anti phospho p38 mapk thr180 thr182 rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p38 mapk thr180 thr182 rabbit mab
    a Western blot analysis after SDS-PAGE of <t>phospho-p38</t> levels in 1-day-old wild-type adults with and without cisplatin treatment for indicated times. Worms were exposed to cisplatin on 300 μg/mL cisplatin-containing plates. The blot was <t>probed</t> <t>with</t> <t>anti:phospho-p38</t> antibody to detect activated PMK-1, anti:PMK-1 antibody was used to estimate total levels of PMK-1 protein, tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . b Relative phospho-p38 level quantification from blots presented in Supplementary Fig. . Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. Bars represent mean ± SD. c Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old wild-type adults with (+) and without (−) MitoTempo/cisplatin exposure. The blot was probed with anti:phospho-p38 antibody, tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . d Relative phospho-p38 levels quantification from blots presented in Supplementary Fig. . Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. Bars represent mean ± SD. e , h , i Mean survival ± SD of 1-day-old adults with the indicated genotypes after 24 h cisplatin (300 μg/mL) exposure. Statistical significance determined was by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. All data on cisplatin sensitivity trials is available in Supplementary Data . f , g Relative gene expression of four phase II detoxification genes in 1-day-old adult wild-type animals ( f ) without or with (+CP) 6 h or 24 h cisplatin treatment, g without or with (+CP) 6 h cisplatin treatment. Worms were exposed to cisplatin on 300 μg/mL cisplatin-containing plates. Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate for 6 h exposure timepoint and duplicate for 24 h exposure timepoint. F44B9.5 was used as a normalizing control. Bars represent mean ± SEM. Source data are provided as a Source Data file.
    Anti Phospho P38 Mapk Thr180 Thr182 Rabbit Mab, supplied by Cell Signaling Technology Inc, 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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    Images

    1) Product Images from "Cisplatin toxicity is counteracted by the activation of the p38/ATF-7 signaling pathway in post-mitotic C. elegans"

    Article Title: Cisplatin toxicity is counteracted by the activation of the p38/ATF-7 signaling pathway in post-mitotic C. elegans

    Journal: Nature Communications

    doi: 10.1038/s41467-023-38568-5

    a Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old wild-type adults with and without cisplatin treatment for indicated times. Worms were exposed to cisplatin on 300 μg/mL cisplatin-containing plates. The blot was probed with anti:phospho-p38 antibody to detect activated PMK-1, anti:PMK-1 antibody was used to estimate total levels of PMK-1 protein, tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . b Relative phospho-p38 level quantification from blots presented in Supplementary Fig. . Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. Bars represent mean ± SD. c Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old wild-type adults with (+) and without (−) MitoTempo/cisplatin exposure. The blot was probed with anti:phospho-p38 antibody, tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . d Relative phospho-p38 levels quantification from blots presented in Supplementary Fig. . Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. Bars represent mean ± SD. e , h , i Mean survival ± SD of 1-day-old adults with the indicated genotypes after 24 h cisplatin (300 μg/mL) exposure. Statistical significance determined was by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. All data on cisplatin sensitivity trials is available in Supplementary Data . f , g Relative gene expression of four phase II detoxification genes in 1-day-old adult wild-type animals ( f ) without or with (+CP) 6 h or 24 h cisplatin treatment, g without or with (+CP) 6 h cisplatin treatment. Worms were exposed to cisplatin on 300 μg/mL cisplatin-containing plates. Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate for 6 h exposure timepoint and duplicate for 24 h exposure timepoint. F44B9.5 was used as a normalizing control. Bars represent mean ± SEM. Source data are provided as a Source Data file.
    Figure Legend Snippet: a Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old wild-type adults with and without cisplatin treatment for indicated times. Worms were exposed to cisplatin on 300 μg/mL cisplatin-containing plates. The blot was probed with anti:phospho-p38 antibody to detect activated PMK-1, anti:PMK-1 antibody was used to estimate total levels of PMK-1 protein, tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . b Relative phospho-p38 level quantification from blots presented in Supplementary Fig. . Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. Bars represent mean ± SD. c Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old wild-type adults with (+) and without (−) MitoTempo/cisplatin exposure. The blot was probed with anti:phospho-p38 antibody, tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . d Relative phospho-p38 levels quantification from blots presented in Supplementary Fig. . Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. Bars represent mean ± SD. e , h , i Mean survival ± SD of 1-day-old adults with the indicated genotypes after 24 h cisplatin (300 μg/mL) exposure. Statistical significance determined was by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. All data on cisplatin sensitivity trials is available in Supplementary Data . f , g Relative gene expression of four phase II detoxification genes in 1-day-old adult wild-type animals ( f ) without or with (+CP) 6 h or 24 h cisplatin treatment, g without or with (+CP) 6 h cisplatin treatment. Worms were exposed to cisplatin on 300 μg/mL cisplatin-containing plates. Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate for 6 h exposure timepoint and duplicate for 24 h exposure timepoint. F44B9.5 was used as a normalizing control. Bars represent mean ± SEM. Source data are provided as a Source Data file.

