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Cell Signaling Technology Inc kinase buffer
Kinase Buffer, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 101 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/kinase buffer/product/Cell Signaling Technology Inc
Average 99 stars, based on 101 article reviews
Price from $9.99 to $1999.99
kinase buffer - by Bioz Stars, 2020-09
99/100 stars

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In Vitro:

Article Title: Proto-Oncogenic Src Phosphorylates EB1 to Regulate the Microtubule-Focal Adhesion Crosstalk and Stimulate Cell Migration
Article Snippet: .. In vitro kinase assay GFP-Src immunoprecipitated from HEK293T cells or bacterially purified GST-Src was incubated with bacterially purified His-EB1 at 30°C for 2.5 h in the kinase reaction buffer (Cell Signaling Technology) containing ATP (200 µM). ..

Article Title: Identification of a novel AMPK-PEA15 axis in the anoikis-resistant growth of mammary cells
Article Snippet: .. For radioactive in vitro kinase assay, 1 μg of purified PEA15 was incubated in the presence or absence of 1 μg of AMPK heterotrimer along with (γ32P) ATP (0.3 μCi) for 30 min at 30°C in AMPK kinase buffer (Cell Signaling Technology) supplemented with 50 μM ATP and 0.1 mM AMP. ..

Purification:

Article Title: Proto-Oncogenic Src Phosphorylates EB1 to Regulate the Microtubule-Focal Adhesion Crosstalk and Stimulate Cell Migration
Article Snippet: .. In vitro kinase assay GFP-Src immunoprecipitated from HEK293T cells or bacterially purified GST-Src was incubated with bacterially purified His-EB1 at 30°C for 2.5 h in the kinase reaction buffer (Cell Signaling Technology) containing ATP (200 µM). ..

Article Title: Identification of a novel AMPK-PEA15 axis in the anoikis-resistant growth of mammary cells
Article Snippet: .. For radioactive in vitro kinase assay, 1 μg of purified PEA15 was incubated in the presence or absence of 1 μg of AMPK heterotrimer along with (γ32P) ATP (0.3 μCi) for 30 min at 30°C in AMPK kinase buffer (Cell Signaling Technology) supplemented with 50 μM ATP and 0.1 mM AMP. ..

Immunoprecipitation:

Article Title: Proto-Oncogenic Src Phosphorylates EB1 to Regulate the Microtubule-Focal Adhesion Crosstalk and Stimulate Cell Migration
Article Snippet: .. In vitro kinase assay GFP-Src immunoprecipitated from HEK293T cells or bacterially purified GST-Src was incubated with bacterially purified His-EB1 at 30°C for 2.5 h in the kinase reaction buffer (Cell Signaling Technology) containing ATP (200 µM). ..

Article Title: Drosophila homeodomain-interacting protein kinase inhibits the Skp1-Cul1-F-box E3 ligase complex to dually promote Wingless and Hedgehog signaling
Article Snippet: .. Kinase assays were performed by using kinase assay buffer/ATP (Cell Signaling Technology) with GST-Hipk and other proteins immunoprecipitated from Drosophila S2 cells. .. Kinase assay reactions were incubated at 30 °C for 30 min followed by SDS/PAGE/Western blotting and detection with a phospho-specific β-catenin (S33/S37) antibody (1:750) (Cell Signaling Technology) or autoradiography.

Incubation:

Article Title: Proto-Oncogenic Src Phosphorylates EB1 to Regulate the Microtubule-Focal Adhesion Crosstalk and Stimulate Cell Migration
Article Snippet: .. In vitro kinase assay GFP-Src immunoprecipitated from HEK293T cells or bacterially purified GST-Src was incubated with bacterially purified His-EB1 at 30°C for 2.5 h in the kinase reaction buffer (Cell Signaling Technology) containing ATP (200 µM). ..

Article Title: Identification of a novel AMPK-PEA15 axis in the anoikis-resistant growth of mammary cells
Article Snippet: .. For radioactive in vitro kinase assay, 1 μg of purified PEA15 was incubated in the presence or absence of 1 μg of AMPK heterotrimer along with (γ32P) ATP (0.3 μCi) for 30 min at 30°C in AMPK kinase buffer (Cell Signaling Technology) supplemented with 50 μM ATP and 0.1 mM AMP. ..

