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catalytic subunit  (New England Biolabs)


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    Structured Review

    New England Biolabs catalytic subunit
    Catalytic Subunit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 772 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pka/cAMP-dependent+Protein+Kinase+(PKA)%2C+catalytic+subunit/10__1016_slash_j__snb__2026__141050-49-1-13
    Average 95 stars, based on 772 article reviews
    catalytic subunit - by Bioz Stars, 2026-11
    95/100 stars

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    Related Articles

    other:

    Article Title: Structural basis of CSN-mediated SCF deneddylation.
    Article Snippet: Data was collected using a Titan Krios cryo-EM (Thermo Fisher Scientific), operated at 300 kV, equipped with a Falcon III detector.

    Article Title: Structural Analysis of γ-phosphate modified ATP analogs Compatible with Kinase-Catalyzed Labeling
    Article Snippet: CKII and PKA were bought from New England Biolabs.

    Phospho-proteomics:

    Article Title: Tau condensation on DNA mediates microtubule attachment suggesting a mitotic role for centromere-localized tau.
    Article Snippet: The protein elutes were concentrated using a protein concentrator (Sartorius #VS15T01). .. Phosphorylation of wild-type tau was performed in vitro following Abasi et al.25 A total of 100 μM tau was incubated with 2 μL PKA (New England BioLabs #P6000S) in kinase buffer containing 20 mM HEPES (pH 7.2), 50 mM NaCl, 0.5 mM TCEP, 4 mM adenosine-5′-triphosphate (Sigma #A7699-5G), 1 mM PMSF and 10 mM MgCl2 at 30 °C for 16 h. To inactivate the kinase, the mixture was incubated at 65 °C for 20 min. ..

    Article Title: Tau condensation on DNA mediates microtubule attachment suggesting a mitotic role for centromere-localized tau
    Article Snippet: The protein elutes were concentrated using a protein concentrator (Sartorius #VS15T01). .. Phosphorylation of wild-type tau was performed in vitro following Abasi et al. . A total of 100 μM tau was incubated with 2 μL PKA (New England BioLabs #P6000S) in kinase buffer containing 20 mM HEPES (pH 7.2), 50 mM NaCl, 0.5 mM TCEP, 4 mM adenosine-5′-triphosphate (Sigma #A7699-5G), 1 mM PMSF and 10 mM MgCl 2 at 30 °C for 16 h. To inactivate the kinase, the mixture was incubated at 65 °C for 20 min. ..

    Article Title: Ultrasonic Reporter of Kinase Activity
    Article Snippet: .. For all phosphorylation assays, 250 units of PKA (P6000L; NEB; specific activity: 1 pmol/min/unit), 1.5 μg of human CaMKIIα (Sino Biological; specific activity: 160 nmol/min/mg), or 0.1 μg of PKC alpha (Abcam; specific activity: 3,200 nmol/min/mg) per μL of engineered GVs at OD500 = 10 were mixed with 1 mM ATP (Sigma-Aldrich) in NEBufferTM for Protein Kinases (50 mM Tris-HCl, 10 mM MgCl 2 , 0.1 mM EDTA, 2 mM DTT, 0.01% Brij 35, pH 7.5; NEB) and incubated with slow rotation at 37°C for 10–12 hours, unless otherwise specified. .. For dephosphorylation assays, 10 units of LPP (P0753L; NEB) per μL of engineered GVs at OD500 = 10 were mixed with PMP Buffer (50 mM HEPES, 100 mM NaCl, 2 mM DTT, 0.01% Brij 35, pH 7.5) and 1 mM MnCl 2 , followed by incubation with slow rotation at 37°C for 10–12 hours.

    In Vitro:

    Article Title: Tau condensation on DNA mediates microtubule attachment suggesting a mitotic role for centromere-localized tau.
    Article Snippet: The protein elutes were concentrated using a protein concentrator (Sartorius #VS15T01). .. Phosphorylation of wild-type tau was performed in vitro following Abasi et al.25 A total of 100 μM tau was incubated with 2 μL PKA (New England BioLabs #P6000S) in kinase buffer containing 20 mM HEPES (pH 7.2), 50 mM NaCl, 0.5 mM TCEP, 4 mM adenosine-5′-triphosphate (Sigma #A7699-5G), 1 mM PMSF and 10 mM MgCl2 at 30 °C for 16 h. To inactivate the kinase, the mixture was incubated at 65 °C for 20 min. ..

