STEP Antibody (23E5) - BSA Free

Western Blot: STEP Antibody (23E5) - BSA Free [NB300-202] -

Glutamate induces sustained p38 MAPK phosphorylation in STEP-deficient neurons. Neuronal cultures from (A and C) WT and (B and D) and STEP KO mice were treated with 50 μM glutamate (Glu) or 50 μM NMDA for the specified times. A–D, equal amounts of protein from each sample were analyzed by immunoblot analysis using anti-phospho-p38 (top), anti-p38 (middle), and anti-STEP (bottom) antibodies. E, neuronal cultures from STEP KO mice were treated with glutamate (50 μM) for 20 min in the absence or presence of MK801 (5 μM). Protein extracts were analyzed by immunoblotting with anti-phospho-p38 (top) and anti-p38 (bottom) antibodies. Corresponding bar diagrams represent quantitative analysis of p38 MAPK phosphorylation as the mean +/- SD (n = 3). ∗p < 0.01 and ∗∗p < 0.001 compared with the untreated control. Statistical analysis has been performed using ANOVA with Tukey’s post hoc test. #p < 0.001 and ##p < 0.0001 from 5 min glutamate treatment. NMDA, N-methyl-D-aspartic acid; p38 MAPK, p38 mitogen-activated protein kinase; STEP, striatal-enriched phosphatase. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34246631), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: STEP Antibody (23E5) - BSA Free [NB300-202] -

Role of the NMDAR and p38 MAPK in glutamate-induced increase in the COX-2 protein level in STEP-deficient neurons. Neuronal cultures from (A) WT and (B) STEP KO mice were treated with 50 μM glutamate (Glu) for 20 min and then maintained in their original medium for the specified times (post-Glu time). C and D, neuronal cultures from STEP KO mice were treated with 50 μM glutamate (Glu) for 20 min followed by recovery (post-Glu time) in the absence and presence of (C) MK801 (5 μM) or (D) SB 203580 (5 μM). Equal amounts of protein from each sample were analyzed by immunoblotting using anti-COX-2 (top) and anti-beta -tubulin (bottom) antibodies. Corresponding bar diagrams represents quantitative analysis of COX-2 protein level as the mean +/- SD (n = 3–4). Statistical analysis has been performed using ANOVA with Tukey’s post hoc test. ∗p < 0.001 and ∗∗p < 0.0001 compared with untreated control and #p < 0.01 and ##p < 0.001 from 2 h postglutamate time. COX-2, cyclooxygenase-2; p38 MAPK, p38 mitogen-activated protein kinase; STEP, striatal-enriched phosphatase. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34246631), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: STEP Antibody (23E5) - BSA Free [NB300-202] -

Glutamate induces sustained p38 MAPK phosphorylation in STEP-deficient neurons. Neuronal cultures from (A and C) WT and (B and D) and STEP KO mice were treated with 50 μM glutamate (Glu) or 50 μM NMDA for the specified times. A–D, equal amounts of protein from each sample were analyzed by immunoblot analysis using anti-phospho-p38 (top), anti-p38 (middle), and anti-STEP (bottom) antibodies. E, neuronal cultures from STEP KO mice were treated with glutamate (50 μM) for 20 min in the absence or presence of MK801 (5 μM). Protein extracts were analyzed by immunoblotting with anti-phospho-p38 (top) and anti-p38 (bottom) antibodies. Corresponding bar diagrams represent quantitative analysis of p38 MAPK phosphorylation as the mean +/- SD (n = 3). ∗p < 0.01 and ∗∗p < 0.001 compared with the untreated control. Statistical analysis has been performed using ANOVA with Tukey’s post hoc test. #p < 0.001 and ##p < 0.0001 from 5 min glutamate treatment. NMDA, N-methyl-D-aspartic acid; p38 MAPK, p38 mitogen-activated protein kinase; STEP, striatal-enriched phosphatase. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34246631), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: STEP Antibody (23E5) - BSA Free [NB300-202] -

Glutamate induces sustained p38 MAPK phosphorylation in STEP-deficient neurons. Neuronal cultures from (A and C) WT and (B and D) and STEP KO mice were treated with 50 μM glutamate (Glu) or 50 μM NMDA for the specified times. A–D, equal amounts of protein from each sample were analyzed by immunoblot analysis using anti-phospho-p38 (top), anti-p38 (middle), and anti-STEP (bottom) antibodies. E, neuronal cultures from STEP KO mice were treated with glutamate (50 μM) for 20 min in the absence or presence of MK801 (5 μM). Protein extracts were analyzed by immunoblotting with anti-phospho-p38 (top) and anti-p38 (bottom) antibodies. Corresponding bar diagrams represent quantitative analysis of p38 MAPK phosphorylation as the mean +/- SD (n = 3). ∗p < 0.01 and ∗∗p < 0.001 compared with the untreated control. Statistical analysis has been performed using ANOVA with Tukey’s post hoc test. #p < 0.001 and ##p < 0.0001 from 5 min glutamate treatment. NMDA, N-methyl-D-aspartic acid; p38 MAPK, p38 mitogen-activated protein kinase; STEP, striatal-enriched phosphatase. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34246631), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: STEP Antibody (23E5) - BSA Free [NB300-202] -