    Techniques Used: Western Blot, SDS Page, Expressing

    a Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old adults with (+) and without (–) 1 mM auxin (AUX) treatment. The blot was probed with anti:phospho-p38 antibody and tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . b , d Mean survival ± SD of 1-day-old adults with the indicated genotypes after 24 h exposure to 300 μg/mL cisplatin-containing plates without (−AUX) or with (+AUX) 1 mM auxin treatment (see the “Methods” section for details). Statistical significance was determined by the b independent two-sided t -test or d one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. c Western blot analysis after SDS-PAGE of SEK-1 levels in 1-day-old adults with (+) and without (−) 1 mM auxin (AUX) treatment. The blot was probed with anti:Flag antibody and tubulin was used as a loading control. Source data are provided as a Source Data file.
    Figure Legend Snippet: a Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old adults with (+) and without (–) 1 mM auxin (AUX) treatment. The blot was probed with anti:phospho-p38 antibody and tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . b , d Mean survival ± SD of 1-day-old adults with the indicated genotypes after 24 h exposure to 300 μg/mL cisplatin-containing plates without (−AUX) or with (+AUX) 1 mM auxin treatment (see the “Methods” section for details). Statistical significance was determined by the b independent two-sided t -test or d one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. c Western blot analysis after SDS-PAGE of SEK-1 levels in 1-day-old adults with (+) and without (−) 1 mM auxin (AUX) treatment. The blot was probed with anti:Flag antibody and tubulin was used as a loading control. Source data are provided as a Source Data file.

    Techniques Used: Western Blot, SDS Page

    anti phospho p38 map kinase thr180 thr182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p38 map kinase thr180 thr182
    A+B: Identification of the Epo-R in skeletal muscle tissue in the non-stimulated state in all 10 subjects with the C20 (A) and M20 (B) antibodies. Positive control is the k-562 cells. M is the molecular marker. C: Activation of the Epo-R by phosphorylation evaluated by western blotting. The samples from the same subject in the stimulated and un-stimulated situation are loaded next to each other D: Phosphorylation of Epo-R, STAT5, <t>p38-MAPK,</t> Akt, p70S6 kinase, Lyn, and IKK, all normalized to the levels of the given protein. The mRNA levels of SOCS3 and IGF-I are normalized to beta-actin mRNA content. Black bars: placebo, white bars: rHuEpo. All results are from biopsies taken 1 h post treatment with either placebo or rHuEpo.
    Anti Phospho P38 Map Kinase Thr180 Thr182, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Evaluation of Functional Erythropoietin Receptor Status in Skeletal Muscle In Vivo: Acute and Prolonged Studies in Healthy Human Subjects"

    Article Title: Evaluation of Functional Erythropoietin Receptor Status in Skeletal Muscle In Vivo: Acute and Prolonged Studies in Healthy Human Subjects

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0031857

    A+B: Identification of the Epo-R in skeletal muscle tissue in the non-stimulated state in all 10 subjects with the C20 (A) and M20 (B) antibodies. Positive control is the k-562 cells. M is the molecular marker. C: Activation of the Epo-R by phosphorylation evaluated by western blotting. The samples from the same subject in the stimulated and un-stimulated situation are loaded next to each other D: Phosphorylation of Epo-R, STAT5, p38-MAPK, Akt, p70S6 kinase, Lyn, and IKK, all normalized to the levels of the given protein. The mRNA levels of SOCS3 and IGF-I are normalized to beta-actin mRNA content. Black bars: placebo, white bars: rHuEpo. All results are from biopsies taken 1 h post treatment with either placebo or rHuEpo.
    Figure Legend Snippet: A+B: Identification of the Epo-R in skeletal muscle tissue in the non-stimulated state in all 10 subjects with the C20 (A) and M20 (B) antibodies. Positive control is the k-562 cells. M is the molecular marker. C: Activation of the Epo-R by phosphorylation evaluated by western blotting. The samples from the same subject in the stimulated and un-stimulated situation are loaded next to each other D: Phosphorylation of Epo-R, STAT5, p38-MAPK, Akt, p70S6 kinase, Lyn, and IKK, all normalized to the levels of the given protein. The mRNA levels of SOCS3 and IGF-I are normalized to beta-actin mRNA content. Black bars: placebo, white bars: rHuEpo. All results are from biopsies taken 1 h post treatment with either placebo or rHuEpo.

    Techniques Used: Positive Control, Marker, Activation Assay, Western Blot

    Phosphorylation of STAT5, p38-MAPK, Akt, p70S6 kinase, Lyn, and IKK, all normalized to the total levels of the given protein. Black bars: placebo, white bars: rHuEpo. The levels of mRNA are measured at 10 h post rHuEpo administration, and are normalized to beta-actin mRNA levels. Level of significance p<0.05, * compared to baseline, # compared to control (same timepoint), § compared to rHuEpo 2 h. The interactions were as follows; pEpo-R p = 0.318, p38-MAPK p = 0.058 (treatment effect p = 0.030), pSTAT5 p = 0.562, p-Akt(ser) p = 0.001, pAkt(thr) p = 0.007, p-IKK p = 0.742 (time effect p = 0.017), p-LYN p = 0.427, p-p70S6kinase p = 0.164 (time effect p = 0.033).
    Figure Legend Snippet: Phosphorylation of STAT5, p38-MAPK, Akt, p70S6 kinase, Lyn, and IKK, all normalized to the total levels of the given protein. Black bars: placebo, white bars: rHuEpo. The levels of mRNA are measured at 10 h post rHuEpo administration, and are normalized to beta-actin mRNA levels. Level of significance p<0.05, * compared to baseline, # compared to control (same timepoint), § compared to rHuEpo 2 h. The interactions were as follows; pEpo-R p = 0.318, p38-MAPK p = 0.058 (treatment effect p = 0.030), pSTAT5 p = 0.562, p-Akt(ser) p = 0.001, pAkt(thr) p = 0.007, p-IKK p = 0.742 (time effect p = 0.017), p-LYN p = 0.427, p-p70S6kinase p = 0.164 (time effect p = 0.033).