    Article Title: Tau condensation on DNA mediates microtubule attachment suggesting a mitotic role for centromere-localized tau
    Article Snippet: The protein elutes were concentrated using a protein concentrator (Sartorius #VS15T01). .. Phosphorylation of wild-type tau was performed in vitro following Abasi et al. . A total of 100 μM tau was incubated with 2 μL PKA (New England BioLabs #P6000S) in kinase buffer containing 20 mM HEPES (pH 7.2), 50 mM NaCl, 0.5 mM TCEP, 4 mM adenosine-5′-triphosphate (Sigma #A7699-5G), 1 mM PMSF and 10 mM MgCl 2 at 30 °C for 16 h. To inactivate the kinase, the mixture was incubated at 65 °C for 20 min. ..

    Article Title: PKA regulates stress granule maturation to allow timely recovery after prolonged starvation
    Article Snippet: .. In vitro kinase reactions were performed using recombinant Cdc19, which was mixed at a concentration of 0.5 mg/ml 1:50 with PKA (NEB, Cat# P6000S) in buffer containing 100 mM Tris-HCl pH7.5, 100 mM NaCl, 5% glycerol, 20 mM MgCl2, 3 mM DTT, 400 μM ATP (Sigma, Cat# A2383-5G), 1 mM PMSF with protease inhibitor tablets. ..

    Incubation:

    Article Title: Tau condensation on DNA mediates microtubule attachment suggesting a mitotic role for centromere-localized tau.
    Article Snippet: The protein elutes were concentrated using a protein concentrator (Sartorius #VS15T01). .. Phosphorylation of wild-type tau was performed in vitro following Abasi et al.25 A total of 100 μM tau was incubated with 2 μL PKA (New England BioLabs #P6000S) in kinase buffer containing 20 mM HEPES (pH 7.2), 50 mM NaCl, 0.5 mM TCEP, 4 mM adenosine-5′-triphosphate (Sigma #A7699-5G), 1 mM PMSF and 10 mM MgCl2 at 30 °C for 16 h. To inactivate the kinase, the mixture was incubated at 65 °C for 20 min. ..

    Article Title: Tau condensation on DNA mediates microtubule attachment suggesting a mitotic role for centromere-localized tau
    Article Snippet: The protein elutes were concentrated using a protein concentrator (Sartorius #VS15T01). .. Phosphorylation of wild-type tau was performed in vitro following Abasi et al. . A total of 100 μM tau was incubated with 2 μL PKA (New England BioLabs #P6000S) in kinase buffer containing 20 mM HEPES (pH 7.2), 50 mM NaCl, 0.5 mM TCEP, 4 mM adenosine-5′-triphosphate (Sigma #A7699-5G), 1 mM PMSF and 10 mM MgCl 2 at 30 °C for 16 h. To inactivate the kinase, the mixture was incubated at 65 °C for 20 min. ..

    Article Title: Ultrasonic Reporter of Kinase Activity
    Article Snippet: .. For all phosphorylation assays, 250 units of PKA (P6000L; NEB; specific activity: 1 pmol/min/unit), 1.5 μg of human CaMKIIα (Sino Biological; specific activity: 160 nmol/min/mg), or 0.1 μg of PKC alpha (Abcam; specific activity: 3,200 nmol/min/mg) per μL of engineered GVs at OD500 = 10 were mixed with 1 mM ATP (Sigma-Aldrich) in NEBufferTM for Protein Kinases (50 mM Tris-HCl, 10 mM MgCl 2 , 0.1 mM EDTA, 2 mM DTT, 0.01% Brij 35, pH 7.5; NEB) and incubated with slow rotation at 37°C for 10–12 hours, unless otherwise specified. .. For dephosphorylation assays, 10 units of LPP (P0753L; NEB) per μL of engineered GVs at OD500 = 10 were mixed with PMP Buffer (50 mM HEPES, 100 mM NaCl, 2 mM DTT, 0.01% Brij 35, pH 7.5) and 1 mM MnCl 2 , followed by incubation with slow rotation at 37°C for 10–12 hours.