Article Title: Synergistic Neuroprotective Effects of Lithium and Valproic Acid or Other Histone Deacetylase Inhibitors in Neurons: Roles of Glycogen Synthase Kinase-3 Inhibition
Article Snippet: .. The immunocomplex was bound to protein G Sepharose (GE Healthcare) by incubation at 4°C for 2 h and then washed three times with kinase assay buffer (Cell Signaling Technology). .. Phosphorylation of a typical GSK-3 β substrate (Cell Signaling Technology) by the kinase was performed by incubation for 30 min at 37°C in 40 μ l of reaction mixture, containing 2 μ g of substrate, 30 mM Tris-HCl, pH 7.5, 1 μ Ci [ γ -32 P]ATP, 4 mM MgCl2 , 2.5 mM β -glycerophosphate, 1 mM DTT, 0.05 mM Na3 VO4 , 40 μ g/ml bovine serum albumin, 100 μ M ATP, and GSK-3 β immunocomplex in the absence or presence of 3 mM LiCl, 0.8 mM VPA (sodium salt), or a combination of lithium and VPA.

Article Title: CDK5-mediated phosphorylation and stabilization of TPX2 promotes hepatocellular tumorigenesis
Article Snippet: .. A substrate (1 μg) was added into kinase assay buffer (CST) containing 25 mM Tris-HCl (pH 7.5), 2 mM dithiothreitol (DTT), 5 mM beta-glycerophosphate, 0.1 mM Na3 VO4 , and 10 mM MgCl2 , and incubated with CDK5/p25 kinase and 50 μM ATP-γ-S at 30 °C for 45 min. .. The samples were alkylated with 2.5 mM PNBM/5% DMSO (Abcam), incubated at room temperature for 1 h, and then subjected to western blotting.

Article Title: RSK2 Protein Suppresses Integrin Activation and Fibronectin Matrix Assembly and Promotes Cell Migration *
Article Snippet: .. Briefly, 10 μg of substrate was incubated with 100 ng of active RSK2 (SignalChem), and 1.25 nmol of ATP in 1× kinase buffer (Cell Signaling) or 0.8 μCi/μl [γ-32 P]ATP (PerkinElmer Life Sciences) with 1.25 nmol of ATP in 3× kinase buffer. ..

Activity Assay:

Article Title: p19Ink4d Is a Tumor Suppressor and Controls Pituitary Anterior Lobe Cell Proliferation
Article Snippet: .. Alternatively, for detection of kinase activity by Western blotting, Laemmli sample buffer was added to the completed kinase reaction mixture and a phosphothreonine-specific antibody (Cell Signaling) was used to detect phosphorylated Rb. .. We deleted the mouse p19 gene by targeted homologous recombination.

Western Blot:

Article Title: p19Ink4d Is a Tumor Suppressor and Controls Pituitary Anterior Lobe Cell Proliferation
Article Snippet: .. Alternatively, for detection of kinase activity by Western blotting, Laemmli sample buffer was added to the completed kinase reaction mixture and a phosphothreonine-specific antibody (Cell Signaling) was used to detect phosphorylated Rb. .. We deleted the mouse p19 gene by targeted homologous recombination.

Kinase Assay:

Article Title: Proto-Oncogenic Src Phosphorylates EB1 to Regulate the Microtubule-Focal Adhesion Crosstalk and Stimulate Cell Migration
Article Snippet: .. In vitro kinase assay GFP-Src immunoprecipitated from HEK293T cells or bacterially purified GST-Src was incubated with bacterially purified His-EB1 at 30°C for 2.5 h in the kinase reaction buffer (Cell Signaling Technology) containing ATP (200 µM). ..

Article Title: Identification of a novel AMPK-PEA15 axis in the anoikis-resistant growth of mammary cells
Article Snippet: .. For radioactive in vitro kinase assay, 1 μg of purified PEA15 was incubated in the presence or absence of 1 μg of AMPK heterotrimer along with (γ32P) ATP (0.3 μCi) for 30 min at 30°C in AMPK kinase buffer (Cell Signaling Technology) supplemented with 50 μM ATP and 0.1 mM AMP. ..

