adenylation Search Results


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  • 95
    New England Biolabs adenylation kit
    Adenylation Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Millipore adenylic acid
    JNK phosphorylates RARα and promotes ubiquitination. (A) JNK phosphorylates RARα in intact cells. HeLa cells were transiently cotransfected with expression vectors containing Flag-tagged RARα and JNK1, labeled with [ 32 P]orthophosphate, treated (+) or not treated (−) with UV radiation, and lysed. RARα was immunoprecipitated (IP) from cell lysates with anti-Flag antibodies, and the immunoprecipitate was visualized by autoradiography (top) or subjected to Western blotting with anti-RARα antibodies (bottom). (B) JNK phosphorylates RARα in vitro. JNK and p38 kinases were immunoprecipitated from HeLa cells after UV irradiation to activate stress pathways and subjected to in vitro kinase assays with GST-RARα, GST-c-Jun, and GST-ATF2 as substrates. GST-c-Jun and GST-ATF2 were positive controls for JNK and p38, respectively. (C) RARα and JNK1 physically interact in vivo. COS-1 cells were transiently cotransfected with expression vectors containing RARα and HA-tagged JNK1 or empty vector (−) and lysed, and the lysates were subjected to immunoprecipitation with RARα antibodies, followed by Western blotting with anti-HA and RAR antibodies. (D) RARα degradation is JNK dependent. GST or GST-RARα was incubated with (lanes 1, 2, 4, and 5) or without (lane 3) active JNK1. Afterwards, the mixture was subjected to an ubiquitination reaction in the presence of an S-100 fraction of HeLa extract, His-ubiquitin (lanes 1, 2, 3, and 5), and ATP (lanes 1 to 4). In lane 5, ATP was replaced by <t>AMP-PNP,</t> which supports ubiquitination but not kinase reactions.
    Adenylic Acid, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Bioo Scientific adenylation kit
    JNK phosphorylates RARα and promotes ubiquitination. (A) JNK phosphorylates RARα in intact cells. HeLa cells were transiently cotransfected with expression vectors containing Flag-tagged RARα and JNK1, labeled with [ 32 P]orthophosphate, treated (+) or not treated (−) with UV radiation, and lysed. RARα was immunoprecipitated (IP) from cell lysates with anti-Flag antibodies, and the immunoprecipitate was visualized by autoradiography (top) or subjected to Western blotting with anti-RARα antibodies (bottom). (B) JNK phosphorylates RARα in vitro. JNK and p38 kinases were immunoprecipitated from HeLa cells after UV irradiation to activate stress pathways and subjected to in vitro kinase assays with GST-RARα, GST-c-Jun, and GST-ATF2 as substrates. GST-c-Jun and GST-ATF2 were positive controls for JNK and p38, respectively. (C) RARα and JNK1 physically interact in vivo. COS-1 cells were transiently cotransfected with expression vectors containing RARα and HA-tagged JNK1 or empty vector (−) and lysed, and the lysates were subjected to immunoprecipitation with RARα antibodies, followed by Western blotting with anti-HA and RAR antibodies. (D) RARα degradation is JNK dependent. GST or GST-RARα was incubated with (lanes 1, 2, 4, and 5) or without (lane 3) active JNK1. Afterwards, the mixture was subjected to an ubiquitination reaction in the presence of an S-100 fraction of HeLa extract, His-ubiquitin (lanes 1, 2, 3, and 5), and ATP (lanes 1 to 4). In lane 5, ATP was replaced by <t>AMP-PNP,</t> which supports ubiquitination but not kinase reactions.
    Adenylation Kit, supplied by Bioo Scientific, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abcam adenylate cyclase
    A water soluble fraction of Smilax china L. leaf ethanol extract (wsSCLE) stimulates protein kinase A (PKA) signaling. (A) The expression of <t>adenylate</t> cyclase (AC) and PKA by wsSCLE treatment. Mature 3T3-L1 adipocytes were incubated with wsSCLE (0.1 and 0.25 mg/mL) for 24 h. The β-adrenergic receptors were induced (by Iso, an agonist) and inhibited (by Pro, an antagonist). (B) The induction of the phosphorylation (p) of PKA substrates (PKAs) by wsSCLE treatment. H89 treatment (10 µM) decreased wsSCLE-induced pPKAs. Data are expressed as mean ± SD of triplicate experiments, ## P
    Adenylate Cyclase, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Eurofins complete adenylation