    Activity Assay:

    Article Title: Ultrasonic Reporter of Kinase Activity
    Article Snippet: .. For all phosphorylation assays, 250 units of PKA (P6000L; NEB; specific activity: 1 pmol/min/unit), 1.5 μg of human CaMKIIα (Sino Biological; specific activity: 160 nmol/min/mg), or 0.1 μg of PKC alpha (Abcam; specific activity: 3,200 nmol/min/mg) per μL of engineered GVs at OD500 = 10 were mixed with 1 mM ATP (Sigma-Aldrich) in NEBufferTM for Protein Kinases (50 mM Tris-HCl, 10 mM MgCl 2 , 0.1 mM EDTA, 2 mM DTT, 0.01% Brij 35, pH 7.5; NEB) and incubated with slow rotation at 37°C for 10–12 hours, unless otherwise specified. .. For dephosphorylation assays, 10 units of LPP (P0753L; NEB) per μL of engineered GVs at OD500 = 10 were mixed with PMP Buffer (50 mM HEPES, 100 mM NaCl, 2 mM DTT, 0.01% Brij 35, pH 7.5) and 1 mM MnCl 2 , followed by incubation with slow rotation at 37°C for 10–12 hours.

    Recombinant:

    Article Title: PKA regulates stress granule maturation to allow timely recovery after prolonged starvation
    Article Snippet: .. In vitro kinase reactions were performed using recombinant Cdc19, which was mixed at a concentration of 0.5 mg/ml 1:50 with PKA (NEB, Cat# P6000S) in buffer containing 100 mM Tris-HCl pH7.5, 100 mM NaCl, 5% glycerol, 20 mM MgCl2, 3 mM DTT, 400 μM ATP (Sigma, Cat# A2383-5G), 1 mM PMSF with protease inhibitor tablets. ..

    Concentration Assay:

    Article Title: PKA regulates stress granule maturation to allow timely recovery after prolonged starvation
    Article Snippet: .. In vitro kinase reactions were performed using recombinant Cdc19, which was mixed at a concentration of 0.5 mg/ml 1:50 with PKA (NEB, Cat# P6000S) in buffer containing 100 mM Tris-HCl pH7.5, 100 mM NaCl, 5% glycerol, 20 mM MgCl2, 3 mM DTT, 400 μM ATP (Sigma, Cat# A2383-5G), 1 mM PMSF with protease inhibitor tablets. ..

    Protease Inhibitor:

    Article Title: PKA regulates stress granule maturation to allow timely recovery after prolonged starvation
    Article Snippet: .. In vitro kinase reactions were performed using recombinant Cdc19, which was mixed at a concentration of 0.5 mg/ml 1:50 with PKA (NEB, Cat# P6000S) in buffer containing 100 mM Tris-HCl pH7.5, 100 mM NaCl, 5% glycerol, 20 mM MgCl2, 3 mM DTT, 400 μM ATP (Sigma, Cat# A2383-5G), 1 mM PMSF with protease inhibitor tablets. ..



