/5% milk solution, immunoblots were incubated overnight at 4°C using the following primary antibodies and dilutions: Phospho-p44/42 MAPK (Thr-202/Tyr-204) (E10) (mouse monoclonal) (1:2000), and phospho-MEK1/2 (Ser-217/221) (rabbit polyclonal) (1:2000) from Cell Signaling (Beverly, MA), and ERK2 (C-14) (rabbit polyclonal) (1:2000), MEK1 (12-B) (rabbit polyclonal) (1:500), MKP-3 (C-20) (goat polyclonal) (1:1000) and β-tubulin (H-235) (rabbit polyclonal) (1:2000) from Santa Cruz Biotechnology (Santa Cruz, CA).

    Techniques: Incubation

    Reciprocal regulation of ERK1/2 and MKP-3. Phosphorylation and activation of ERK1/2 typically occurs through stimulation of the Ras/Raf/MEK1/2 protein kinase cascade. The GTPase Ras activates the protein kinase Raf, which phosphorylates and activates the protein kinases MEK1/2. MEK1/2 is highly specific for phosphorylating and activating ERK1/2. The magnitude and duration of ERK1/2 activity is determined by the balance between the activity of MEK1/2, the kinases that phosphorylate and activate ERK1/2, and the activity of phosphatases, such as MKP-3, which dephosphorylate and inactivate ERK1/2. The levels of MKP-3 protein available to inhibit ERK1/2 are regulated, at least in part, by ERK1/2. ERK1/2 can stimulate an increase in the expression of MKP-3 RNA. This results in an increase in MKP-3 protein, which can then feedback and inhibit ERK1/2 activity. ERK1/2 can also stimulate a decrease in MKP-3 protein stability, however, which can cause a loss of MKP-3 protein. Thus, the levels of MKP-3 protein are determined, at least in part, by the balance between the stimulation of new MKP-3 gene expression and the stability of the MKP-3 protein.

    Journal: Toxicology and applied pharmacology

    Article Title: Reciprocal Regulation of Extracellular Signal Regulated Kinase 1/2 and Mitogen Activated Protein Kinase Phosphatase-3

    doi: 10.1016/j.taap.2008.08.007

    Figure Lengend Snippet: Reciprocal regulation of ERK1/2 and MKP-3. Phosphorylation and activation of ERK1/2 typically occurs through stimulation of the Ras/Raf/MEK1/2 protein kinase cascade. The GTPase Ras activates the protein kinase Raf, which phosphorylates and activates the protein kinases MEK1/2. MEK1/2 is highly specific for phosphorylating and activating ERK1/2. The magnitude and duration of ERK1/2 activity is determined by the balance between the activity of MEK1/2, the kinases that phosphorylate and activate ERK1/2, and the activity of phosphatases, such as MKP-3, which dephosphorylate and inactivate ERK1/2. The levels of MKP-3 protein available to inhibit ERK1/2 are regulated, at least in part, by ERK1/2. ERK1/2 can stimulate an increase in the expression of MKP-3 RNA. This results in an increase in MKP-3 protein, which can then feedback and inhibit ERK1/2 activity. ERK1/2 can also stimulate a decrease in MKP-3 protein stability, however, which can cause a loss of MKP-3 protein. Thus, the levels of MKP-3 protein are determined, at least in part, by the balance between the stimulation of new MKP-3 gene expression and the stability of the MKP-3 protein.

    Article Snippet: After blocking in a TBST/5% milk solution, immunoblots were incubated overnight at 4°C using the following primary antibodies and dilutions: Phospho-p44/42 MAPK (Thr-202/Tyr-204) (E10) (mouse monoclonal) (1:2000), and phospho-MEK1/2 (Ser-217/221) (rabbit polyclonal) (1:2000) from Cell Signaling (Beverly, MA), and ERK2 (C-14) (rabbit polyclonal) (1:2000), MEK1 (12-B) (rabbit polyclonal) (1:500), MKP-3 (C-20) (goat polyclonal) (1:1000) and β-tubulin (H-235) (rabbit polyclonal) (1:2000) from Santa Cruz Biotechnology (Santa Cruz, CA).