    A water soluble fraction of Smilax china L. leaf ethanol extract (wsSCLE) stimulates protein kinase A (PKA) signaling. (A) The expression of <t>adenylate</t> cyclase (AC) and PKA by wsSCLE treatment. Mature 3T3-L1 adipocytes were incubated with wsSCLE (0.1 and 0.25 mg/mL) for 24 h. The β-adrenergic receptors were induced (by Iso, an agonist) and inhibited (by Pro, an antagonist). (B) The induction of the phosphorylation (p) of PKA substrates (PKAs) by wsSCLE treatment. H89 treatment (10 µM) decreased wsSCLE-induced pPKAs. Data are expressed as mean ± SD of triplicate experiments, ## P
    Complete Adenylation, supplied by Eurofins, used in various techniques. Bioz Stars score: 85/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Millipore adenylic acid deaminase
    A water soluble fraction of Smilax china L. leaf ethanol extract (wsSCLE) stimulates protein kinase A (PKA) signaling. (A) The expression of <t>adenylate</t> cyclase (AC) and PKA by wsSCLE treatment. Mature 3T3-L1 adipocytes were incubated with wsSCLE (0.1 and 0.25 mg/mL) for 24 h. The β-adrenergic receptors were induced (by Iso, an agonist) and inhibited (by Pro, an antagonist). (B) The induction of the phosphorylation (p) of PKA substrates (PKAs) by wsSCLE treatment. H89 treatment (10 µM) decreased wsSCLE-induced pPKAs. Data are expressed as mean ± SD of triplicate experiments, ## P
    Adenylic Acid Deaminase, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore adenylate kinase
    Enzymatic synthesis of isotopically labeled orotidines. [5'- 14 C]Orotidine is used as an example. [1'- 14 C]-, [1'- 3 H]-, [2'- 3 H]-, [4'- 3 H]-, [5'- 14 C]-, [5'- 3 H 2 ]-, [1, 3- 15 N 2 , 5'- 14 C]- and [3- 15 N, 5'- 14 C]Orotidine were prepared enzymatically from isotopically labeled riboses, glucoses and orotates through two-step procedure (see Experimental Methods for details). The enzymes used for the synthesis include ribokinase (RK), hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PDH), phosphogluconic acid dehydrogenase (PGDH), L-glutamic dehydrogenase (GDH), phosphoriboisomerase (PRI), <t>adenylate</t> kinase (AK), pyruvate kinase (PK), phospho-D-ribosyl-1-pyrophosphate synthase (PRPPase), orotate phosphoribosyltransferase (OPRT) and alkaline phosphatase (AP).
    Adenylate Kinase, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 93 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Promega adenylate tails
    Enzymatic synthesis of isotopically labeled orotidines. [5'- 14 C]Orotidine is used as an example. [1'- 14 C]-, [1'- 3 H]-, [2'- 3 H]-, [4'- 3 H]-, [5'- 14 C]-, [5'- 3 H 2 ]-, [1, 3- 15 N 2 , 5'- 14 C]- and [3- 15 N, 5'- 14 C]Orotidine were prepared enzymatically from isotopically labeled riboses, glucoses and orotates through two-step procedure (see Experimental Methods for details). The enzymes used for the synthesis include ribokinase (RK), hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PDH), phosphogluconic acid dehydrogenase (PGDH), L-glutamic dehydrogenase (GDH), phosphoriboisomerase (PRI), <t>adenylate</t> kinase (AK), pyruvate kinase (PK), phospho-D-ribosyl-1-pyrophosphate synthase (PRPPase), orotate phosphoribosyltransferase (OPRT) and alkaline phosphatase (AP).
    Adenylate Tails, supplied by Promega, used in various techniques. Bioz Stars score: 88/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology adenylate cyclase
    Enzymatic synthesis of isotopically labeled orotidines. [5'- 14 C]Orotidine is used as an example. [1'- 14 C]-, [1'- 3 H]-, [2'- 3 H]-, [4'- 3 H]-, [5'- 14 C]-, [5'- 3 H 2 ]-, [1, 3- 15 N 2 , 5'- 14 C]- and [3- 15 N, 5'- 14 C]Orotidine were prepared enzymatically from isotopically labeled riboses, glucoses and orotates through two-step procedure (see Experimental Methods for details). The enzymes used for the synthesis include ribokinase (RK), hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PDH), phosphogluconic acid dehydrogenase (PGDH), L-glutamic dehydrogenase (GDH), phosphoriboisomerase (PRI), <t>adenylate</t> kinase (AK), pyruvate kinase (PK), phospho-D-ribosyl-1-pyrophosphate synthase (PRPPase), orotate phosphoribosyltransferase (OPRT) and alkaline phosphatase (AP).
    Adenylate Cyclase, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Bioo Scientific air adenylated linker a
