peroxidase-igg fraction monoclonal Search Results


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  • 93
    Jackson Immuno peroxidase igg fraction monoclonal mouse anti biotin
    Peroxidase Igg Fraction Monoclonal Mouse Anti Biotin, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 93/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Jackson Immuno peroxidase igg fraction monoclonal mouse anti digoxin
    Peroxidase Igg Fraction Monoclonal Mouse Anti Digoxin, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 91/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peroxidase igg fraction monoclonal mouse anti digoxin/product/Jackson Immuno
    Average 91 stars, based on 15 article reviews
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    90
    Jackson Immuno peroxidase igg fraction monoclonal mouse anti fluorescein
    Neuronal co-localization of <t>FITC–IgG</t> and <t>LAMP2</t> . High magnification confocal microscope images of brain slice sections from a JNPL3 transgenic mouse. Brain slices were incubated with FITC–IgG from a high titer Tau 379–408[pSer 396, 404 ] immunized mouse (green) and after sectioning co-stained with an antibody to LAMP2 (red), which is a marker of late endosomes and lysosomes. The merged image indicates areas of co-localization (green/yellow) between FITC–IgG and late endosomes/lysosomes, mainly in perinuclear areas. The neuronal morphology is clearly delineated by the regions of staining.
    Peroxidase Igg Fraction Monoclonal Mouse Anti Fluorescein, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 90/100, based on 81 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peroxidase igg fraction monoclonal mouse anti fluorescein/product/Jackson Immuno
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    97
    Jackson Immuno peroxidase conjugated igg fraction monoclonal mouse anti rabbit igg
    Depletion of MAD2L2 leads to premature binding of CDH1 to the APC/C in prometaphase. (A) CDH1 interacts with the APC/C within 60 min after nocodazole release. <t>Immunoprecipitation</t> with anti-CDC27 from U2OS cells arrested in nocodazole and at 30, 60, and 90 min after release blotted for CDH1 to monitor the association of CDH1 with the APC/C. (B) Silencing of MAD2L2 leads to premature association of CDH1 with CDC27. (C) Complementation of the premature association of CDH1 with CDC27. The cells in this experiment all stably express hMAD2L2-myc. siMAD2L2 silences both the endogenous MAD2L2 and the transgenic MAD2L2-myc. The siMAD2L2 3′UTR only silences the endogenous MAD2L2, leaving the MAD2L2-myc expressed. See also Fig. 1 C . (D) Quantification of the premature association of CDH1 with CDC27 in MAD2L2-depleted cells. The amount of CDH1 immunoprecipitated with CDC27 was normalized to the <t>IgG</t> signal for each immunoprecipitation and the ratio of the amount of CDH1 pulled down in nocodazole (time = 0) and at 60 min after release was calculated. The constitutive association of CDH1 with CDC27 in prometaphase in MAD2L2-silenced cells results in an increase in this ratio, which approaches 1. “n” represents the number of independent experiments. Error bars = SEM; P-value calculated with unpaired, two-tailed t test assuming equal variance. The additional blots contributing to this analysis are shown in Fig. S1 D . (E) Depletion of CDH1 prevents the rapid mitotic exit seen in cells lacking MAD2L2. The percentage of cells in G1 was assessed 60 min after nocodazole release. “n” represents the number of independent experiments. Error bars = SEM; p, unpaired t test. The control siRNA and siMAD2L2 data are reproduced from Fig. 1 D for comparison. The effectiveness of the siRNA protocol is illustrated in the panel on the right. (F) Silencing of MAD2L2, but not REV3, can rescue the mitotic delay in CDC20-depleted cells. Quantification of the time taken for control (Control siRNA; solid black line), MAD2L2-depleted (siMAD2L2; solid red line), REV3-depleted (siREV3; solid blue line), CDC20-depleted (siCDC20, dashed dark gray line), CDC20- and MAD2L2-depleted (siCDC20 + siMAD2L2, dashed red line), and CDC20- and REV3-depleted (siCDC20 + siREV3, dashed blue line) U2OS cells expressing mCherry-H2B to complete mitosis, assessed by time-lapse video microscopy (1 frame every 5 min). The plot shows the cumulative percentage of cells that completed mitosis, measured from NEBD to cytokinesis. “n” represents the number of cells examined, collected from at least three independent experiments. P-value for siCDC20 and siCDC20 + siREV3 vs. Control siRNA, siMAD2L2, and siMAD2L2 + siCDC20
    Peroxidase Conjugated Igg Fraction Monoclonal Mouse Anti Rabbit Igg, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 97/100, based on 807 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peroxidase conjugated igg fraction monoclonal mouse anti rabbit igg/product/Jackson Immuno
    Average 97 stars, based on 807 article reviews
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    93
    Jackson Immuno peroxidase igg fraction monoclonal mouse anti goat igg light chain specific
