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hek293 gp  (TaKaRa)


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    TaKaRa hek293 gp
    ( A ) HeLa CDC14B-YFP and HeLa YFP -inducible cell lines were grown in heavy and light medium, respectively, to compare the phospho-proteome by mass spectrometry (MS). m / z , mass/charge ratio; IMAC, immobilized metal affinity chromatography. ( B ) Molecular network of the hypophosphorylated proteins (hits) from (A). The network was constructed in Cytoscape to extract functional information and shows hits (bold font) and their most related genes (regular font) identified using a guilt-by-association approach. Coexpression and shared protein domains are denoted by purple and yellow lines, respectively. The identified phosphorylation sites are summarized in table S1. ( C ) Gene Ontology (GO) analysis of hypophosphorylated proteins from (B). FDR, false discovery rate. ( D ) Sequence logos of hypophosphorylated p-sites from (A). ( E ) RPE1 WT and RPE1 CDC14B KO cells were treated with 3.3 μM NOC for 24 hours to accumulate prometaphase cells with high MeCP2 pS92 signal. The MeCP2 pS92 /MeCP2 ratio was densitometrically measured and is depicted in the graph on the right-hand side of the immunoblot. The asterisk indicates nonspecific protein bands that are detected by the anti-MeCP2 pS92 antibody. Green lines depict position of specific bands. The quantification was performed on three independent experiments. ( F ) RPE1 Tet ON cells expressing CDC14B-mSc or the catalytic inactive CDC14B C314S -mSc were treated and analyzed as described in (E). Asterisk and lines as in (E). The quantification was performed on three independent experiments. a.u., arbitrary units. ( G ) In vitro dephosphorylation of MeCP2 pS92 by purified CDC14B. <t>HEK293T</t> cells were transfected with MeCP2-FLAG . MeCP2-FLAG protein was immunoprecipitated using FLAG beads and incubated with recombinant CDC14B-GFP or catalytic inactive CDC14B C314S -GFP in vitro. The MeCP2 pS92 /MeCP2 ratio was densitometrically measured and is depicted in the graph next to the immunoblot. The quantification was performed on three independent experiments. IP, immunoprecipitation. (E to G) Results are means ± SD. Two-tailed unpaired t test. n.s. (not significant) P > 0.05, ** P < 0.01, and **** P < 0.0001.
    Hek293 Gp, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 175 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/hek293+gp/HEK+293+Tet-Off+Advanced+Cell+Line/pmc09858502-132-7-9
    Average 94 stars, based on 175 article reviews
    hek293 gp - by Bioz Stars, 2026-09
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    Images

    1) Product Images from "The HIPK2/CDC14B-MeCP2 axis enhances the spindle assembly checkpoint block by promoting cyclin B translation"

    Article Title: The HIPK2/CDC14B-MeCP2 axis enhances the spindle assembly checkpoint block by promoting cyclin B translation