    Enzymatic synthesis of isotopically labeled orotidines. [5'- 14 C]Orotidine is used as an example. [1'- 14 C]-, [1'- 3 H]-, [2'- 3 H]-, [4'- 3 H]-, [5'- 14 C]-, [5'- 3 H 2 ]-, [1, 3- 15 N 2 , 5'- 14 C]- and [3- 15 N, 5'- 14 C]Orotidine were prepared enzymatically from isotopically labeled riboses, glucoses and orotates through two-step procedure (see Experimental Methods for details). The enzymes used for the synthesis include ribokinase (RK), hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PDH), phosphogluconic acid dehydrogenase (PGDH), L-glutamic dehydrogenase (GDH), phosphoriboisomerase (PRI), <t>adenylate</t> kinase (AK), pyruvate kinase (PK), phospho-D-ribosyl-1-pyrophosphate synthase (PRPPase), orotate phosphoribosyltransferase (OPRT) and alkaline phosphatase (AP).
    Air Adenylated Linker A, supplied by Bioo Scientific, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Millipore adenylate cyclase
    Enzymatic synthesis of isotopically labeled orotidines. [5'- 14 C]Orotidine is used as an example. [1'- 14 C]-, [1'- 3 H]-, [2'- 3 H]-, [4'- 3 H]-, [5'- 14 C]-, [5'- 3 H 2 ]-, [1, 3- 15 N 2 , 5'- 14 C]- and [3- 15 N, 5'- 14 C]Orotidine were prepared enzymatically from isotopically labeled riboses, glucoses and orotates through two-step procedure (see Experimental Methods for details). The enzymes used for the synthesis include ribokinase (RK), hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PDH), phosphogluconic acid dehydrogenase (PGDH), L-glutamic dehydrogenase (GDH), phosphoriboisomerase (PRI), <t>adenylate</t> kinase (AK), pyruvate kinase (PK), phospho-D-ribosyl-1-pyrophosphate synthase (PRPPase), orotate phosphoribosyltransferase (OPRT) and alkaline phosphatase (AP).
    Adenylate Cyclase, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Bioo Scientific adenylation
    Enzymatic synthesis of isotopically labeled orotidines. [5'- 14 C]Orotidine is used as an example. [1'- 14 C]-, [1'- 3 H]-, [2'- 3 H]-, [4'- 3 H]-, [5'- 14 C]-, [5'- 3 H 2 ]-, [1, 3- 15 N 2 , 5'- 14 C]- and [3- 15 N, 5'- 14 C]Orotidine were prepared enzymatically from isotopically labeled riboses, glucoses and orotates through two-step procedure (see Experimental Methods for details). The enzymes used for the synthesis include ribokinase (RK), hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PDH), phosphogluconic acid dehydrogenase (PGDH), L-glutamic dehydrogenase (GDH), phosphoriboisomerase (PRI), <t>adenylate</t> kinase (AK), pyruvate kinase (PK), phospho-D-ribosyl-1-pyrophosphate synthase (PRPPase), orotate phosphoribosyltransferase (OPRT) and alkaline phosphatase (AP).
    Adenylation, supplied by Bioo Scientific, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Illumina Inc adenylation
    A schematic representation of the sample preparation workflow. The processes of the TruSeq™ RNA Sample Preparation v2 low sample (LS) protocol (Illumina) performed manually and adopted for automated use with the SX-8G IP-Star® Compact System are illustrated. The automated protocol minimizes the hands-on time required for the error-prone manual steps of RNA-seq library synthesis, <t>adenylation,</t> and adaptor ligation including all related clean up steps and allows experimental multitasking for the researcher in task.
    Adenylation, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 94/100, based on 2052 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Mimetics ub adenylate mimetics
    A schematic representation of the sample preparation workflow. The processes of the TruSeq™ RNA Sample Preparation v2 low sample (LS) protocol (Illumina) performed manually and adopted for automated use with the SX-8G IP-Star® Compact System are illustrated. The automated protocol minimizes the hands-on time required for the error-prone manual steps of RNA-seq library synthesis, <t>adenylation,</t> and adaptor ligation including all related clean up steps and allows experimental multitasking for the researcher in task.
    Ub Adenylate Mimetics, supplied by Mimetics, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Illumina Inc adenylate
    A schematic representation of the sample preparation workflow. The processes of the TruSeq™ RNA Sample Preparation v2 low sample (LS) protocol (Illumina) performed manually and adopted for automated use with the SX-8G IP-Star® Compact System are illustrated. The automated protocol minimizes the hands-on time required for the error-prone manual steps of RNA-seq library synthesis, <t>adenylation,</t> and adaptor ligation including all related clean up steps and allows experimental multitasking for the researcher in task.