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    protein kinase mapk thr180 thr182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc protein kinase mapk thr180 thr182
    Protein Kinase Mapk Thr180 Thr182, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phosphorylated p38 thr180 thr182  (Cell Signaling Technology Inc)


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    anti phospho p38 thr180 thr182 primary antibodies  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p38 thr180 thr182 primary antibodies
    Anti Phospho P38 Thr180 Thr182 Primary Antibodies, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti p p38 thr180 thr182 ab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti p p38 thr180 thr182 ab
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    phospho p38 mapk thr180 thr182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38 mapk thr180 thr182
    Role of the activation of <t>p38</t> <t>MAPK</t> and ERK1/2 in SS2 and PMA induced NETs formation. (A) Neutrophils were infected with SS2 ZY05719 or treated with PMA for 2 h; <t>p38</t> <t>MAPK</t> and ERK1/2 phosphorylation were determined with western blotting using specific antibody against phospho-p38 MAPK (p-p38), p38 <t>MAPK</t> <t>(p38),</t> phospho-ERK1/2 (p-ERK1/2), and ERK1/2. GAPDH was used as an internal control. (B) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and then cells were incubated with SS2 ZY05719 or PMA for 3 h. Results were normalized to neutrophils without pretreatment and activation, and are depicted as the mean ±SEM ( n = 3) of three independent experiments; ** p < 0.01; *** p < 0.001. (C) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and the cells were then incubated with either SS2 ZY05719 or PMA for 3 h. Immunofluorescence was performed using anti-neutrophil elastase (NE) antibody followed by goat anti-rabbit Alexa 568 antibody (red). DNA was stained with DAPI (blue). The results shown are representative of three independent experiments.
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    1) Product Images from "Streptococcus Suis Serotype 2 Stimulates Neutrophil Extracellular Traps Formation via Activation of p38 MAPK and ERK1/2"

    Article Title: Streptococcus Suis Serotype 2 Stimulates Neutrophil Extracellular Traps Formation via Activation of p38 MAPK and ERK1/2

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.02854

    Role of the activation of p38 MAPK and ERK1/2 in SS2 and PMA induced NETs formation. (A) Neutrophils were infected with SS2 ZY05719 or treated with PMA for 2 h; p38 MAPK and ERK1/2 phosphorylation were determined with western blotting using specific antibody against phospho-p38 MAPK (p-p38), p38 MAPK (p38), phospho-ERK1/2 (p-ERK1/2), and ERK1/2. GAPDH was used as an internal control. (B) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and then cells were incubated with SS2 ZY05719 or PMA for 3 h. Results were normalized to neutrophils without pretreatment and activation, and are depicted as the mean ±SEM ( n = 3) of three independent experiments; ** p < 0.01; *** p < 0.001. (C) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and the cells were then incubated with either SS2 ZY05719 or PMA for 3 h. Immunofluorescence was performed using anti-neutrophil elastase (NE) antibody followed by goat anti-rabbit Alexa 568 antibody (red). DNA was stained with DAPI (blue). The results shown are representative of three independent experiments.
    Figure Legend Snippet: Role of the activation of p38 MAPK and ERK1/2 in SS2 and PMA induced NETs formation. (A) Neutrophils were infected with SS2 ZY05719 or treated with PMA for 2 h; p38 MAPK and ERK1/2 phosphorylation were determined with western blotting using specific antibody against phospho-p38 MAPK (p-p38), p38 MAPK (p38), phospho-ERK1/2 (p-ERK1/2), and ERK1/2. GAPDH was used as an internal control. (B) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and then cells were incubated with SS2 ZY05719 or PMA for 3 h. Results were normalized to neutrophils without pretreatment and activation, and are depicted as the mean ±SEM ( n = 3) of three independent experiments; ** p < 0.01; *** p < 0.001. (C) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and the cells were then incubated with either SS2 ZY05719 or PMA for 3 h. Immunofluorescence was performed using anti-neutrophil elastase (NE) antibody followed by goat anti-rabbit Alexa 568 antibody (red). DNA was stained with DAPI (blue). The results shown are representative of three independent experiments.

    Techniques Used: Activation Assay, Infection, Western Blot, Incubation, Immunofluorescence, Staining

    Western blotting analysis of phosphorylation of p38 MAPK and ERK1/2. (A) Neutrophils were pretreated with inhibitors of TLR4 signaling (TAK-242) and NADPH oxidase (DPI) for 30 min, and the cells were then incubated with SS2 ZY05719 and PMA for 2 h. Phosphorylation of p38 MAPK (p-p38) and ERK1/2 (p-ERK1/2) were determined with western blotting. (B) Neutrophils isolated form wild-type mice and TLR knockout mice were incubated with SS2 ZY05719 and Medium.
    Figure Legend Snippet: Western blotting analysis of phosphorylation of p38 MAPK and ERK1/2. (A) Neutrophils were pretreated with inhibitors of TLR4 signaling (TAK-242) and NADPH oxidase (DPI) for 30 min, and the cells were then incubated with SS2 ZY05719 and PMA for 2 h. Phosphorylation of p38 MAPK (p-p38) and ERK1/2 (p-ERK1/2) were determined with western blotting. (B) Neutrophils isolated form wild-type mice and TLR knockout mice were incubated with SS2 ZY05719 and Medium.