    Depletion of MAD2L2 leads to premature binding of CDH1 to the APC/C in prometaphase. (A) CDH1 interacts with the APC/C within 60 min after nocodazole release. <t>Immunoprecipitation</t> with anti-CDC27 from U2OS cells arrested in nocodazole and at 30, 60, and 90 min after release blotted for CDH1 to monitor the association of CDH1 with the APC/C. (B) Silencing of MAD2L2 leads to premature association of CDH1 with CDC27. (C) Complementation of the premature association of CDH1 with CDC27. The cells in this experiment all stably express hMAD2L2-myc. siMAD2L2 silences both the endogenous MAD2L2 and the transgenic MAD2L2-myc. The siMAD2L2 3′UTR only silences the endogenous MAD2L2, leaving the MAD2L2-myc expressed. See also Fig. 1 C . (D) Quantification of the premature association of CDH1 with CDC27 in MAD2L2-depleted cells. The amount of CDH1 immunoprecipitated with CDC27 was normalized to the <t>IgG</t> signal for each immunoprecipitation and the ratio of the amount of CDH1 pulled down in nocodazole (time = 0) and at 60 min after release was calculated. The constitutive association of CDH1 with CDC27 in prometaphase in MAD2L2-silenced cells results in an increase in this ratio, which approaches 1. “n” represents the number of independent experiments. Error bars = SEM; P-value calculated with unpaired, two-tailed t test assuming equal variance. The additional blots contributing to this analysis are shown in Fig. S1 D . (E) Depletion of CDH1 prevents the rapid mitotic exit seen in cells lacking MAD2L2. The percentage of cells in G1 was assessed 60 min after nocodazole release. “n” represents the number of independent experiments. Error bars = SEM; p, unpaired t test. The control siRNA and siMAD2L2 data are reproduced from Fig. 1 D for comparison. The effectiveness of the siRNA protocol is illustrated in the panel on the right. (F) Silencing of MAD2L2, but not REV3, can rescue the mitotic delay in CDC20-depleted cells. Quantification of the time taken for control (Control siRNA; solid black line), MAD2L2-depleted (siMAD2L2; solid red line), REV3-depleted (siREV3; solid blue line), CDC20-depleted (siCDC20, dashed dark gray line), CDC20- and MAD2L2-depleted (siCDC20 + siMAD2L2, dashed red line), and CDC20- and REV3-depleted (siCDC20 + siREV3, dashed blue line) U2OS cells expressing mCherry-H2B to complete mitosis, assessed by time-lapse video microscopy (1 frame every 5 min). The plot shows the cumulative percentage of cells that completed mitosis, measured from NEBD to cytokinesis. “n” represents the number of cells examined, collected from at least three independent experiments. P-value for siCDC20 and siCDC20 + siREV3 vs. Control siRNA, siMAD2L2, and siMAD2L2 + siCDC20
    Peroxidase Igg Fraction Monoclonal Mouse Anti Goat Igg Light Chain Specific, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 93/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peroxidase igg fraction monoclonal mouse anti goat igg light chain specific/product/Jackson Immuno
    Average 93 stars, based on 37 article reviews
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    91
    Jackson Immuno peroxidase igg fraction monoclonal mouse anti sheep igg light chain specific
    Depletion of MAD2L2 leads to premature binding of CDH1 to the APC/C in prometaphase. (A) CDH1 interacts with the APC/C within 60 min after nocodazole release. <t>Immunoprecipitation</t> with anti-CDC27 from U2OS cells arrested in nocodazole and at 30, 60, and 90 min after release blotted for CDH1 to monitor the association of CDH1 with the APC/C. (B) Silencing of MAD2L2 leads to premature association of CDH1 with CDC27. (C) Complementation of the premature association of CDH1 with CDC27. The cells in this experiment all stably express hMAD2L2-myc. siMAD2L2 silences both the endogenous MAD2L2 and the transgenic MAD2L2-myc. The siMAD2L2 3′UTR only silences the endogenous MAD2L2, leaving the MAD2L2-myc expressed. See also Fig. 1 C . (D) Quantification of the premature association of CDH1 with CDC27 in MAD2L2-depleted cells. The amount of CDH1 immunoprecipitated with CDC27 was normalized to the <t>IgG</t> signal for each immunoprecipitation and the ratio of the amount of CDH1 pulled down in nocodazole (time = 0) and at 60 min after release was calculated. The constitutive association of CDH1 with CDC27 in prometaphase in MAD2L2-silenced cells results in an increase in this ratio, which approaches 1. “n” represents the number of independent experiments. Error bars = SEM; P-value calculated with unpaired, two-tailed t test assuming equal variance. The additional blots contributing to this analysis are shown in Fig. S1 D . (E) Depletion of CDH1 prevents the rapid mitotic exit seen in cells lacking MAD2L2. The percentage of cells in G1 was assessed 60 min after nocodazole release. “n” represents the number of independent experiments. Error bars = SEM; p, unpaired t test. The control siRNA and siMAD2L2 data are reproduced from Fig. 1 D for comparison. The effectiveness of the siRNA protocol is illustrated in the panel on the right. (F) Silencing of MAD2L2, but not REV3, can rescue the mitotic delay in CDC20-depleted cells. Quantification of the time taken for control (Control siRNA; solid black line), MAD2L2-depleted (siMAD2L2; solid red line), REV3-depleted (siREV3; solid blue line), CDC20-depleted (siCDC20, dashed dark gray line), CDC20- and MAD2L2-depleted (siCDC20 + siMAD2L2, dashed red line), and CDC20- and REV3-depleted (siCDC20 + siREV3, dashed blue line) U2OS cells expressing mCherry-H2B to complete mitosis, assessed by time-lapse video microscopy (1 frame every 5 min). The plot shows the cumulative percentage of cells that completed mitosis, measured from NEBD to cytokinesis. “n” represents the number of cells examined, collected from at least three independent experiments. P-value for siCDC20 and siCDC20 + siREV3 vs. Control siRNA, siMAD2L2, and siMAD2L2 + siCDC20
    Peroxidase Igg Fraction Monoclonal Mouse Anti Sheep Igg Light Chain Specific, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 91/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peroxidase igg fraction monoclonal mouse anti sheep igg light chain specific/product/Jackson Immuno
    Average 91 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    Neuronal co-localization of FITC–IgG and LAMP2 . High magnification confocal microscope images of brain slice sections from a JNPL3 transgenic mouse. Brain slices were incubated with FITC–IgG from a high titer Tau 379–408[pSer 396, 404 ] immunized mouse (green) and after sectioning co-stained with an antibody to LAMP2 (red), which is a marker of late endosomes and lysosomes. The merged image indicates areas of co-localization (green/yellow) between FITC–IgG and late endosomes/lysosomes, mainly in perinuclear areas. The neuronal morphology is clearly delineated by the regions of staining.