Article Title: Synergistic Neuroprotective Effects of Lithium and Valproic Acid or Other Histone Deacetylase Inhibitors in Neurons: Roles of Glycogen Synthase Kinase-3 Inhibition
Article Snippet: .. The immunocomplex was bound to protein G Sepharose (GE Healthcare) by incubation at 4°C for 2 h and then washed three times with kinase assay buffer (Cell Signaling Technology). .. Phosphorylation of a typical GSK-3 β substrate (Cell Signaling Technology) by the kinase was performed by incubation for 30 min at 37°C in 40 μ l of reaction mixture, containing 2 μ g of substrate, 30 mM Tris-HCl, pH 7.5, 1 μ Ci [ γ -32 P]ATP, 4 mM MgCl2 , 2.5 mM β -glycerophosphate, 1 mM DTT, 0.05 mM Na3 VO4 , 40 μ g/ml bovine serum albumin, 100 μ M ATP, and GSK-3 β immunocomplex in the absence or presence of 3 mM LiCl, 0.8 mM VPA (sodium salt), or a combination of lithium and VPA.

Article Title: Drosophila homeodomain-interacting protein kinase inhibits the Skp1-Cul1-F-box E3 ligase complex to dually promote Wingless and Hedgehog signaling
Article Snippet: .. Kinase assays were performed by using kinase assay buffer/ATP (Cell Signaling Technology) with GST-Hipk and other proteins immunoprecipitated from Drosophila S2 cells. .. Kinase assay reactions were incubated at 30 °C for 30 min followed by SDS/PAGE/Western blotting and detection with a phospho-specific β-catenin (S33/S37) antibody (1:750) (Cell Signaling Technology) or autoradiography.

Article Title: CDK5-mediated phosphorylation and stabilization of TPX2 promotes hepatocellular tumorigenesis
Article Snippet: .. A substrate (1 μg) was added into kinase assay buffer (CST) containing 25 mM Tris-HCl (pH 7.5), 2 mM dithiothreitol (DTT), 5 mM beta-glycerophosphate, 0.1 mM Na3 VO4 , and 10 mM MgCl2 , and incubated with CDK5/p25 kinase and 50 μM ATP-γ-S at 30 °C for 45 min. .. The samples were alkylated with 2.5 mM PNBM/5% DMSO (Abcam), incubated at room temperature for 1 h, and then subjected to western blotting.

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    Cell Signaling Technology Inc in vitro kinase assay buffer
    CDK5 Directly Binds to and Phos-phorylates to Activate CREB1 <t>in</t> GSCs (A) CP681301 treatment of mice xenografted explant culture tumors induced a clear down-regulation of CREB1 phosphorylation. (B) Western blot data using these tumor tissues also show downregulation of CREB1 phosphorylation. (C) We then investigated the mechanism by which CDK5 regulates CREB1. We found that Cdk5 can directly bind with CREB1, and CP681301 reduces the binding of CDK5 to CREB1. (D) An in <t>vitro</t> <t>kinase</t> <t>assay</t> showed that CDK5 can phosphorylate CREB1 at serine 133, and CP681301 can suppress this phosphorylation in a dose-dependent manner. (E) Diagram showing our proposed model where CDK5 directly phosphorylates and activates CREB1 without the requirement of cAMP/PKA in GSCs.
    In Vitro Kinase Assay Buffer, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/in vitro kinase assay buffer/product/Cell Signaling Technology Inc
    Average 99 stars, based on 21 article reviews
    Price from $9.99 to $1999.99
    in vitro kinase assay buffer - by Bioz Stars, 2020-09
    99/100 stars
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    CDK5 Directly Binds to and Phos-phorylates to Activate CREB1 in GSCs (A) CP681301 treatment of mice xenografted explant culture tumors induced a clear down-regulation of CREB1 phosphorylation. (B) Western blot data using these tumor tissues also show downregulation of CREB1 phosphorylation. (C) We then investigated the mechanism by which CDK5 regulates CREB1. We found that Cdk5 can directly bind with CREB1, and CP681301 reduces the binding of CDK5 to CREB1. (D) An in vitro kinase assay showed that CDK5 can phosphorylate CREB1 at serine 133, and CP681301 can suppress this phosphorylation in a dose-dependent manner. (E) Diagram showing our proposed model where CDK5 directly phosphorylates and activates CREB1 without the requirement of cAMP/PKA in GSCs.