    Techniques Used: Western Blot, Incubation, Isolation, Knock-Out

    phospho p38 mapk thr180 thr182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p38 mapk thr180 thr182 rabbit mab
    a Western blot analysis after SDS-PAGE of <t>phospho-p38</t> levels in 1-day-old wild-type adults with and without cisplatin treatment for indicated times. Worms were exposed to cisplatin on 300 μg/mL cisplatin-containing plates. The blot was <t>probed</t> <t>with</t> <t>anti:phospho-p38</t> antibody to detect activated PMK-1, anti:PMK-1 antibody was used to estimate total levels of PMK-1 protein, tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . b Relative phospho-p38 level quantification from blots presented in Supplementary Fig. . Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. Bars represent mean ± SD. c Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old wild-type adults with (+) and without (−) MitoTempo/cisplatin exposure. The blot was probed with anti:phospho-p38 antibody, tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . d Relative phospho-p38 levels quantification from blots presented in Supplementary Fig. . Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. Bars represent mean ± SD. e , h , i Mean survival ± SD of 1-day-old adults with the indicated genotypes after 24 h cisplatin (300 μg/mL) exposure. Statistical significance determined was by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. All data on cisplatin sensitivity trials is available in Supplementary Data . f , g Relative gene expression of four phase II detoxification genes in 1-day-old adult wild-type animals ( f ) without or with (+CP) 6 h or 24 h cisplatin treatment, g without or with (+CP) 6 h cisplatin treatment. Worms were exposed to cisplatin on 300 μg/mL cisplatin-containing plates. Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate for 6 h exposure timepoint and duplicate for 24 h exposure timepoint. F44B9.5 was used as a normalizing control. Bars represent mean ± SEM. Source data are provided as a Source Data file.
    Anti Phospho P38 Mapk Thr180 Thr182 Rabbit Mab, supplied by Cell Signaling Technology Inc, 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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    Cell Signaling Technology Inc anti phospho p38 map kinase thr180 thr182
    A+B: Identification of the Epo-R in skeletal muscle tissue in the non-stimulated state in all 10 subjects with the C20 (A) and M20 (B) antibodies. Positive control is the k-562 cells. M is the molecular marker. C: Activation of the Epo-R by phosphorylation evaluated by western blotting. The samples from the same subject in the stimulated and un-stimulated situation are loaded next to each other D: Phosphorylation of Epo-R, STAT5, <t>p38-MAPK,</t> Akt, p70S6 kinase, Lyn, and IKK, all normalized to the levels of the given protein. The mRNA levels of SOCS3 and IGF-I are normalized to beta-actin mRNA content. Black bars: placebo, white bars: rHuEpo. All results are from biopsies taken 1 h post treatment with either placebo or rHuEpo.
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    A+B: Identification of the Epo-R in skeletal muscle tissue in the non-stimulated state in all 10 subjects with the C20 (A) and M20 (B) antibodies. Positive control is the k-562 cells. M is the molecular marker. C: Activation of the Epo-R by phosphorylation evaluated by western blotting. The samples from the same subject in the stimulated and un-stimulated situation are loaded next to each other D: Phosphorylation of Epo-R, STAT5, <t>p38-MAPK,</t> Akt, p70S6 kinase, Lyn, and IKK, all normalized to the levels of the given protein. The mRNA levels of SOCS3 and IGF-I are normalized to beta-actin mRNA content. Black bars: placebo, white bars: rHuEpo. All results are from biopsies taken 1 h post treatment with either placebo or rHuEpo.
    Protein Kinase Mapk Thr180 Thr182, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    A+B: Identification of the Epo-R in skeletal muscle tissue in the non-stimulated state in all 10 subjects with the C20 (A) and M20 (B) antibodies. Positive control is the k-562 cells. M is the molecular marker. C: Activation of the Epo-R by phosphorylation evaluated by western blotting. The samples from the same subject in the stimulated and un-stimulated situation are loaded next to each other D: Phosphorylation of Epo-R, STAT5, <t>p38-MAPK,</t> Akt, p70S6 kinase, Lyn, and IKK, all normalized to the levels of the given protein. The mRNA levels of SOCS3 and IGF-I are normalized to beta-actin mRNA content. Black bars: placebo, white bars: rHuEpo. All results are from biopsies taken 1 h post treatment with either placebo or rHuEpo.
    Phosphorylated P38 Thr180 Thr182, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    A+B: Identification of the Epo-R in skeletal muscle tissue in the non-stimulated state in all 10 subjects with the C20 (A) and M20 (B) antibodies. Positive control is the k-562 cells. M is the molecular marker. C: Activation of the Epo-R by phosphorylation evaluated by western blotting. The samples from the same subject in the stimulated and un-stimulated situation are loaded next to each other D: Phosphorylation of Epo-R, STAT5, <t>p38-MAPK,</t> Akt, p70S6 kinase, Lyn, and IKK, all normalized to the levels of the given protein. The mRNA levels of SOCS3 and IGF-I are normalized to beta-actin mRNA content. Black bars: placebo, white bars: rHuEpo. All results are from biopsies taken 1 h post treatment with either placebo or rHuEpo.
    Anti Phospho P38 Thr180 Thr182 Primary Antibodies, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    A+B: Identification of the Epo-R in skeletal muscle tissue in the non-stimulated state in all 10 subjects with the C20 (A) and M20 (B) antibodies. Positive control is the k-562 cells. M is the molecular marker. C: Activation of the Epo-R by phosphorylation evaluated by western blotting. The samples from the same subject in the stimulated and un-stimulated situation are loaded next to each other D: Phosphorylation of Epo-R, STAT5, <t>p38-MAPK,</t> Akt, p70S6 kinase, Lyn, and IKK, all normalized to the levels of the given protein. The mRNA levels of SOCS3 and IGF-I are normalized to beta-actin mRNA content. Black bars: placebo, white bars: rHuEpo. All results are from biopsies taken 1 h post treatment with either placebo or rHuEpo.
    Anti P P38 Thr180 Thr182 Ab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Role of the activation of <t>p38</t> <t>MAPK</t> and ERK1/2 in SS2 and PMA induced NETs formation. (A) Neutrophils were infected with SS2 ZY05719 or treated with PMA for 2 h; <t>p38</t> <t>MAPK</t> and ERK1/2 phosphorylation were determined with western blotting using specific antibody against phospho-p38 MAPK (p-p38), p38 <t>MAPK</t> <t>(p38),</t> phospho-ERK1/2 (p-ERK1/2), and ERK1/2. GAPDH was used as an internal control. (B) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and then cells were incubated with SS2 ZY05719 or PMA for 3 h. Results were normalized to neutrophils without pretreatment and activation, and are depicted as the mean ±SEM ( n = 3) of three independent experiments; ** p < 0.01; *** p < 0.001. (C) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and the cells were then incubated with either SS2 ZY05719 or PMA for 3 h. Immunofluorescence was performed using anti-neutrophil elastase (NE) antibody followed by goat anti-rabbit Alexa 568 antibody (red). DNA was stained with DAPI (blue). The results shown are representative of three independent experiments.
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    Role of the activation of <t>p38</t> <t>MAPK</t> and ERK1/2 in SS2 and PMA induced NETs formation. (A) Neutrophils were infected with SS2 ZY05719 or treated with PMA for 2 h; <t>p38</t> <t>MAPK</t> and ERK1/2 phosphorylation were determined with western blotting using specific antibody against phospho-p38 MAPK (p-p38), p38 <t>MAPK</t> <t>(p38),</t> phospho-ERK1/2 (p-ERK1/2), and ERK1/2. GAPDH was used as an internal control. (B) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and then cells were incubated with SS2 ZY05719 or PMA for 3 h. Results were normalized to neutrophils without pretreatment and activation, and are depicted as the mean ±SEM ( n = 3) of three independent experiments; ** p < 0.01; *** p < 0.001. (C) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and the cells were then incubated with either SS2 ZY05719 or PMA for 3 h. Immunofluorescence was performed using anti-neutrophil elastase (NE) antibody followed by goat anti-rabbit Alexa 568 antibody (red). DNA was stained with DAPI (blue). The results shown are representative of three independent experiments.
    Anti Phospho P38 Thr180 Thr182, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Role of the activation of <t>p38</t> <t>MAPK</t> and ERK1/2 in SS2 and PMA induced NETs formation. (A) Neutrophils were infected with SS2 ZY05719 or treated with PMA for 2 h; <t>p38</t> <t>MAPK</t> and ERK1/2 phosphorylation were determined with western blotting using specific antibody against phospho-p38 MAPK (p-p38), p38 <t>MAPK</t> <t>(p38),</t> phospho-ERK1/2 (p-ERK1/2), and ERK1/2. GAPDH was used as an internal control. (B) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and then cells were incubated with SS2 ZY05719 or PMA for 3 h. Results were normalized to neutrophils without pretreatment and activation, and are depicted as the mean ±SEM ( n = 3) of three independent experiments; ** p < 0.01; *** p < 0.001. (C) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and the cells were then incubated with either SS2 ZY05719 or PMA for 3 h. Immunofluorescence was performed using anti-neutrophil elastase (NE) antibody followed by goat anti-rabbit Alexa 568 antibody (red). DNA was stained with DAPI (blue). The results shown are representative of three independent experiments.
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    Image Search Results