    Adenylate, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 93/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Millipore adenylate myo kinase
    A schematic representation of the sample preparation workflow. The processes of the TruSeq™ RNA Sample Preparation v2 low sample (LS) protocol (Illumina) performed manually and adopted for automated use with the SX-8G IP-Star® Compact System are illustrated. The automated protocol minimizes the hands-on time required for the error-prone manual steps of RNA-seq library synthesis, <t>adenylation,</t> and adaptor ligation including all related clean up steps and allows experimental multitasking for the researcher in task.
    Adenylate Myo Kinase, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    New England Biolabs adenylated
    A schematic representation of the sample preparation workflow. The processes of the TruSeq™ RNA Sample Preparation v2 low sample (LS) protocol (Illumina) performed manually and adopted for automated use with the SX-8G IP-Star® Compact System are illustrated. The automated protocol minimizes the hands-on time required for the error-prone manual steps of RNA-seq library synthesis, <t>adenylation,</t> and adaptor ligation including all related clean up steps and allows experimental multitasking for the researcher in task.
    Adenylated, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore pacap
    Epac2 is involved in <t>PACAP-induced</t> astrocytogenesis. (A) GFAP immunolabeling of neural precursor cells (NPCs) at day 0 (DAT0). NPCs from neither wild-type (WT) nor Epac2-knockout (KO) mice expressed GFAP at DAT0. (B) WT NPCs remarkably differentiated into astrocytes when cultured in <t>bFGF-free</t> media for 4 days (at DAT4), but KO NPCs showed much lower GFAP immunoreactivity than WT NPCs. (C) CNTF (50 ng/ml) induced differentiation of NPCs of both genotypes into astrocytes. (D) WT NPCs incubated in PACAP (100 nM)-treated media for 4 days expressed GFAP, but KO NPCs did not display astrocytic differentiation. (E) Ratio of GFAP-immunopositive cell number to total NPCs number. KO NPCs did not show GFAP-immunopositive astrocytes by DAT4 in bFGF-free media ( # P = 0.0001, independent-samples t -test, trial number = 8). Treatment with either CNTF or PACAP increased differentiation of WT NPCs into astrocytes compared to WT NPCs without treatment at DAT4 (*P = 0.003 for CNTF, *P = 0.002 for PACAP, independent-samples t -test, trial number in each experiment = 8), but KO NPCs did not induce astrocytic differentiation after PACAP treatment contrast to CNTF treatment ( ## P = 0.0002, independent-samples t -test, trial number = 8). (F) GFAP intensity of immunopositive cells. KO NPCs showed much lower intensity in GFAP immunoreactivity compared to WT NPCs at DAT4 in bFGF-free media ( # P = 0.0066, independent-samples t -test). Treatment with either CNTF or PACAP increased GFAP intensity in WT NPCs compared to WT NPCs without treatment at DAT4 (*P = 0.024 for CNTF, *P = 0.007 for PACAP, independent-samples t -test), but contrast to CNTF, KO NPCs did not increase GFAP immunoreactivity even after PACAP treatment compared to WT NPCs ( ## P = 0.00004, independent-samples t -test). Scale bar = 200 μm.
    Pacap, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Enzymatics adenylation
    Epac2 is involved in <t>PACAP-induced</t> astrocytogenesis. (A) GFAP immunolabeling of neural precursor cells (NPCs) at day 0 (DAT0). NPCs from neither wild-type (WT) nor Epac2-knockout (KO) mice expressed GFAP at DAT0. (B) WT NPCs remarkably differentiated into astrocytes when cultured in <t>bFGF-free</t> media for 4 days (at DAT4), but KO NPCs showed much lower GFAP immunoreactivity than WT NPCs. (C) CNTF (50 ng/ml) induced differentiation of NPCs of both genotypes into astrocytes. (D) WT NPCs incubated in PACAP (100 nM)-treated media for 4 days expressed GFAP, but KO NPCs did not display astrocytic differentiation. (E) Ratio of GFAP-immunopositive cell number to total NPCs number. KO NPCs did not show GFAP-immunopositive astrocytes by DAT4 in bFGF-free media ( # P = 0.0001, independent-samples t -test, trial number = 8). Treatment with either CNTF or PACAP increased differentiation of WT NPCs into astrocytes compared to WT NPCs without treatment at DAT4 (*P = 0.003 for CNTF, *P = 0.002 for PACAP, independent-samples t -test, trial number in