    Journal: Cell reports

    Article Title: CDK5 Inhibition Resolves PKA/cAMP-Independent Activation of CREB1 Signaling in Glioma Stem Cells

    doi: 10.1016/j.celrep.2018.04.016

    Figure Lengend Snippet: CDK5 Directly Binds to and Phos-phorylates to Activate CREB1 in GSCs (A) CP681301 treatment of mice xenografted explant culture tumors induced a clear down-regulation of CREB1 phosphorylation. (B) Western blot data using these tumor tissues also show downregulation of CREB1 phosphorylation. (C) We then investigated the mechanism by which CDK5 regulates CREB1. We found that Cdk5 can directly bind with CREB1, and CP681301 reduces the binding of CDK5 to CREB1. (D) An in vitro kinase assay showed that CDK5 can phosphorylate CREB1 at serine 133, and CP681301 can suppress this phosphorylation in a dose-dependent manner. (E) Diagram showing our proposed model where CDK5 directly phosphorylates and activates CREB1 without the requirement of cAMP/PKA in GSCs.

    Article Snippet: The in vitro kinase assay buffer, ATP (Cell Signaling Tech), and 0, 0.5 μM, and 1 μM of CP681301 were added and mixed well.

    Techniques: Mouse Assay, Western Blot, Binding Assay, In Vitro, Kinase Assay

    Phosphoprotein enriched in astrocytes 15 kDa (PEA15) is a novel AMP-activated protein kinase (AMPK) substrate. (A) Primary human mammary epithelial cells (HMECs) cultured in ultra-low (UL) plates were transfected with control small interfering RNA (siRNA) or siRNA targeting PEA15. Two days posttransfection, a quarter of the cells were harvested and subjected to immunoblot analysis for the specified proteins (i), while the rest were scored for total number of mammospheres (MS) formed at the end of a week (ii). Error bars represent standard error of the mean (SEM); n = 4. (B) Primary HMECs cultured in UL plates were infected with control (empty vector) CSCG lentivirus, or those encoding for PEA15-WT or PEA15-S116A mutant with similar infection efficiencies (as scored by GFP-positive cells). Two days following infection, 1 × 10 5 transduced cells were seeded in methylcellulose and total number of GFP-positive mammospheres formed was counted at the end of a week. Error bars represent standard error of the mean (SEM); n = 3. (C) Primary HMECs seeded in UL plates were treated with AMPK activator A769962 (100 μM) for 0 min, 30 min, 1 hr and 2 hrs, and subjected to immunoblot analysis for the specified proteins; n = 3. (D) Primary HMECs seeded in UL plates were subjected to immunoprecipitation with total AMPK antibody followed by immunoblotting for specified proteins. Rabbit immunoglobulin G (IgG)-treated lysate and resin alone served as controls, n = 3. (E) Commercially procured heterotrimeric AMPK and pure PEA15 were incubated in an in vitro kinase assay buffer in the presence of 50 μM adenosine triphosphate (ATP) and 0.1 mM AMP for 30 min, and then subjected to immunoblot analyses with commercially available antibodies that specifically recognize PEA15 Ser 116 phosphorylation; total PEA15 antibody served as a loading control. (F) Autoradiography of in vitro kinase assay using pure AMPK and PEA15 proteins performed in the presence of γ 32 P-labeled ATP (0.3 μCi) for 30 min; n = 3.

    Journal: Breast Cancer Research : BCR

    Article Title: Identification of a novel AMPK-PEA15 axis in the anoikis-resistant growth of mammary cells