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    PKA inhibition by H89 abolishes the neuroprotective and antidepressant effects of NDP-MSH. (A) Experimental timeline and grouping strategy. Mice were assigned to Vehicle, LPS, LPS + NDP-MSH(NDP), LPS + NDP-MSH + H89(LNH), or H89 groups. (B) Representative locomotor traces in the OFT. (C) Behavioral assessments. NDP-MSH significantly increased sucrose preference in LPS-treated mice, whereas H89 co-treatment abolished this effect in the SPT. (One-way ANOVA, F (4, 41) = 13.96, p < 0.0001, followed by Tukey’s multiple comparisons test. Vehicle vs. LPS: q (41) = 6.649, p < 0.001; Vehicle vs. LNH: q (41) = 6.804, p < 0.001; LPS vs. NDP: q (41) = 4.128, p < 0.05; NDP vs. LNH: q (41) = 4.237, p < 0.05). NDP-MSH restored total distance traveled reduced by LPS; H89 partly reversed this improvement in the OFT (One-way ANOVA, F (4, 41) = 29.92, p < 0.0001, followed by Tukey’s multiple comparisons test. Vehicle vs. LPS: q (41) = 13.07, p < 0.0001; Vehicle vs. NDP: q (41) = 7.823, p < 0.0001; Vehicle vs. LNH: q (41) = 11.660, p < 0.0001; LPS vs. NDP, q (41) = 5.225, p < 0.01). NDP-MSH partly restored central zone entries reduced by LPS; H89 partly reversed this improvement in the OFT (Kruskal-Wallis non-parametric test, H (4) = 30.23, p < 0.0001, followed by Dunn's multiple comparisons test. Vehicle vs. LPS: Z = 4.745, p < 0.0001; Vehicle vs. NDP: Z = 2.600, p = 0.093; Vehicle vs. LNH: Z = 3.924, p < 0.0001; H89 vs. LPS: Z = 3.615, p < 0.01) (Veh, n = 8; H89, n = 8; LPS, n = 10; NDP, n = 9; LNH, n = 11). (D) ELISA quantification of IL-1β levels in the PFC. NDP-MSH reversed LPS-induced IL-1β elevation, whereas H89 prevented this effect (One-way ANOVA, F (4, 25) = 18.02, p < 0.0001, followed by Tukey’s multiple comparisons test. Vehicle vs. LPS, q (25) = 9.990, p < 0.0001; Vehicle vs. LNH: q (25) = 6.190, p < 0.01; LPS vs. NDP, q (25) = 8.350, p < 0.0001; NDP vs. LNH, q (25) = 4.549, p < 0.05) (n = 6 mice per group). (E) Representative images of mPFC neuronal dendrites by Golgi-Cox staining. Scale bar = 20 μm. Column graphs illustrate the relative spine numbers. NDP-MSH significantly increased spine density reduced by LPS treatment, whereas co-treatment with the PKA inhibitor H89 abolished this restorative effect (One-way ANOVA, F (4, 20) = 36.61, p < 0.0001, followed by Tukey’s multiple comparisons test. Veh vs. LPS, q (20) = 13.07, p < 0.0001; Veh vs. NDP, q (20) = 6.820, p < 0.001; Veh vs. LNH, q (20) = 12.82, p < 0.0001; LPS vs. NDP, q (20) = 6.245, p < 0.01; NDP vs. LNH, q (20) = 5.995, p < 0.01) (n = 5 mice per group, with 4-6 sections per mouse; at least 10 dendritic segments were randomly selected and quantified per section). (F) Quantification of relative Iba-1 fluorescence intensity, NDP-MSH normalized LPS-evoked microglial activation; H89 co-treatment restored microglial density to LPS-like levels (One-way ANOVA, F (4, 10) = 19.57, p < 0.001, followed by Tukey’s multiple comparisons test. Veh vs. LPS, q (10) = 8.650, p < 0.001; Veh vs. LNH, q (10) = 8.412, p < 0.01; LPS vs. NDP, q (10) = 5.940, p < 0.05; NDP vs. LNH, q (10) = 5.702, p < 0.05) (n = 3 mice per group). (G) Representative immunofluorescence images of Iba-1 staining in the mPFC, Scale bar = 100 µm. Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. ****p < 0.0001. Source: BioRender.com .

    Journal: Frontiers in Pharmacology

    Article Title: NDP-MSH rescues LPS-induced neuroinflammation, synaptic deficits, and depressive-like behaviors in mice: involvement of MC1R–cAMP/PKA signaling