    Journal: Science Advances

    doi: 10.1126/sciadv.add6982

    ( A ) HeLa CDC14B-YFP and HeLa YFP -inducible cell lines were grown in heavy and light medium, respectively, to compare the phospho-proteome by mass spectrometry (MS). m / z , mass/charge ratio; IMAC, immobilized metal affinity chromatography. ( B ) Molecular network of the hypophosphorylated proteins (hits) from (A). The network was constructed in Cytoscape to extract functional information and shows hits (bold font) and their most related genes (regular font) identified using a guilt-by-association approach. Coexpression and shared protein domains are denoted by purple and yellow lines, respectively. The identified phosphorylation sites are summarized in table S1. ( C ) Gene Ontology (GO) analysis of hypophosphorylated proteins from (B). FDR, false discovery rate. ( D ) Sequence logos of hypophosphorylated p-sites from (A). ( E ) RPE1 WT and RPE1 CDC14B KO cells were treated with 3.3 μM NOC for 24 hours to accumulate prometaphase cells with high MeCP2 pS92 signal. The MeCP2 pS92 /MeCP2 ratio was densitometrically measured and is depicted in the graph on the right-hand side of the immunoblot. The asterisk indicates nonspecific protein bands that are detected by the anti-MeCP2 pS92 antibody. Green lines depict position of specific bands. The quantification was performed on three independent experiments. ( F ) RPE1 Tet ON cells expressing CDC14B-mSc or the catalytic inactive CDC14B C314S -mSc were treated and analyzed as described in (E). Asterisk and lines as in (E). The quantification was performed on three independent experiments. a.u., arbitrary units. ( G ) In vitro dephosphorylation of MeCP2 pS92 by purified CDC14B. HEK293T cells were transfected with MeCP2-FLAG . MeCP2-FLAG protein was immunoprecipitated using FLAG beads and incubated with recombinant CDC14B-GFP or catalytic inactive CDC14B C314S -GFP in vitro. The MeCP2 pS92 /MeCP2 ratio was densitometrically measured and is depicted in the graph next to the immunoblot. The quantification was performed on three independent experiments. IP, immunoprecipitation. (E to G) Results are means ± SD. Two-tailed unpaired t test. n.s. (not significant) P > 0.05, ** P < 0.01, and **** P < 0.0001.
    Figure Legend Snippet: ( A ) HeLa CDC14B-YFP and HeLa YFP -inducible cell lines were grown in heavy and light medium, respectively, to compare the phospho-proteome by mass spectrometry (MS). m / z , mass/charge ratio; IMAC, immobilized metal affinity chromatography. ( B ) Molecular network of the hypophosphorylated proteins (hits) from (A). The network was constructed in Cytoscape to extract functional information and shows hits (bold font) and their most related genes (regular font) identified using a guilt-by-association approach. Coexpression and shared protein domains are denoted by purple and yellow lines, respectively. The identified phosphorylation sites are summarized in table S1. ( C ) Gene Ontology (GO) analysis of hypophosphorylated proteins from (B). FDR, false discovery rate. ( D ) Sequence logos of hypophosphorylated p-sites from (A). ( E ) RPE1 WT and RPE1 CDC14B KO cells were treated with 3.3 μM NOC for 24 hours to accumulate prometaphase cells with high MeCP2 pS92 signal. The MeCP2 pS92 /MeCP2 ratio was densitometrically measured and is depicted in the graph on the right-hand side of the immunoblot. The asterisk indicates nonspecific protein bands that are detected by the anti-MeCP2 pS92 antibody. Green lines depict position of specific bands. The quantification was performed on three independent experiments. ( F ) RPE1 Tet ON cells expressing CDC14B-mSc or the catalytic inactive CDC14B C314S -mSc were treated and analyzed as described in (E). Asterisk and lines as in (E). The quantification was performed on three independent experiments. a.u., arbitrary units. ( G ) In vitro dephosphorylation of MeCP2 pS92 by purified CDC14B. HEK293T cells were transfected with MeCP2-FLAG . MeCP2-FLAG protein was immunoprecipitated using FLAG beads and incubated with recombinant CDC14B-GFP or catalytic inactive CDC14B C314S -GFP in vitro. The MeCP2 pS92 /MeCP2 ratio was densitometrically measured and is depicted in the graph next to the immunoblot. The quantification was performed on three independent experiments. IP, immunoprecipitation. (E to G) Results are means ± SD. Two-tailed unpaired t test. n.s. (not significant) P > 0.05, ** P < 0.01, and **** P < 0.0001.

    Techniques Used: Mass Spectrometry, Affinity Chromatography, Construct, Functional Assay, Sequencing, Western Blot, Expressing, In Vitro, De-Phosphorylation Assay, Purification, Transfection, Immunoprecipitation, Incubation, Recombinant, Two Tailed Test

    Related Articles

    Cell Culture:

    Article Title: The HIPK2/CDC14B-MeCP2 axis enhances the spindle assembly checkpoint block by promoting cyclin B translation
    Article Snippet: .. RPE1, human embryonic kidney (HEK) 293T, and HEK293-GP (GP2-293, Clontech) cells were cultured in Gibco Dulbecco’s Modified Eagle’s medium (DMEM)/F-12 (Thermo Fisher Scientific) medium supplemented with 10% fetal bovine serum (FBS), 1% l -glutamine, and 1% penicillin/streptomycin. ..

    Modification:

    Article Title: The HIPK2/CDC14B-MeCP2 axis enhances the spindle assembly checkpoint block by promoting cyclin B translation
    Article Snippet: .. RPE1, human embryonic kidney (HEK) 293T, and HEK293-GP (GP2-293, Clontech) cells were cultured in Gibco Dulbecco’s Modified Eagle’s medium (DMEM)/F-12 (Thermo Fisher Scientific) medium supplemented with 10% fetal bovine serum (FBS), 1% l -glutamine, and 1% penicillin/streptomycin. ..