each experiment = 8), but KO NPCs did not induce astrocytic differentiation after PACAP treatment contrast to CNTF treatment ( ## P = 0.0002, independent-samples t -test, trial number = 8). (F) GFAP intensity of immunopositive cells. KO NPCs showed much lower intensity in GFAP immunoreactivity compared to WT NPCs at DAT4 in bFGF-free media ( # P = 0.0066, independent-samples t -test). Treatment with either CNTF or PACAP increased GFAP intensity in WT NPCs compared to WT NPCs without treatment at DAT4 (*P = 0.024 for CNTF, *P = 0.007 for PACAP, independent-samples t -test), but contrast to CNTF, KO NPCs did not increase GFAP immunoreactivity even after PACAP treatment compared to WT NPCs ( ## P = 0.00004, independent-samples t -test). Scale bar = 200 μm.
    Adenylation, supplied by Enzymatics, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Cambrex adenylate nucleotide ratio assay
    Epac2 is involved in <t>PACAP-induced</t> astrocytogenesis. (A) GFAP immunolabeling of neural precursor cells (NPCs) at day 0 (DAT0). NPCs from neither wild-type (WT) nor Epac2-knockout (KO) mice expressed GFAP at DAT0. (B) WT NPCs remarkably differentiated into astrocytes when cultured in <t>bFGF-free</t> media for 4 days (at DAT4), but KO NPCs showed much lower GFAP immunoreactivity than WT NPCs. (C) CNTF (50 ng/ml) induced differentiation of NPCs of both genotypes into astrocytes. (D) WT NPCs incubated in PACAP (100 nM)-treated media for 4 days expressed GFAP, but KO NPCs did not display astrocytic differentiation. (E) Ratio of GFAP-immunopositive cell number to total NPCs number. KO NPCs did not show GFAP-immunopositive astrocytes by DAT4 in bFGF-free media ( # P = 0.0001, independent-samples t -test, trial number = 8). Treatment with either CNTF or PACAP increased differentiation of WT NPCs into astrocytes compared to WT NPCs without treatment at DAT4 (*P = 0.003 for CNTF, *P = 0.002 for PACAP, independent-samples t -test, trial number in each experiment = 8), but KO NPCs did not induce astrocytic differentiation after PACAP treatment contrast to CNTF treatment ( ## P = 0.0002, independent-samples t -test, trial number = 8). (F) GFAP intensity of immunopositive cells. KO NPCs showed much lower intensity in GFAP immunoreactivity compared to WT NPCs at DAT4 in bFGF-free media ( # P = 0.0066, independent-samples t -test). Treatment with either CNTF or PACAP increased GFAP intensity in WT NPCs compared to WT NPCs without treatment at DAT4 (*P = 0.024 for CNTF, *P = 0.007 for PACAP, independent-samples t -test), but contrast to CNTF, KO NPCs did not increase GFAP immunoreactivity even after PACAP treatment compared to WT NPCs ( ## P = 0.00004, independent-samples t -test). Scale bar = 200 μm.
    Adenylate Nucleotide Ratio Assay, supplied by Cambrex, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    JNK phosphorylates RARα and promotes ubiquitination. (A) JNK phosphorylates RARα in intact cells. HeLa cells were transiently cotransfected with expression vectors containing Flag-tagged RARα and JNK1, labeled with [ 32 P]orthophosphate, treated (+) or not treated (−) with UV radiation, and lysed. RARα was immunoprecipitated (IP) from cell lysates with anti-Flag antibodies, and the immunoprecipitate was visualized by autoradiography (top) or subjected to Western blotting with anti-RARα antibodies (bottom). (B) JNK phosphorylates RARα in vitro. JNK and p38 kinases were immunoprecipitated from HeLa cells after UV irradiation to activate stress pathways and subjected to in vitro kinase assays with GST-RARα, GST-c-Jun, and GST-ATF2 as substrates. GST-c-Jun and GST-ATF2 were positive controls for JNK and p38, respectively. (C) RARα and JNK1 physically interact in vivo. COS-1 cells were transiently cotransfected with expression vectors containing RARα and HA-tagged JNK1 or empty vector (−) and lysed, and the lysates were subjected to immunoprecipitation with RARα antibodies, followed by Western blotting with anti-HA and RAR antibodies. (D) RARα degradation is JNK dependent. GST or GST-RARα was incubated with (lanes 1, 2, 4, and 5) or without (lane 3) active JNK1. Afterwards, the mixture was subjected to an ubiquitination reaction in the presence of an S-100 fraction of HeLa extract, His-ubiquitin (lanes 1, 2, 3, and 5), and ATP (lanes 1 to 4). In lane 5, ATP was replaced by AMP-PNP, which supports ubiquitination but not kinase reactions.