    Journal: Frontiers in Psychiatry

    Article Title: Mechanistic Studies of Antibody-Mediated Clearance of Tau Aggregates Using an ex vivo Brain Slice Model

    doi: 10.3389/fpsyt.2011.00059

    Figure Lengend Snippet: Neuronal co-localization of FITC–IgG and LAMP2 . High magnification confocal microscope images of brain slice sections from a JNPL3 transgenic mouse. Brain slices were incubated with FITC–IgG from a high titer Tau 379–408[pSer 396, 404 ] immunized mouse (green) and after sectioning co-stained with an antibody to LAMP2 (red), which is a marker of late endosomes and lysosomes. The merged image indicates areas of co-localization (green/yellow) between FITC–IgG and late endosomes/lysosomes, mainly in perinuclear areas. The neuronal morphology is clearly delineated by the regions of staining.

    Article Snippet: Membranes were probed with antibodies to LAMP2 (Abl93; Chen et al., , gift from Dr. P. Mathews), IgG–FITC (200-032-037 Jackson Immunoresearch), or total tau (DAKO A0024).

    Techniques: Microscopy, Slice Preparation, Transgenic Assay, Incubation, Staining, Marker

    FITC labeled IgG from a high titer mouse co-localizes with phosphorylated and pathological tau within the endosomal/lysosomal system . Confocal microscope images of brain slice sections from JNPL3 transgenic mice and WT (bottom panel) mice. Slices were incubated with FITC–IgG from a high titer Tau 379–408[pSer 396, 404 ] immunized mouse (green) and after sectioning co-stained with an antibody to LAMP2 (red), a marker of late endosomes/lysosomes, and an antibody to Rab5 (red), a marker of early endosomes and the nuclear stain Hoechst 33342. The merged images indicate areas of co-localization between FITC–IgG and endosomes and lysosomes (orange/yellow), mostly in perinuclear areas. Slices were also co-stained with an antibody to MC1 (red), which recognizes a disease related conformational tau epitope and CP13 (red) which recognizes tau pSer202. The merged images indicate areas of co-localization between FITC–IgG and pathological tau (yellow), mostly in perinuclear areas. Minimal staining was observed in the WT mouse. Scale bar = 10 μm.