    doi: 10.3389/fphar.2026.1895800

    Figure Lengend Snippet: PKA inhibition by H89 abolishes the neuroprotective and antidepressant effects of NDP-MSH. (A) Experimental timeline and grouping strategy. Mice were assigned to Vehicle, LPS, LPS + NDP-MSH(NDP), LPS + NDP-MSH + H89(LNH), or H89 groups. (B) Representative locomotor traces in the OFT. (C) Behavioral assessments. NDP-MSH significantly increased sucrose preference in LPS-treated mice, whereas H89 co-treatment abolished this effect in the SPT. (One-way ANOVA, F (4, 41) = 13.96, p < 0.0001, followed by Tukey’s multiple comparisons test. Vehicle vs. LPS: q (41) = 6.649, p < 0.001; Vehicle vs. LNH: q (41) = 6.804, p < 0.001; LPS vs. NDP: q (41) = 4.128, p < 0.05; NDP vs. LNH: q (41) = 4.237, p < 0.05). NDP-MSH restored total distance traveled reduced by LPS; H89 partly reversed this improvement in the OFT (One-way ANOVA, F (4, 41) = 29.92, p < 0.0001, followed by Tukey’s multiple comparisons test. Vehicle vs. LPS: q (41) = 13.07, p < 0.0001; Vehicle vs. NDP: q (41) = 7.823, p < 0.0001; Vehicle vs. LNH: q (41) = 11.660, p < 0.0001; LPS vs. NDP, q (41) = 5.225, p < 0.01). NDP-MSH partly restored central zone entries reduced by LPS; H89 partly reversed this improvement in the OFT (Kruskal-Wallis non-parametric test, H (4) = 30.23, p < 0.0001, followed by Dunn's multiple comparisons test. Vehicle vs. LPS: Z = 4.745, p < 0.0001; Vehicle vs. NDP: Z = 2.600, p = 0.093; Vehicle vs. LNH: Z = 3.924, p < 0.0001; H89 vs. LPS: Z = 3.615, p < 0.01) (Veh, n = 8; H89, n = 8; LPS, n = 10; NDP, n = 9; LNH, n = 11). (D) ELISA quantification of IL-1β levels in the PFC. NDP-MSH reversed LPS-induced IL-1β elevation, whereas H89 prevented this effect (One-way ANOVA, F (4, 25) = 18.02, p < 0.0001, followed by Tukey’s multiple comparisons test. Vehicle vs. LPS, q (25) = 9.990, p < 0.0001; Vehicle vs. LNH: q (25) = 6.190, p < 0.01; LPS vs. NDP, q (25) = 8.350, p < 0.0001; NDP vs. LNH, q (25) = 4.549, p < 0.05) (n = 6 mice per group). (E) Representative images of mPFC neuronal dendrites by Golgi-Cox staining. Scale bar = 20 μm. Column graphs illustrate the relative spine numbers. NDP-MSH significantly increased spine density reduced by LPS treatment, whereas co-treatment with the PKA inhibitor H89 abolished this restorative effect (One-way ANOVA, F (4, 20) = 36.61, p < 0.0001, followed by Tukey’s multiple comparisons test. Veh vs. LPS, q (20) = 13.07, p < 0.0001; Veh vs. NDP, q (20) = 6.820, p < 0.001; Veh vs. LNH, q (20) = 12.82, p < 0.0001; LPS vs. NDP, q (20) = 6.245, p < 0.01; NDP vs. LNH, q (20) = 5.995, p < 0.01) (n = 5 mice per group, with 4-6 sections per mouse; at least 10 dendritic segments were randomly selected and quantified per section). (F) Quantification of relative Iba-1 fluorescence intensity, NDP-MSH normalized LPS-evoked microglial activation; H89 co-treatment restored microglial density to LPS-like levels (One-way ANOVA, F (4, 10) = 19.57, p < 0.001, followed by Tukey’s multiple comparisons test. Veh vs. LPS, q (10) = 8.650, p < 0.001; Veh vs. LNH, q (10) = 8.412, p < 0.01; LPS vs. NDP, q (10) = 5.940, p < 0.05; NDP vs. LNH, q (10) = 5.702, p < 0.05) (n = 3 mice per group). (G) Representative immunofluorescence images of Iba-1 staining in the mPFC, Scale bar = 100 µm. Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. ****p < 0.0001. Source: BioRender.com .

    Article Snippet: Treatment regimens were as follows: vehicle-treated mice received equivalent volumes of saline; the LPS group received lipopolysaccharide (LPS; 1 mg/kg; L2880, Sigma-Aldrich, United States); the LPS + NDP-MSH group received LPS (1 mg/kg) together with NDP-MSH (2 mg/kg; HY-N2466, MedChemExpress, United States); the LPS + NDP-MSH + H89 group received LPS (1 mg/kg), NDP-MSH (2 mg/kg), and the PKA inhibitor H89 (10 mg/kg; HY-15979A, MedChemExpress, United States); and the H89-alone group received H89 (10 mg/kg).

    Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Staining, Fluorescence, Activation Assay, Immunofluorescence

    NDP-MSH Normalizes LPS-induced Metabolic Reprogramming through cAMP/PKA Signaling. (A) Principal component analysis (PCA) score plot of untargeted metabolomic profiles from saline control (CTRL), LPS-treated (LPS), drug-treated (NDP), drug plus inhibitor-treated (LNH), inhibitor alone (H89), and quality control (QC) samples. Biological samples included n = 4 mice per treatment group (CTRL, LPS, NDP, LNH, and H89). QC samples (n = 3) were used for quality control and were excluded from statistical comparisons. (B) Bar plot showing the number of significantly altered metabolites (upregulated and downregulated) across pairwise comparisons (limma, FDR <0.2, |log 2 fold change| ≥ 0.15; QC samples excluded). (C) Scatter plot showing the correlation between LPS-induced metabolic changes (LPS vs. CTRL) and drug-induced changes (NDP vs. LPS). (D) Scatter plot showing the correlation between drug-induced changes (NDP vs. LPS) and inhibitor-on-drug effects (LNH vs. NDP). (E) Heatmap showing a selected subset of metabolites whose LPS-induced changes were reversed by NDP-MSH treatment and whose NDP-MSH-mediated reversal was blocked by H89. Relative metabolite intensities were Z-score normalized across group means. (F) KEGG pathway enrichment analysis of drug-responsive metabolites. Source: BioRender.com .

    Journal: Frontiers in Pharmacology

    Article Title: NDP-MSH rescues LPS-induced neuroinflammation, synaptic deficits, and depressive-like behaviors in mice: involvement of MC1R–cAMP/PKA signaling

    doi: 10.3389/fphar.2026.1895800

    Figure Lengend Snippet: NDP-MSH Normalizes LPS-induced Metabolic Reprogramming through cAMP/PKA Signaling. (A) Principal component analysis (PCA) score plot of untargeted metabolomic profiles from saline control (CTRL), LPS-treated (LPS), drug-treated (NDP), drug plus inhibitor-treated (LNH), inhibitor alone (H89), and quality control (QC) samples. Biological samples included n = 4 mice per treatment group (CTRL, LPS, NDP, LNH, and H89). QC samples (n = 3) were used for quality control and were excluded from statistical comparisons. (B) Bar plot showing the number of significantly altered metabolites (upregulated and downregulated) across pairwise comparisons (limma, FDR <0.2, |log 2 fold change| ≥ 0.15; QC samples excluded). (C) Scatter plot showing the correlation between LPS-induced metabolic changes (LPS vs. CTRL) and drug-induced changes (NDP vs. LPS). (D) Scatter plot showing the correlation between drug-induced changes (NDP vs. LPS) and inhibitor-on-drug effects (LNH vs. NDP). (E) Heatmap showing a selected subset of metabolites whose LPS-induced changes were reversed by NDP-MSH treatment and whose NDP-MSH-mediated reversal was blocked by H89. Relative metabolite intensities were Z-score normalized across group means. (F) KEGG pathway enrichment analysis of drug-responsive metabolites. Source: BioRender.com .

    Article Snippet: Treatment regimens were as follows: vehicle-treated mice received equivalent volumes of saline; the LPS group received lipopolysaccharide (LPS; 1 mg/kg; L2880, Sigma-Aldrich, United States); the LPS + NDP-MSH group received LPS (1 mg/kg) together with NDP-MSH (2 mg/kg; HY-N2466, MedChemExpress, United States); the LPS + NDP-MSH + H89 group received LPS (1 mg/kg), NDP-MSH (2 mg/kg), and the PKA inhibitor H89 (10 mg/kg; HY-15979A, MedChemExpress, United States); and the H89-alone group received H89 (10 mg/kg).

    Techniques: Metabolomic, Saline, Control