    a Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old wild-type adults with and without cisplatin treatment for indicated times. Worms were exposed to cisplatin on 300 μg/mL cisplatin-containing plates. The blot was probed with anti:phospho-p38 antibody to detect activated PMK-1, anti:PMK-1 antibody was used to estimate total levels of PMK-1 protein, tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . b Relative phospho-p38 level quantification from blots presented in Supplementary Fig. . Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. Bars represent mean ± SD. c Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old wild-type adults with (+) and without (−) MitoTempo/cisplatin exposure. The blot was probed with anti:phospho-p38 antibody, tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . d Relative phospho-p38 levels quantification from blots presented in Supplementary Fig. . Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. Bars represent mean ± SD. e , h , i Mean survival ± SD of 1-day-old adults with the indicated genotypes after 24 h cisplatin (300 μg/mL) exposure. Statistical significance determined was by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. All data on cisplatin sensitivity trials is available in Supplementary Data . f , g Relative gene expression of four phase II detoxification genes in 1-day-old adult wild-type animals ( f ) without or with (+CP) 6 h or 24 h cisplatin treatment, g without or with (+CP) 6 h cisplatin treatment. Worms were exposed to cisplatin on 300 μg/mL cisplatin-containing plates. Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate for 6 h exposure timepoint and duplicate for 24 h exposure timepoint. F44B9.5 was used as a normalizing control. Bars represent mean ± SEM. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Cisplatin toxicity is counteracted by the activation of the p38/ATF-7 signaling pathway in post-mitotic C. elegans