    Journal: Molecular and Cellular Biology

    Article Title: c-Jun N-Terminal Kinase Contributes to Aberrant Retinoid Signaling in Lung Cancer Cells by Phosphorylating and Inducing Proteasomal Degradation of Retinoic Acid Receptor ?

    doi: 10.1128/MCB.25.3.1054-1069.2005

    Figure Lengend Snippet: JNK phosphorylates RARα and promotes ubiquitination. (A) JNK phosphorylates RARα in intact cells. HeLa cells were transiently cotransfected with expression vectors containing Flag-tagged RARα and JNK1, labeled with [ 32 P]orthophosphate, treated (+) or not treated (−) with UV radiation, and lysed. RARα was immunoprecipitated (IP) from cell lysates with anti-Flag antibodies, and the immunoprecipitate was visualized by autoradiography (top) or subjected to Western blotting with anti-RARα antibodies (bottom). (B) JNK phosphorylates RARα in vitro. JNK and p38 kinases were immunoprecipitated from HeLa cells after UV irradiation to activate stress pathways and subjected to in vitro kinase assays with GST-RARα, GST-c-Jun, and GST-ATF2 as substrates. GST-c-Jun and GST-ATF2 were positive controls for JNK and p38, respectively. (C) RARα and JNK1 physically interact in vivo. COS-1 cells were transiently cotransfected with expression vectors containing RARα and HA-tagged JNK1 or empty vector (−) and lysed, and the lysates were subjected to immunoprecipitation with RARα antibodies, followed by Western blotting with anti-HA and RAR antibodies. (D) RARα degradation is JNK dependent. GST or GST-RARα was incubated with (lanes 1, 2, 4, and 5) or without (lane 3) active JNK1. Afterwards, the mixture was subjected to an ubiquitination reaction in the presence of an S-100 fraction of HeLa extract, His-ubiquitin (lanes 1, 2, 3, and 5), and ATP (lanes 1 to 4). In lane 5, ATP was replaced by AMP-PNP, which supports ubiquitination but not kinase reactions.

    Article Snippet: We purchased and SB203580 (Calbiochem, San Diego, Calif.); His-ubiquitin, MG132, all- trans RA, 9- cis RA, E- 4-[2-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-2-naphthalenyl)-1-propenyl]benzoic acid (TTNPB), ATP, triphosphate 5′-adenylic acid (AMP-PNP), phorbol myristate acetate (PMA), insulin-like growth factor 1, creatine kinase, and phosphocreatine (Sigma-Aldrich, St. Louis, Mo.); lactacystin and Z-VAD-FMK (Biomol, Plymouth Meeting, Pa.); [γ-32 P]ATP, [32 P]orthophosphate, and [35 S]methionine (ICN Biomedicals, Inc., Costa Mesa, Calif.); polyclonal antibodies against human RARα, RARβ, RARγ, RXRα, MKK4, JNK, and poly-ADP ribose polymerase (Santa Cruz Biotechnology, Santa Cruz, Calif.); U0126 and polyclonal antibodies against human phospho-JNK (Thr183/Tyr185), phospho-c-Jun (Ser63), c-Jun, phospho-Erk (Thr202/Tyr204), Erk, phospho-Akt (Ser473), and Akt (Cell Signaling Technology, Beverly, Mass.); monoclonal antibodies against Flag epitope and actin (Sigma-Aldrich); anti-HA antibody (Roche Molecular Biochemicals, Indianapolis, Ind.); and purified recombinant glutathione S -transferase (GST)-ATF2, GST-c-Jun, and HSP27 (Upstate Biotechnology, Lake Placid, N.Y.).

    Techniques: Expressing, Labeling, Immunoprecipitation, Autoradiography, Western Blot, In Vitro, Irradiation, In Vivo, Plasmid Preparation, Incubation

    A water soluble fraction of Smilax china L. leaf ethanol extract (wsSCLE) stimulates protein kinase A (PKA) signaling. (A) The expression of adenylate cyclase (AC) and PKA by wsSCLE treatment. Mature 3T3-L1 adipocytes were incubated with wsSCLE (0.1 and 0.25 mg/mL) for 24 h. The β-adrenergic receptors were induced (by Iso, an agonist) and inhibited (by Pro, an antagonist). (B) The induction of the phosphorylation (p) of PKA substrates (PKAs) by wsSCLE treatment. H89 treatment (10 µM) decreased wsSCLE-induced pPKAs. Data are expressed as mean ± SD of triplicate experiments, ## P

    Journal: Nutrition Research and Practice

    Article Title: Antiobesity effects of the water-soluble fraction of the ethanol extract of Smilax china L. leaf in 3T3-L1 adipocytes

    doi: 10.4162/nrp.2015.9.6.606

    Figure Lengend Snippet: A water soluble fraction of Smilax china L. leaf ethanol extract (wsSCLE) stimulates protein kinase A (PKA) signaling. (A) The expression of adenylate cyclase (AC) and PKA by wsSCLE treatment. Mature 3T3-L1 adipocytes were incubated with wsSCLE (0.1 and 0.25 mg/mL) for 24 h. The β-adrenergic receptors were induced (by Iso, an agonist) and inhibited (by Pro, an antagonist). (B) The induction of the phosphorylation (p) of PKA substrates (PKAs) by wsSCLE treatment. H89 treatment (10 µM) decreased wsSCLE-induced pPKAs. Data are expressed as mean ± SD of triplicate experiments, ## P