Role of NF-kappa B in glutamate-induced increase in the COX-2 protein level in STEP-deficient neurons.A and B, neuronal cultures from (A) WT and (B) STEP KO mice were treated with 50 μM glutamate (Glu) for 20 min and then maintained in their original medium for the specified times (post-Glu time). C–F, neuronal cultures from STEP KO mice were treated with 50 μM glutamate (Glu) for 20 min followed by recovery (post-Glu time) in the absence and presence of (C) MK801 (5 μM), (D) SB 203580 (5 μM), or (E and F) Bengamide B (500 nM). Equal amounts of protein from each sample were analyzed by immunoblotting using (A–D) anti-I kappaB alpha (top) and anti-beta -tubulin (bottom) antibodies, or (E) anti-COX-2 (top) and anti-beta -tubulin (bottom) antibodies. Corresponding bar diagrams represents quantitative analysis of (A–D) I kappaB alpha or (E) COX-2 protein levels as the mean +/- SD. F, equal amounts of culture media from each sample were analyzed for PGE2 level using ELISA. Statistical analysis has been performed using ANOVA with Tukey’s post hoc test. Values are expressed as the mean +/- SD (n = 3–4). ∗p < 0.01 and ∗∗p < 0.001 compared with the untreated control and #p < 0.01 from 2 h postglutamate time. COX-2, cyclooxygenase-2; I kappaB, inhibitor of nuclear factor-kappa B; STEP, striatal-enriched phosphatase. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34246631), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: STEP Antibody (23E5) - BSA Free [NB300-202] -

Sustained p38 MAPK phosphorylation in STEP-deficient neurons in the postglutamate recovery phase. Neuronal cultures from (A) WT and (B) and STEP KO mice were treated with 50 μM glutamate (Glu) for 20 min and then maintained in their original medium for the specified times (post-Glu time). Protein extracts were analyzed by immunoblotting with anti-phospho-p38 (top), anti-p38 (middle), and anti-STEP (bottom) antibodies. Corresponding bar diagrams represent quantitative analysis of p38 MAPK phosphorylation as the mean +/- SD (n = 3). Statistical analysis has been performed using ANOVA with Tukey’s post hoc test. ∗p < 0.01 and ∗∗p < 0.001 compared with the untreated control. #p < 0.01 from 20 min glutamate treatment. p38 MAPK, p38 mitogen-activated protein kinase; STEP, striatal-enriched phosphatase. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34246631), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: STEP Antibody (23E5) - BSA Free [NB300-202] -

A STEP mimetic attenuates glutamate-induced increase in p38 MAPK phosphorylation, COX-2 expression, and PGE2release in STEP-deficient neurons.A, schematic representation of TAT-STEP-Myc peptide generated from STEP61. The diagram of STEP61 shows the positions of the phosphatase domain, transmembrane domain (TM), kinase-interaction motif (KIM), kinase specificity sequence (KIS), and the phosphorylation sites in the KIM and KIS domains. The diagram of the TAT-STEP-Myc peptide (STEP mimetic), derived from STEP61, shows the positions of the TAT domain at the N terminus, myc-tag at the C terminus, the serine residue in the KIM domain that was mutated to alanine to allow the peptide to bind to its substrates, and the threonine and serine residues in the KIS domain, which were mutated to glutamic acid to render the peptide resistant to degradation. B–D, neuronal cultures from STEP KO mice were treated with 50 μM glutamate (Glu) for 20 min in the absence and presence of TAT-STEP-myc peptide and then maintained in their original medium for 2 h (post-Glu time). B and C, equal amounts of protein from each sample were analyzed by immunoblotting using anti-phospho-p38 (top) and anti-p38 (bottom) antibodies. Corresponding bar diagrams represent quantitative analysis of p38 MAPK phosphorylation as the mean +/- SD (n = 3–4). D, equal amounts of culture media from each sample were analyzed for the PGE2 level using ELISA. Statistical analysis has been performed using ANOVA with Tukey’s post hoc test. Values are expressed as the mean +/- SD (n = 4). ∗p < 0.01 and ∗∗p < 0.001 compared with the untreated control. #p < 0.05 and ##p < 0.01 from 2 h post-glutamate time. COX-2, cyclooxygenase-2; p38 MAPK, p38 mitogen-activated protein kinase; STEP, striatal-enriched phosphatase. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34246631), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: STEP Antibody (23E5) - BSA Free [NB300-202] -