    doi: 10.1038/s41467-023-38568-5

    Figure Lengend Snippet: a Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old wild-type adults with and without cisplatin treatment for indicated times. Worms were exposed to cisplatin on 300 μg/mL cisplatin-containing plates. The blot was probed with anti:phospho-p38 antibody to detect activated PMK-1, anti:PMK-1 antibody was used to estimate total levels of PMK-1 protein, tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . b Relative phospho-p38 level quantification from blots presented in Supplementary Fig. . Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. Bars represent mean ± SD. c Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old wild-type adults with (+) and without (−) MitoTempo/cisplatin exposure. The blot was probed with anti:phospho-p38 antibody, tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . d Relative phospho-p38 levels quantification from blots presented in Supplementary Fig. . Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. Bars represent mean ± SD. e , h , i Mean survival ± SD of 1-day-old adults with the indicated genotypes after 24 h cisplatin (300 μg/mL) exposure. Statistical significance determined was by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. All data on cisplatin sensitivity trials is available in Supplementary Data . f , g Relative gene expression of four phase II detoxification genes in 1-day-old adult wild-type animals ( f ) without or with (+CP) 6 h or 24 h cisplatin treatment, g without or with (+CP) 6 h cisplatin treatment. Worms were exposed to cisplatin on 300 μg/mL cisplatin-containing plates. Statistical significance was determined by one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate for 6 h exposure timepoint and duplicate for 24 h exposure timepoint. F44B9.5 was used as a normalizing control. Bars represent mean ± SEM. Source data are provided as a Source Data file.

    Article Snippet: Primary antibodies were: anti:phospho-p38 MAPK (Thr180/Thr182) Rabbit mAb (D3F9, Cell Signaling) 1:1000, anti:total PMK-1 MAPK Rabbit pAb 1:1000 from the Pukkila-Worley lab , anti:tubulin (T5168, Sigma) 1:5000, anti:Flag (M2, Sigma) 1:1000.

    Techniques: Western Blot, SDS Page, Expressing

    a Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old adults with (+) and without (–) 1 mM auxin (AUX) treatment. The blot was probed with anti:phospho-p38 antibody and tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . b , d Mean survival ± SD of 1-day-old adults with the indicated genotypes after 24 h exposure to 300 μg/mL cisplatin-containing plates without (−AUX) or with (+AUX) 1 mM auxin treatment (see the “Methods” section for details). Statistical significance was determined by the b independent two-sided t -test or d one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. c Western blot analysis after SDS-PAGE of SEK-1 levels in 1-day-old adults with (+) and without (−) 1 mM auxin (AUX) treatment. The blot was probed with anti:Flag antibody and tubulin was used as a loading control. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Cisplatin toxicity is counteracted by the activation of the p38/ATF-7 signaling pathway in post-mitotic C. elegans

    doi: 10.1038/s41467-023-38568-5

    Figure Lengend Snippet: a Western blot analysis after SDS-PAGE of phospho-p38 levels in 1-day-old adults with (+) and without (–) 1 mM auxin (AUX) treatment. The blot was probed with anti:phospho-p38 antibody and tubulin was used as a loading control. Full uncropped images are available in Supplementary Fig. . b , d Mean survival ± SD of 1-day-old adults with the indicated genotypes after 24 h exposure to 300 μg/mL cisplatin-containing plates without (−AUX) or with (+AUX) 1 mM auxin treatment (see the “Methods” section for details). Statistical significance was determined by the b independent two-sided t -test or d one-way ANOVA followed by Bonferroni post hoc correction. The experiment was performed in triplicate. c Western blot analysis after SDS-PAGE of SEK-1 levels in 1-day-old adults with (+) and without (−) 1 mM auxin (AUX) treatment. The blot was probed with anti:Flag antibody and tubulin was used as a loading control. Source data are provided as a Source Data file.

    Article Snippet: Primary antibodies were: anti:phospho-p38 MAPK (Thr180/Thr182) Rabbit mAb (D3F9, Cell Signaling) 1:1000, anti:total PMK-1 MAPK Rabbit pAb 1:1000 from the Pukkila-Worley lab , anti:tubulin (T5168, Sigma) 1:5000, anti:Flag (M2, Sigma) 1:1000.

    Techniques: Western Blot, SDS Page

    A+B: Identification of the Epo-R in skeletal muscle tissue in the non-stimulated state in all 10 subjects with the C20 (A) and M20 (B) antibodies. Positive control is the k-562 cells. M is the molecular marker. C: Activation of the Epo-R by phosphorylation evaluated by western blotting. The samples from the same subject in the stimulated and un-stimulated situation are loaded next to each other D: Phosphorylation of Epo-R, STAT5, p38-MAPK, Akt, p70S6 kinase, Lyn, and IKK, all normalized to the levels of the given protein. The mRNA levels of SOCS3 and IGF-I are normalized to beta-actin mRNA content. Black bars: placebo, white bars: rHuEpo. All results are from biopsies taken 1 h post treatment with either placebo or rHuEpo.