    Article Snippet: The membrane was placed in blocking buffer consisting of tris-buffered saline (TBS) supplemented with 0.1% tween 20 and 5% skim milk and was incubated for 2 h. The membrane was subsequently incubated in buffer containing primary antibody for 2 h. The antibodies, and dilutions used, were as follows: adenylate cyclase (1000:1) (Abcam, Cambridge, UK); PKA (2500:1); p-PKA (2000:1); p-PKA substrate (2000:1); hormone-sensitive lipase (HSL; 3000:1); GAPDH (2000:1; Santa Cruz Biotechnology, Santa Cruz, CA, USA); p-HSL (Ser563; 1000:1);and p-HSL (Ser660; 1000:1; Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Expressing, Incubation

    Enzymatic synthesis of isotopically labeled orotidines. [5'- 14 C]Orotidine is used as an example. [1'- 14 C]-, [1'- 3 H]-, [2'- 3 H]-, [4'- 3 H]-, [5'- 14 C]-, [5'- 3 H 2 ]-, [1, 3- 15 N 2 , 5'- 14 C]- and [3- 15 N, 5'- 14 C]Orotidine were prepared enzymatically from isotopically labeled riboses, glucoses and orotates through two-step procedure (see Experimental Methods for details). The enzymes used for the synthesis include ribokinase (RK), hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PDH), phosphogluconic acid dehydrogenase (PGDH), L-glutamic dehydrogenase (GDH), phosphoriboisomerase (PRI), adenylate kinase (AK), pyruvate kinase (PK), phospho-D-ribosyl-1-pyrophosphate synthase (PRPPase), orotate phosphoribosyltransferase (OPRT) and alkaline phosphatase (AP).

    Journal: Journal of the American Chemical Society

    Article Title: Pyrophosphate Interactions at the Transition States of Plasmodium falciparum and Human Orotate Phosphoribosyltransferases

    doi: 10.1021/ja102849w

    Figure Lengend Snippet: Enzymatic synthesis of isotopically labeled orotidines. [5'- 14 C]Orotidine is used as an example. [1'- 14 C]-, [1'- 3 H]-, [2'- 3 H]-, [4'- 3 H]-, [5'- 14 C]-, [5'- 3 H 2 ]-, [1, 3- 15 N 2 , 5'- 14 C]- and [3- 15 N, 5'- 14 C]Orotidine were prepared enzymatically from isotopically labeled riboses, glucoses and orotates through two-step procedure (see Experimental Methods for details). The enzymes used for the synthesis include ribokinase (RK), hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PDH), phosphogluconic acid dehydrogenase (PGDH), L-glutamic dehydrogenase (GDH), phosphoriboisomerase (PRI), adenylate kinase (AK), pyruvate kinase (PK), phospho-D-ribosyl-1-pyrophosphate synthase (PRPPase), orotate phosphoribosyltransferase (OPRT) and alkaline phosphatase (AP).

    Article Snippet: Hexokinase (HK), glucose 6-phosphate dehydrogenase (G6PDH), phosphogluconic acid dehydrogenase (PGDH), L-glutamic acid dehydrogenase (GDH), phosphoriboisomerase (PRI), adenylate kinase (AK) and pyruvate kinase (PK) were purchased from Sigma-Aldrich.

    Techniques: Labeling

    A schematic representation of the sample preparation workflow. The processes of the TruSeq™ RNA Sample Preparation v2 low sample (LS) protocol (Illumina) performed manually and adopted for automated use with the SX-8G IP-Star® Compact System are illustrated. The automated protocol minimizes the hands-on time required for the error-prone manual steps of RNA-seq library synthesis, adenylation, and adaptor ligation including all related clean up steps and allows experimental multitasking for the researcher in task.

    Journal: BMC Research Notes

    Article Title: An automated method for efficient, accurate and reproducible construction of RNA-seq libraries

    doi: 10.1186/s13104-015-1089-9

    Figure Lengend Snippet: A schematic representation of the sample preparation workflow. The processes of the TruSeq™ RNA Sample Preparation v2 low sample (LS) protocol (Illumina) performed manually and adopted for automated use with the SX-8G IP-Star® Compact System are illustrated. The automated protocol minimizes the hands-on time required for the error-prone manual steps of RNA-seq library synthesis, adenylation, and adaptor ligation including all related clean up steps and allows experimental multitasking for the researcher in task.

    Article Snippet: Use of the SX-8G IP-Star® Compact System significantly reduced the hands-on time for RNA-seq library synthesis, adenylation, and adaptor ligation providing with high quality RNA-seq libraries tailored for Illumina high-throughput next-generation sequencing.