A STEP mimetic attenuates glutamate-induced increase in p38 MAPK phosphorylation, COX-2 expression, and PGE2release in STEP-deficient neurons.A, schematic representation of TAT-STEP-Myc peptide generated from STEP61. The diagram of STEP61 shows the positions of the phosphatase domain, transmembrane domain (TM), kinase-interaction motif (KIM), kinase specificity sequence (KIS), and the phosphorylation sites in the KIM and KIS domains. The diagram of the TAT-STEP-Myc peptide (STEP mimetic), derived from STEP61, shows the positions of the TAT domain at the N terminus, myc-tag at the C terminus, the serine residue in the KIM domain that was mutated to alanine to allow the peptide to bind to its substrates, and the threonine and serine residues in the KIS domain, which were mutated to glutamic acid to render the peptide resistant to degradation. B–D, neuronal cultures from STEP KO mice were treated with 50 μM glutamate (Glu) for 20 min in the absence and presence of TAT-STEP-myc peptide and then maintained in their original medium for 2 h (post-Glu time). B and C, equal amounts of protein from each sample were analyzed by immunoblotting using anti-phospho-p38 (top) and anti-p38 (bottom) antibodies. Corresponding bar diagrams represent quantitative analysis of p38 MAPK phosphorylation as the mean +/- SD (n = 3–4). D, equal amounts of culture media from each sample were analyzed for the PGE2 level using ELISA. Statistical analysis has been performed using ANOVA with Tukey’s post hoc test. Values are expressed as the mean +/- SD (n = 4). ∗p < 0.01 and ∗∗p < 0.001 compared with the untreated control. #p < 0.05 and ##p < 0.01 from 2 h post-glutamate time. COX-2, cyclooxygenase-2; p38 MAPK, p38 mitogen-activated protein kinase; STEP, striatal-enriched phosphatase. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34246631), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: STEP Antibody (23E5) - BSA Free [NB300-202] -

Role of the NMDAR and p38 MAPK in glutamate-induced increase in the COX-2 protein level in STEP-deficient neurons. Neuronal cultures from (A) WT and (B) STEP KO mice were treated with 50 μM glutamate (Glu) for 20 min and then maintained in their original medium for the specified times (post-Glu time). C and D, neuronal cultures from STEP KO mice were treated with 50 μM glutamate (Glu) for 20 min followed by recovery (post-Glu time) in the absence and presence of (C) MK801 (5 μM) or (D) SB 203580 (5 μM). Equal amounts of protein from each sample were analyzed by immunoblotting using anti-COX-2 (top) and anti-beta -tubulin (bottom) antibodies. Corresponding bar diagrams represents quantitative analysis of COX-2 protein level as the mean +/- SD (n = 3–4). Statistical analysis has been performed using ANOVA with Tukey’s post hoc test. ∗p < 0.001 and ∗∗p < 0.0001 compared with untreated control and #p < 0.01 and ##p < 0.001 from 2 h postglutamate time. COX-2, cyclooxygenase-2; p38 MAPK, p38 mitogen-activated protein kinase; STEP, striatal-enriched phosphatase. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34246631), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: STEP Antibody (23E5) - BSA Free [NB300-202] -

Role of NF-kappa B in glutamate-induced increase in the COX-2 protein level in STEP-deficient neurons.A and B, neuronal cultures from (A) WT and (B) STEP KO mice were treated with 50 μM glutamate (Glu) for 20 min and then maintained in their original medium for the specified times (post-Glu time). C–F, neuronal cultures from STEP KO mice were treated with 50 μM glutamate (Glu) for 20 min followed by recovery (post-Glu time) in the absence and presence of (C) MK801 (5 μM), (D) SB 203580 (5 μM), or (E and F) Bengamide B (500 nM). Equal amounts of protein from each sample were analyzed by immunoblotting using (A–D) anti-I kappaB alpha (top) and anti-beta -tubulin (bottom) antibodies, or (E) anti-COX-2 (top) and anti-beta -tubulin (bottom) antibodies. Corresponding bar diagrams represents quantitative analysis of (A–D) I kappaB alpha or (E) COX-2 protein levels as the mean +/- SD. F, equal amounts of culture media from each sample were analyzed for PGE2 level using ELISA. Statistical analysis has been performed using ANOVA with Tukey’s post hoc test. Values are expressed as the mean +/- SD (n = 3–4). ∗p < 0.01 and ∗∗p < 0.001 compared with the untreated control and #p < 0.01 from 2 h postglutamate time. COX-2, cyclooxygenase-2; I kappaB, inhibitor of nuclear factor-kappa B; STEP, striatal-enriched phosphatase. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34246631), licensed under a CC-BY license. Not internally tested by Novus Biologicals.