    Journal: PLoS ONE

    Article Title: Evaluation of Functional Erythropoietin Receptor Status in Skeletal Muscle In Vivo: Acute and Prolonged Studies in Healthy Human Subjects

    doi: 10.1371/journal.pone.0031857

    Figure Lengend Snippet: A+B: Identification of the Epo-R in skeletal muscle tissue in the non-stimulated state in all 10 subjects with the C20 (A) and M20 (B) antibodies. Positive control is the k-562 cells. M is the molecular marker. C: Activation of the Epo-R by phosphorylation evaluated by western blotting. The samples from the same subject in the stimulated and un-stimulated situation are loaded next to each other D: Phosphorylation of Epo-R, STAT5, p38-MAPK, Akt, p70S6 kinase, Lyn, and IKK, all normalized to the levels of the given protein. The mRNA levels of SOCS3 and IGF-I are normalized to beta-actin mRNA content. Black bars: placebo, white bars: rHuEpo. All results are from biopsies taken 1 h post treatment with either placebo or rHuEpo.

    Article Snippet: The protein fraction was analysed for phosphorylation of Epo-R, Lyn, STAT5, Akt, p70S6-kinase, MAPK and for total Epo-R. Primary antibodies were as follows: anti-phospho-Epo-R(Tyr456) (Santa Cruz, #sc20236), anti-Lyn (Cell signalling, #2732), anti-phospho-Lyn(Tyr507) (Cell signalling, #2731), anti-STAT5 (Cell signalling, #9310), anti-phospho-STAT5(Tyr694) (Cell signalling, #9351), anti-Akt/PKB (Cell signalling, #9272), anti-phospho-Akt/PKB(Ser473) (Cell signalling, #9271), anti-phospho-Akt/PKB(Thr308) (Cell signalling, #9275), anti-p70S6 kinase (Cell signalling, #9202), anti-phospho-p70S6 kinase(Thr389) (Cell signalling, #9205), anti-p38-MAP kinase (Cell signalling, #9212), anti-phospho-p38-MAP kinase (Thr180/Thr182) (Cell signalling, #9211), anti-IKKα (Cell signalling, #2682), anti-phospho-IKKα/β(Ser176/180) (Cell signalling, #2697), anti-Epo-R (M20) (Santa Cruz, #sc697), anti-Epo-R (C20) (Santa Cruz, #sc695), and anti-β-actin (Abcam, #ab8227).

    Techniques: Positive Control, Marker, Activation Assay, Western Blot

    Phosphorylation of STAT5, p38-MAPK, Akt, p70S6 kinase, Lyn, and IKK, all normalized to the total levels of the given protein. Black bars: placebo, white bars: rHuEpo. The levels of mRNA are measured at 10 h post rHuEpo administration, and are normalized to beta-actin mRNA levels. Level of significance p<0.05, * compared to baseline, # compared to control (same timepoint), § compared to rHuEpo 2 h. The interactions were as follows; pEpo-R p = 0.318, p38-MAPK p = 0.058 (treatment effect p = 0.030), pSTAT5 p = 0.562, p-Akt(ser) p = 0.001, pAkt(thr) p = 0.007, p-IKK p = 0.742 (time effect p = 0.017), p-LYN p = 0.427, p-p70S6kinase p = 0.164 (time effect p = 0.033).

    Journal: PLoS ONE

    Article Title: Evaluation of Functional Erythropoietin Receptor Status in Skeletal Muscle In Vivo: Acute and Prolonged Studies in Healthy Human Subjects

    doi: 10.1371/journal.pone.0031857

    Figure Lengend Snippet: Phosphorylation of STAT5, p38-MAPK, Akt, p70S6 kinase, Lyn, and IKK, all normalized to the total levels of the given protein. Black bars: placebo, white bars: rHuEpo. The levels of mRNA are measured at 10 h post rHuEpo administration, and are normalized to beta-actin mRNA levels. Level of significance p<0.05, * compared to baseline, # compared to control (same timepoint), § compared to rHuEpo 2 h. The interactions were as follows; pEpo-R p = 0.318, p38-MAPK p = 0.058 (treatment effect p = 0.030), pSTAT5 p = 0.562, p-Akt(ser) p = 0.001, pAkt(thr) p = 0.007, p-IKK p = 0.742 (time effect p = 0.017), p-LYN p = 0.427, p-p70S6kinase p = 0.164 (time effect p = 0.033).

    Article Snippet: The protein fraction was analysed for phosphorylation of Epo-R, Lyn, STAT5, Akt, p70S6-kinase, MAPK and for total Epo-R. Primary antibodies were as follows: anti-phospho-Epo-R(Tyr456) (Santa Cruz, #sc20236), anti-Lyn (Cell signalling, #2732), anti-phospho-Lyn(Tyr507) (Cell signalling, #2731), anti-STAT5 (Cell signalling, #9310), anti-phospho-STAT5(Tyr694) (Cell signalling, #9351), anti-Akt/PKB (Cell signalling, #9272), anti-phospho-Akt/PKB(Ser473) (Cell signalling, #9271), anti-phospho-Akt/PKB(Thr308) (Cell signalling, #9275), anti-p70S6 kinase (Cell signalling, #9202), anti-phospho-p70S6 kinase(Thr389) (Cell signalling, #9205), anti-p38-MAP kinase (Cell signalling, #9212), anti-phospho-p38-MAP kinase (Thr180/Thr182) (Cell signalling, #9211), anti-IKKα (Cell signalling, #2682), anti-phospho-IKKα/β(Ser176/180) (Cell signalling, #2697), anti-Epo-R (M20) (Santa Cruz, #sc697), anti-Epo-R (C20) (Santa Cruz, #sc695), and anti-β-actin (Abcam, #ab8227).