    Techniques: Activation Assay, Activity Assay, Expressing

    PathScan MAPK multitarget sandwich ELISA. HOEC monolayers (mixed donors; N = 3) were treated with either TNF-α (10 ng/ml), IL-1β (10 ng/ml), hBD-2 (5 μg/ml), or hBD-3 (5 μg/ml) for 0, 30, or 60 min, respectively. The cell

    Journal: Infection and Immunity

    Article Title: Fusobacterium nucleatum and Human Beta-Defensins Modulate the Release of Antimicrobial Chemokine CCL20/Macrophage Inflammatory Protein 3? ▿ and Human Beta-Defensins Modulate the Release of Antimicrobial Chemokine CCL20/Macrophage Inflammatory Protein 3? ▿ †

    doi: 10.1128/IAI.05586-11

    Figure Lengend Snippet: PathScan MAPK multitarget sandwich ELISA. HOEC monolayers (mixed donors; N = 3) were treated with either TNF-α (10 ng/ml), IL-1β (10 ng/ml), hBD-2 (5 μg/ml), or hBD-3 (5 μg/ml) for 0, 30, or 60 min, respectively. The cell

    Article Snippet: The PathScan multitarget ELISA (targeting phospho-p38, phospho-ERK1/2, phospho-MEK1/2, phospho-SAPK/JNK, and MEK1) ELISA was then performed following the vendor's (Cell signaling, MA) instruction on the clarified supernatant.

    Techniques: Sandwich ELISA

    PathScan MAPK multitarget sandwich ELISA results. HOEC monolayers (mixed donors; N = 3) were treated with either IEF (10 μg/ml) or FnCW (10 μg/ml) for 0, 30, and 60 min. The cell lysates were analyzed for the indicated target, using the

    Journal: Infection and Immunity

    Article Title: Fusobacterium nucleatum and Human Beta-Defensins Modulate the Release of Antimicrobial Chemokine CCL20/Macrophage Inflammatory Protein 3? ▿ and Human Beta-Defensins Modulate the Release of Antimicrobial Chemokine CCL20/Macrophage Inflammatory Protein 3? ▿ †

    doi: 10.1128/IAI.05586-11

    Figure Lengend Snippet: PathScan MAPK multitarget sandwich ELISA results. HOEC monolayers (mixed donors; N = 3) were treated with either IEF (10 μg/ml) or FnCW (10 μg/ml) for 0, 30, and 60 min. The cell lysates were analyzed for the indicated target, using the

    Article Snippet: The PathScan multitarget ELISA (targeting phospho-p38, phospho-ERK1/2, phospho-MEK1/2, phospho-SAPK/JNK, and MEK1) ELISA was then performed following the vendor's (Cell signaling, MA) instruction on the clarified supernatant.

    Techniques: Sandwich ELISA, Electrofocusing

    Inhibitory effect of achillolide A on H 2 O 2 -induced phosphorylation of p44/42 MAPK and MEK1 in astrocytes. Astrocytes were treated with 175 μM of H 2 O 2 for 40 min following preincubation with achillolide A or memantine for 2 h. The levels of phosphorylated and total p44/42 MAPK ( A , B ) and phosphorylated and total MEK1 ( C ) were measured by ELISA. Relative phospho MAPK levels were calculated from the ratio: phospho MAPK levels/total MAPK levels the results are means ± SEM of two experiments ( n = 4). * p

    Journal: Molecules

    Article Title: Achillolide A Protects Astrocytes against Oxidative Stress by Reducing Intracellular Reactive Oxygen Species and Interfering with Cell Signaling

    doi: 10.3390/molecules21030301

    Figure Lengend Snippet: Inhibitory effect of achillolide A on H 2 O 2 -induced phosphorylation of p44/42 MAPK and MEK1 in astrocytes. Astrocytes were treated with 175 μM of H 2 O 2 for 40 min following preincubation with achillolide A or memantine for 2 h. The levels of phosphorylated and total p44/42 MAPK ( A , B ) and phosphorylated and total MEK1 ( C ) were measured by ELISA. Relative phospho MAPK levels were calculated from the ratio: phospho MAPK levels/total MAPK levels the results are means ± SEM of two experiments ( n = 4). * p

    Article Snippet: To measure the amount of total and phosphorylated MEK1 in cell lysates of astrocytes, ELISA was performed according to the manufacturer’s protocol using the PathScan total MEK1 sandwich ELISA kit (Cell Signaling Technology) and the PathScan phosoho-MEK1 (Ser217/221) sandwich ELISA kit (Cell Signaling Technology), respectively.