    doi: 10.1186/s13058-014-0420-z

    Figure Lengend Snippet: Phosphoprotein enriched in astrocytes 15 kDa (PEA15) is a novel AMP-activated protein kinase (AMPK) substrate. (A) Primary human mammary epithelial cells (HMECs) cultured in ultra-low (UL) plates were transfected with control small interfering RNA (siRNA) or siRNA targeting PEA15. Two days posttransfection, a quarter of the cells were harvested and subjected to immunoblot analysis for the specified proteins (i), while the rest were scored for total number of mammospheres (MS) formed at the end of a week (ii). Error bars represent standard error of the mean (SEM); n = 4. (B) Primary HMECs cultured in UL plates were infected with control (empty vector) CSCG lentivirus, or those encoding for PEA15-WT or PEA15-S116A mutant with similar infection efficiencies (as scored by GFP-positive cells). Two days following infection, 1 × 10 5 transduced cells were seeded in methylcellulose and total number of GFP-positive mammospheres formed was counted at the end of a week. Error bars represent standard error of the mean (SEM); n = 3. (C) Primary HMECs seeded in UL plates were treated with AMPK activator A769962 (100 μM) for 0 min, 30 min, 1 hr and 2 hrs, and subjected to immunoblot analysis for the specified proteins; n = 3. (D) Primary HMECs seeded in UL plates were subjected to immunoprecipitation with total AMPK antibody followed by immunoblotting for specified proteins. Rabbit immunoglobulin G (IgG)-treated lysate and resin alone served as controls, n = 3. (E) Commercially procured heterotrimeric AMPK and pure PEA15 were incubated in an in vitro kinase assay buffer in the presence of 50 μM adenosine triphosphate (ATP) and 0.1 mM AMP for 30 min, and then subjected to immunoblot analyses with commercially available antibodies that specifically recognize PEA15 Ser 116 phosphorylation; total PEA15 antibody served as a loading control. (F) Autoradiography of in vitro kinase assay using pure AMPK and PEA15 proteins performed in the presence of γ 32 P-labeled ATP (0.3 μCi) for 30 min; n = 3.

    Article Snippet: For radioactive in vitro kinase assay, 1 μg of purified PEA15 was incubated in the presence or absence of 1 μg of AMPK heterotrimer along with (γ32P) ATP (0.3 μCi) for 30 min at 30°C in AMPK kinase buffer (Cell Signaling Technology) supplemented with 50 μM ATP and 0.1 mM AMP.

    Techniques: Cell Culture, Transfection, Small Interfering RNA, Mass Spectrometry, Infection, Plasmid Preparation, Mutagenesis, Immunoprecipitation, Incubation, In Vitro, Kinase Assay, Autoradiography, Labeling

    CDK5 Directly Binds to and Phos-phorylates to Activate CREB1 in GSCs (A) CP681301 treatment of mice xenografted explant culture tumors induced a clear down-regulation of CREB1 phosphorylation. (B) Western blot data using these tumor tissues also show downregulation of CREB1 phosphorylation. (C) We then investigated the mechanism by which CDK5 regulates CREB1. We found that Cdk5 can directly bind with CREB1, and CP681301 reduces the binding of CDK5 to CREB1. (D) An in vitro kinase assay showed that CDK5 can phosphorylate CREB1 at serine 133, and CP681301 can suppress this phosphorylation in a dose-dependent manner. (E) Diagram showing our proposed model where CDK5 directly phosphorylates and activates CREB1 without the requirement of cAMP/PKA in GSCs.

    Journal: Cell reports

    Article Title: CDK5 Inhibition Resolves PKA/cAMP-Independent Activation of CREB1 Signaling in Glioma Stem Cells

    doi: 10.1016/j.celrep.2018.04.016

    Figure Lengend Snippet: CDK5 Directly Binds to and Phos-phorylates to Activate CREB1 in GSCs (A) CP681301 treatment of mice xenografted explant culture tumors induced a clear down-regulation of CREB1 phosphorylation. (B) Western blot data using these tumor tissues also show downregulation of CREB1 phosphorylation. (C) We then investigated the mechanism by which CDK5 regulates CREB1. We found that Cdk5 can directly bind with CREB1, and CP681301 reduces the binding of CDK5 to CREB1. (D) An in vitro kinase assay showed that CDK5 can phosphorylate CREB1 at serine 133, and CP681301 can suppress this phosphorylation in a dose-dependent manner. (E) Diagram showing our proposed model where CDK5 directly phosphorylates and activates CREB1 without the requirement of cAMP/PKA in GSCs.

    Article Snippet: The in vitro kinase assay buffer, ATP (Cell Signaling Tech), and 0, 0.5 μM, and 1 μM of CP681301 were added and mixed well.