    Techniques:

    Role of the activation of p38 MAPK and ERK1/2 in SS2 and PMA induced NETs formation. (A) Neutrophils were infected with SS2 ZY05719 or treated with PMA for 2 h; p38 MAPK and ERK1/2 phosphorylation were determined with western blotting using specific antibody against phospho-p38 MAPK (p-p38), p38 MAPK (p38), phospho-ERK1/2 (p-ERK1/2), and ERK1/2. GAPDH was used as an internal control. (B) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and then cells were incubated with SS2 ZY05719 or PMA for 3 h. Results were normalized to neutrophils without pretreatment and activation, and are depicted as the mean ±SEM ( n = 3) of three independent experiments; ** p < 0.01; *** p < 0.001. (C) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and the cells were then incubated with either SS2 ZY05719 or PMA for 3 h. Immunofluorescence was performed using anti-neutrophil elastase (NE) antibody followed by goat anti-rabbit Alexa 568 antibody (red). DNA was stained with DAPI (blue). The results shown are representative of three independent experiments.

    Journal: Frontiers in Immunology

    Article Title: Streptococcus Suis Serotype 2 Stimulates Neutrophil Extracellular Traps Formation via Activation of p38 MAPK and ERK1/2

    doi: 10.3389/fimmu.2018.02854

    Figure Lengend Snippet: Role of the activation of p38 MAPK and ERK1/2 in SS2 and PMA induced NETs formation. (A) Neutrophils were infected with SS2 ZY05719 or treated with PMA for 2 h; p38 MAPK and ERK1/2 phosphorylation were determined with western blotting using specific antibody against phospho-p38 MAPK (p-p38), p38 MAPK (p38), phospho-ERK1/2 (p-ERK1/2), and ERK1/2. GAPDH was used as an internal control. (B) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and then cells were incubated with SS2 ZY05719 or PMA for 3 h. Results were normalized to neutrophils without pretreatment and activation, and are depicted as the mean ±SEM ( n = 3) of three independent experiments; ** p < 0.01; *** p < 0.001. (C) Neutrophils were pretreated with inhibitor of p38 MAPK (SB203580) and ERK1/2 (U0126), and the cells were then incubated with either SS2 ZY05719 or PMA for 3 h. Immunofluorescence was performed using anti-neutrophil elastase (NE) antibody followed by goat anti-rabbit Alexa 568 antibody (red). DNA was stained with DAPI (blue). The results shown are representative of three independent experiments.

    Article Snippet: Thereafter, the membranes were incubated with Phospho-p38 MAPK (Thr180/Thr182) (D3F9) XP ® Rabbit mAb, p38 MAPK (D13E1) XP® Rabbit mAb, Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E)XP ® Rabbit mAb, p44/42 MAPK (Erk1/2)(137F5) Rabbit mAb, Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb (1:1000, Cell Signaling Technology, Danvers, MA, USA), or anti-GAPDH Rabbit pAb (1:5000, CMCTAG, Milwaukee, WI, USA) at 4°C overnight.

    Techniques: Activation Assay, Infection, Western Blot, Incubation, Immunofluorescence, Staining

    Western blotting analysis of phosphorylation of p38 MAPK and ERK1/2. (A) Neutrophils were pretreated with inhibitors of TLR4 signaling (TAK-242) and NADPH oxidase (DPI) for 30 min, and the cells were then incubated with SS2 ZY05719 and PMA for 2 h. Phosphorylation of p38 MAPK (p-p38) and ERK1/2 (p-ERK1/2) were determined with western blotting. (B) Neutrophils isolated form wild-type mice and TLR knockout mice were incubated with SS2 ZY05719 and Medium.

    Journal: Frontiers in Immunology

    Article Title: Streptococcus Suis Serotype 2 Stimulates Neutrophil Extracellular Traps Formation via Activation of p38 MAPK and ERK1/2

    doi: 10.3389/fimmu.2018.02854

    Figure Lengend Snippet: Western blotting analysis of phosphorylation of p38 MAPK and ERK1/2. (A) Neutrophils were pretreated with inhibitors of TLR4 signaling (TAK-242) and NADPH oxidase (DPI) for 30 min, and the cells were then incubated with SS2 ZY05719 and PMA for 2 h. Phosphorylation of p38 MAPK (p-p38) and ERK1/2 (p-ERK1/2) were determined with western blotting. (B) Neutrophils isolated form wild-type mice and TLR knockout mice were incubated with SS2 ZY05719 and Medium.

    Article Snippet: Thereafter, the membranes were incubated with Phospho-p38 MAPK (Thr180/Thr182) (D3F9) XP ® Rabbit mAb, p38 MAPK (D13E1) XP® Rabbit mAb, Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E)XP ® Rabbit mAb, p44/42 MAPK (Erk1/2)(137F5) Rabbit mAb, Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb (1:1000, Cell Signaling Technology, Danvers, MA, USA), or anti-GAPDH Rabbit pAb (1:5000, CMCTAG, Milwaukee, WI, USA) at 4°C overnight.

    Techniques: Western Blot, Incubation, Isolation, Knock-Out