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    ( A ) HeLa CDC14B-YFP and HeLa YFP -inducible cell lines were grown in heavy and light medium, respectively, to compare the phospho-proteome by mass spectrometry (MS). m / z , mass/charge ratio; IMAC, immobilized metal affinity chromatography. ( B ) Molecular network of the hypophosphorylated proteins (hits) from (A). The network was constructed in Cytoscape to extract functional information and shows hits (bold font) and their most related genes (regular font) identified using a guilt-by-association approach. Coexpression and shared protein domains are denoted by purple and yellow lines, respectively. The identified phosphorylation sites are summarized in table S1. ( C ) Gene Ontology (GO) analysis of hypophosphorylated proteins from (B). FDR, false discovery rate. ( D ) Sequence logos of hypophosphorylated p-sites from (A). ( E ) RPE1 WT and RPE1 CDC14B KO cells were treated with 3.3 μM NOC for 24 hours to accumulate prometaphase cells with high MeCP2 pS92 signal. The MeCP2 pS92 /MeCP2 ratio was densitometrically measured and is depicted in the graph on the right-hand side of the immunoblot. The asterisk indicates nonspecific protein bands that are detected by the anti-MeCP2 pS92 antibody. Green lines depict position of specific bands. The quantification was performed on three independent experiments. ( F ) RPE1 Tet ON cells expressing CDC14B-mSc or the catalytic inactive CDC14B C314S -mSc were treated and analyzed as described in (E). Asterisk and lines as in (E). The quantification was performed on three independent experiments. a.u., arbitrary units. ( G ) In vitro dephosphorylation of MeCP2 pS92 by purified CDC14B. HEK293T cells were transfected with MeCP2-FLAG . MeCP2-FLAG protein was immunoprecipitated using FLAG beads and incubated with recombinant CDC14B-GFP or catalytic inactive CDC14B C314S -GFP in vitro. The MeCP2 pS92 /MeCP2 ratio was densitometrically measured and is depicted in the graph next to the immunoblot. The quantification was performed on three independent experiments. IP, immunoprecipitation. (E to G) Results are means ± SD. Two-tailed unpaired t test. n.s. (not significant) P > 0.05, ** P < 0.01, and **** P < 0.0001.

    Journal: Science Advances

    Article Title: The HIPK2/CDC14B-MeCP2 axis enhances the spindle assembly checkpoint block by promoting cyclin B translation

    doi: 10.1126/sciadv.add6982

    Figure Lengend Snippet: ( A ) HeLa CDC14B-YFP and HeLa YFP -inducible cell lines were grown in heavy and light medium, respectively, to compare the phospho-proteome by mass spectrometry (MS). m / z , mass/charge ratio; IMAC, immobilized metal affinity chromatography. ( B ) Molecular network of the hypophosphorylated proteins (hits) from (A). The network was constructed in Cytoscape to extract functional information and shows hits (bold font) and their most related genes (regular font) identified using a guilt-by-association approach. Coexpression and shared protein domains are denoted by purple and yellow lines, respectively. The identified phosphorylation sites are summarized in table S1. ( C ) Gene Ontology (GO) analysis of hypophosphorylated proteins from (B). FDR, false discovery rate. ( D ) Sequence logos of hypophosphorylated p-sites from (A). ( E ) RPE1 WT and RPE1 CDC14B KO cells were treated with 3.3 μM NOC for 24 hours to accumulate prometaphase cells with high MeCP2 pS92 signal. The MeCP2 pS92 /MeCP2 ratio was densitometrically measured and is depicted in the graph on the right-hand side of the immunoblot. The asterisk indicates nonspecific protein bands that are detected by the anti-MeCP2 pS92 antibody. Green lines depict position of specific bands. The quantification was performed on three independent experiments. ( F ) RPE1 Tet ON cells expressing CDC14B-mSc or the catalytic inactive CDC14B C314S -mSc were treated and analyzed as described in (E). Asterisk and lines as in (E). The quantification was performed on three independent experiments. a.u., arbitrary units. ( G ) In vitro dephosphorylation of MeCP2 pS92 by purified CDC14B. HEK293T cells were transfected with MeCP2-FLAG . MeCP2-FLAG protein was immunoprecipitated using FLAG beads and incubated with recombinant CDC14B-GFP or catalytic inactive CDC14B C314S -GFP in vitro. The MeCP2 pS92 /MeCP2 ratio was densitometrically measured and is depicted in the graph next to the immunoblot. The quantification was performed on three independent experiments. IP, immunoprecipitation. (E to G) Results are means ± SD. Two-tailed unpaired t test. n.s. (not significant) P > 0.05, ** P < 0.01, and **** P < 0.0001.

    Article Snippet: RPE1, human embryonic kidney (HEK) 293T, and HEK293-GP (GP2-293, Clontech) cells were cultured in Gibco Dulbecco’s Modified Eagle’s medium (DMEM)/F-12 (Thermo Fisher Scientific) medium supplemented with 10% fetal bovine serum (FBS), 1% l -glutamine, and 1% penicillin/streptomycin.

    Techniques: Mass Spectrometry, Affinity Chromatography, Construct, Functional Assay, Sequencing, Western Blot, Expressing, In Vitro, De-Phosphorylation Assay, Purification, Transfection, Immunoprecipitation, Incubation, Recombinant, Two Tailed Test