    Techniques: Mouse Assay, Western Blot, Binding Assay, In Vitro, Kinase Assay

    Hipk phosphorylates Slimb to inhibit Arm ubiquitination without any effect on Arm phosphorylation. ( A ) In an in vitro kinase assay, Arm is sequentially phosphorylated in the presence of CK1 and Sgg (lane 2), as detected by a phospho-specific antibody against Arm (S44/S48). The presence of GST-Hipk has no effect on the phosphorylation of Arm by these kinases (lane 4). Phospho-Arm (S44/S48) is not detected in the absence of CK1 and Sgg (lanes 1 and 3). ( B ) In a pull-down assay, GST-Hipk forms a complex with Myc-Slimb extracted from Drosophila adults. The GST moiety alone does not bind Myc-Slimb. ( C ) In an in vitro kinase assay using radiolabeled ATP, immunoprecipitated Myc-Slimb is phosphorylated in the presence of GST-Hipk (lane 3) but not in its absence (lane 2). ( D ) In an in vitro ubiquitination assay, purified Arm is ubiquitinated in the presence of Slimb and other components of the ubiquitination machinery (lane 2) but not in the absence of Slimb (lane 1). Preincubation of GST-Hipk with Slimb in a kinase assay inhibits its ability to ubiquitinate Arm (lane 3). Preincubation of GST-Hipk with Slimb in the absence of ATP does not reduce Arm ubiquitination in the subsequent assay (lane 5). Preincubation of GST-Hipk with Arm in a kinase assay does not inhibit its Slimb-mediated ubiquitination (lane 6). ( E ) Protein lysates from Drosophila wing discs were assayed for levels of ubiquitinated Arm. Using omb-Gal4 to overexpress hipk results in lower levels of ubiquitinated Arm relative to wild-type discs. In the presence of the proteasome inhibitor MG132, wing discs reduced in function for hipk have higher levels of ubiquitinated Arm compared with wild-type discs. ( F ) In Drosophila S2 cells, Hipk enhances both the levels of cytosolic Arm (lane 2) and phospho-Arm (S44/S48) (lane 5), an effect that resembles the treatment of S2 cells with MG132 (lanes 3 and 6).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Drosophila homeodomain-interacting protein kinase inhibits the Skp1-Cul1-F-box E3 ligase complex to dually promote Wingless and Hedgehog signaling

    doi: 10.1073/pnas.1017548108

    Figure Lengend Snippet: Hipk phosphorylates Slimb to inhibit Arm ubiquitination without any effect on Arm phosphorylation. ( A ) In an in vitro kinase assay, Arm is sequentially phosphorylated in the presence of CK1 and Sgg (lane 2), as detected by a phospho-specific antibody against Arm (S44/S48). The presence of GST-Hipk has no effect on the phosphorylation of Arm by these kinases (lane 4). Phospho-Arm (S44/S48) is not detected in the absence of CK1 and Sgg (lanes 1 and 3). ( B ) In a pull-down assay, GST-Hipk forms a complex with Myc-Slimb extracted from Drosophila adults. The GST moiety alone does not bind Myc-Slimb. ( C ) In an in vitro kinase assay using radiolabeled ATP, immunoprecipitated Myc-Slimb is phosphorylated in the presence of GST-Hipk (lane 3) but not in its absence (lane 2). ( D ) In an in vitro ubiquitination assay, purified Arm is ubiquitinated in the presence of Slimb and other components of the ubiquitination machinery (lane 2) but not in the absence of Slimb (lane 1). Preincubation of GST-Hipk with Slimb in a kinase assay inhibits its ability to ubiquitinate Arm (lane 3). Preincubation of GST-Hipk with Slimb in the absence of ATP does not reduce Arm ubiquitination in the subsequent assay (lane 5). Preincubation of GST-Hipk with Arm in a kinase assay does not inhibit its Slimb-mediated ubiquitination (lane 6). ( E ) Protein lysates from Drosophila wing discs were assayed for levels of ubiquitinated Arm. Using omb-Gal4 to overexpress hipk results in lower levels of ubiquitinated Arm relative to wild-type discs. In the presence of the proteasome inhibitor MG132, wing discs reduced in function for hipk have higher levels of ubiquitinated Arm compared with wild-type discs. ( F ) In Drosophila S2 cells, Hipk enhances both the levels of cytosolic Arm (lane 2) and phospho-Arm (S44/S48) (lane 5), an effect that resembles the treatment of S2 cells with MG132 (lanes 3 and 6).

    Article Snippet: Kinase assays were performed by using kinase assay buffer/ATP (Cell Signaling Technology) with GST-Hipk and other proteins immunoprecipitated from Drosophila S2 cells.

    Techniques: In Vitro, Kinase Assay, Pull Down Assay, Immunoprecipitation, Ubiquitin Assay, Purification, Subsequent Assay