    Techniques: Enzyme-linked Immunosorbent Assay

    SPI − inhibits LPS-induced NF-κB activation. ApoE–/– mice ( n = 4/diet) were fed CAS- or SPI − -diet for 1 week followed by LPS challenge for 3 h. Nuclear and cytosolic fractions were prepared from aorta and liver lysates (experiment 5). Total- and phospho-NF-κB in nuclear fractions of aorta ( a ) and liver ( b ) were determined by NF-κB Pathscan ELISA kit. Cytosolic I-κB protein levels ( c , d ) in aortic lysates from CAS- or SPI − -fed apoE–/– mice were determined by Western blot. Blots probed with GAPDH were used to normalize I-kB protein levels. Values are mean ± SD, n = 3. Immunofluorescence analysis of NF-κB p65 nuclear translocation in liver CAS- or SPI − -fed mice ( n = 4). Representative pictures from 2 mice per diet group are represented. Liver sections (10 μm) were fixed in PBS/10 % formalin and stained with rabbit anti-NF-κb p65 mAb as described under section “Materials and methods.” Sections were mounted with DAPI to localize nucleus. Number of nucleated (DAPI+) and NF-κB-positive cells (f) were counted in 3 separate sections/mouse. ** P

    Journal: European journal of nutrition

    Article Title: Soy protein inhibits inflammation-induced VCAM-1 and inflammatory cytokine induction by inhibiting the NF-κB and AKT signaling pathway in apolipoprotein E–deficient mice

    doi: 10.1007/s00394-013-0509-7

    Figure Lengend Snippet: SPI − inhibits LPS-induced NF-κB activation. ApoE–/– mice ( n = 4/diet) were fed CAS- or SPI − -diet for 1 week followed by LPS challenge for 3 h. Nuclear and cytosolic fractions were prepared from aorta and liver lysates (experiment 5). Total- and phospho-NF-κB in nuclear fractions of aorta ( a ) and liver ( b ) were determined by NF-κB Pathscan ELISA kit. Cytosolic I-κB protein levels ( c , d ) in aortic lysates from CAS- or SPI − -fed apoE–/– mice were determined by Western blot. Blots probed with GAPDH were used to normalize I-kB protein levels. Values are mean ± SD, n = 3. Immunofluorescence analysis of NF-κB p65 nuclear translocation in liver CAS- or SPI − -fed mice ( n = 4). Representative pictures from 2 mice per diet group are represented. Liver sections (10 μm) were fixed in PBS/10 % formalin and stained with rabbit anti-NF-κb p65 mAb as described under section “Materials and methods.” Sections were mounted with DAPI to localize nucleus. Number of nucleated (DAPI+) and NF-κB-positive cells (f) were counted in 3 separate sections/mouse. ** P

    Article Snippet: Activation of MAP kinase was determined using PathScan multi-target sandwich ELISA kit (Cell signaling) according to the manufacturer's instructions.

    Techniques: Activation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Immunofluorescence, Translocation Assay, Staining

    SPI − inhibits LPS-induced AKT activation. ApoE–/– mice ( n = 4/diet) were fed CAS- or SPI − -diet for 1 week followed by LPS (20 μg/mouse) challenge for 5 h (experiment 5). Total lysates were prepared from liver ( a ) and aorta ( b ), and phosphorylation status of MAP kinase, and AKT were determined using PathScan multi-target sandwich ELISA kit (cell signaling) as described under section “Materials and methods.” Values are mean ± SD, n = 4. * P

    Journal: European journal of nutrition

    Article Title: Soy protein inhibits inflammation-induced VCAM-1 and inflammatory cytokine induction by inhibiting the NF-κB and AKT signaling pathway in apolipoprotein E–deficient mice

    doi: 10.1007/s00394-013-0509-7

    Figure Lengend Snippet: SPI − inhibits LPS-induced AKT activation. ApoE–/– mice ( n = 4/diet) were fed CAS- or SPI − -diet for 1 week followed by LPS (20 μg/mouse) challenge for 5 h (experiment 5). Total lysates were prepared from liver ( a ) and aorta ( b ), and phosphorylation status of MAP kinase, and AKT were determined using PathScan multi-target sandwich ELISA kit (cell signaling) as described under section “Materials and methods.” Values are mean ± SD, n = 4. * P

    Article Snippet: Activation of MAP kinase was determined using PathScan multi-target sandwich ELISA kit (Cell signaling) according to the manufacturer's instructions.

    Techniques: Activation Assay, Mouse Assay, Sandwich ELISA