    Techniques: Sample Prep, RNA Sequencing Assay, Ligation

    Epac2 is involved in PACAP-induced astrocytogenesis. (A) GFAP immunolabeling of neural precursor cells (NPCs) at day 0 (DAT0). NPCs from neither wild-type (WT) nor Epac2-knockout (KO) mice expressed GFAP at DAT0. (B) WT NPCs remarkably differentiated into astrocytes when cultured in bFGF-free media for 4 days (at DAT4), but KO NPCs showed much lower GFAP immunoreactivity than WT NPCs. (C) CNTF (50 ng/ml) induced differentiation of NPCs of both genotypes into astrocytes. (D) WT NPCs incubated in PACAP (100 nM)-treated media for 4 days expressed GFAP, but KO NPCs did not display astrocytic differentiation. (E) Ratio of GFAP-immunopositive cell number to total NPCs number. KO NPCs did not show GFAP-immunopositive astrocytes by DAT4 in bFGF-free media ( # P = 0.0001, independent-samples t -test, trial number = 8). Treatment with either CNTF or PACAP increased differentiation of WT NPCs into astrocytes compared to WT NPCs without treatment at DAT4 (*P = 0.003 for CNTF, *P = 0.002 for PACAP, independent-samples t -test, trial number in each experiment = 8), but KO NPCs did not induce astrocytic differentiation after PACAP treatment contrast to CNTF treatment ( ## P = 0.0002, independent-samples t -test, trial number = 8). (F) GFAP intensity of immunopositive cells. KO NPCs showed much lower intensity in GFAP immunoreactivity compared to WT NPCs at DAT4 in bFGF-free media ( # P = 0.0066, independent-samples t -test). Treatment with either CNTF or PACAP increased GFAP intensity in WT NPCs compared to WT NPCs without treatment at DAT4 (*P = 0.024 for CNTF, *P = 0.007 for PACAP, independent-samples t -test), but contrast to CNTF, KO NPCs did not increase GFAP immunoreactivity even after PACAP treatment compared to WT NPCs ( ## P = 0.00004, independent-samples t -test). Scale bar = 200 μm.

    Journal: BMB Reports

    Article Title: Epac2 contributes to PACAP-induced astrocytic differentiation through calcium ion influx in neural precursor cells

    doi: 10.5483/BMBRep.2016.49.2.202

    Figure Lengend Snippet: Epac2 is involved in PACAP-induced astrocytogenesis. (A) GFAP immunolabeling of neural precursor cells (NPCs) at day 0 (DAT0). NPCs from neither wild-type (WT) nor Epac2-knockout (KO) mice expressed GFAP at DAT0. (B) WT NPCs remarkably differentiated into astrocytes when cultured in bFGF-free media for 4 days (at DAT4), but KO NPCs showed much lower GFAP immunoreactivity than WT NPCs. (C) CNTF (50 ng/ml) induced differentiation of NPCs of both genotypes into astrocytes. (D) WT NPCs incubated in PACAP (100 nM)-treated media for 4 days expressed GFAP, but KO NPCs did not display astrocytic differentiation. (E) Ratio of GFAP-immunopositive cell number to total NPCs number. KO NPCs did not show GFAP-immunopositive astrocytes by DAT4 in bFGF-free media ( # P = 0.0001, independent-samples t -test, trial number = 8). Treatment with either CNTF or PACAP increased differentiation of WT NPCs into astrocytes compared to WT NPCs without treatment at DAT4 (*P = 0.003 for CNTF, *P = 0.002 for PACAP, independent-samples t -test, trial number in each experiment = 8), but KO NPCs did not induce astrocytic differentiation after PACAP treatment contrast to CNTF treatment ( ## P = 0.0002, independent-samples t -test, trial number = 8). (F) GFAP intensity of immunopositive cells. KO NPCs showed much lower intensity in GFAP immunoreactivity compared to WT NPCs at DAT4 in bFGF-free media ( # P = 0.0066, independent-samples t -test). Treatment with either CNTF or PACAP increased GFAP intensity in WT NPCs compared to WT NPCs without treatment at DAT4 (*P = 0.024 for CNTF, *P = 0.007 for PACAP, independent-samples t -test), but contrast to CNTF, KO NPCs did not increase GFAP immunoreactivity even after PACAP treatment compared to WT NPCs ( ## P = 0.00004, independent-samples t -test). Scale bar = 200 μm.

    Article Snippet: For astrocyte differentiation, cells were cultured in DMEM with N2 or calcium-free DMEM (Gibco, Thermo Fisher, Waltham, MA) with N2, which was then replaced with bFGF-free DMEM with N2 and a differentiation factor, such as 100 nM PACAP (A1439, Sigma- Aldrich, St Louis, MO) or 50 ng/ml CNTF (1203158, PeproTech) for 4 days.

    Techniques: Immunolabeling, Knock-Out, Mouse Assay, Cell Culture, Incubation