    Journal: Frontiers in Psychiatry

    Article Title: Mechanistic Studies of Antibody-Mediated Clearance of Tau Aggregates Using an ex vivo Brain Slice Model

    doi: 10.3389/fpsyt.2011.00059

    Figure Lengend Snippet: FITC labeled IgG from a high titer mouse co-localizes with phosphorylated and pathological tau within the endosomal/lysosomal system . Confocal microscope images of brain slice sections from JNPL3 transgenic mice and WT (bottom panel) mice. Slices were incubated with FITC–IgG from a high titer Tau 379–408[pSer 396, 404 ] immunized mouse (green) and after sectioning co-stained with an antibody to LAMP2 (red), a marker of late endosomes/lysosomes, and an antibody to Rab5 (red), a marker of early endosomes and the nuclear stain Hoechst 33342. The merged images indicate areas of co-localization between FITC–IgG and endosomes and lysosomes (orange/yellow), mostly in perinuclear areas. Slices were also co-stained with an antibody to MC1 (red), which recognizes a disease related conformational tau epitope and CP13 (red) which recognizes tau pSer202. The merged images indicate areas of co-localization between FITC–IgG and pathological tau (yellow), mostly in perinuclear areas. Minimal staining was observed in the WT mouse. Scale bar = 10 μm.

    Article Snippet: Membranes were probed with antibodies to LAMP2 (Abl93; Chen et al., , gift from Dr. P. Mathews), IgG–FITC (200-032-037 Jackson Immunoresearch), or total tau (DAKO A0024).

    Techniques: Labeling, Microscopy, Slice Preparation, Transgenic Assay, Mouse Assay, Incubation, Staining, Marker

    FITC labeled IgG from a high titer mouse and tau are present in enriched lysosome fractions . Immunoblotting analysis of enriched lysosome fractions from two separate JNPL3 mice brain preparations is shown, A and B respectively. Brain slices (400 μm) were incubated with FITC–IgG from a high titer Tau 379–408[pSer 396, 404 ] immunized mouse for 2 h. Post treatment slices were processed to obtain an enriched lysosome fraction. Several fractions were obtained and the presence of lysosomes was confirmed in LAMP2 positive fractions A2, B1, B2, and B3. Mouse kidney lysate was used as a positive control. The kidney control is not a purified kidney preparation but a whole organ lysate, hence it has more background staining when compared to the enriched lysosome fractions from brain. Lysosome fractions were also immunoblotted for FITC–IgG. Fractions A2, B1, B2, and B3 were positive for FITC–IgG. To test if the lysosome fractions also contained tau, the FITC–IgG blot was reprobed with an antibody to total tau (DAKO A0024). Lysosome fractions A2, B1, B2, B3, and kidney lysate, all were positive for tau. Importantly, most of the FITC–IgG was detected in fractions with the highest levels of tau (A2 and B1). Overall, these findings are in accordance with the histological findings.

    Journal: Frontiers in Psychiatry

    Article Title: Mechanistic Studies of Antibody-Mediated Clearance of Tau Aggregates Using an ex vivo Brain Slice Model

    doi: 10.3389/fpsyt.2011.00059

    Figure Lengend Snippet: FITC labeled IgG from a high titer mouse and tau are present in enriched lysosome fractions . Immunoblotting analysis of enriched lysosome fractions from two separate JNPL3 mice brain preparations is shown, A and B respectively. Brain slices (400 μm) were incubated with FITC–IgG from a high titer Tau 379–408[pSer 396, 404 ] immunized mouse for 2 h. Post treatment slices were processed to obtain an enriched lysosome fraction. Several fractions were obtained and the presence of lysosomes was confirmed in LAMP2 positive fractions A2, B1, B2, and B3. Mouse kidney lysate was used as a positive control. The kidney control is not a purified kidney preparation but a whole organ lysate, hence it has more background staining when compared to the enriched lysosome fractions from brain. Lysosome fractions were also immunoblotted for FITC–IgG. Fractions A2, B1, B2, and B3 were positive for FITC–IgG. To test if the lysosome fractions also contained tau, the FITC–IgG blot was reprobed with an antibody to total tau (DAKO A0024). Lysosome fractions A2, B1, B2, B3, and kidney lysate, all were positive for tau. Importantly, most of the FITC–IgG was detected in fractions with the highest levels of tau (A2 and B1). Overall, these findings are in accordance with the histological findings.

    Article Snippet: Membranes were probed with antibodies to LAMP2 (Abl93; Chen et al., , gift from Dr. P. Mathews), IgG–FITC (200-032-037 Jackson Immunoresearch), or total tau (DAKO A0024).

    Techniques: Labeling, Mouse Assay, Incubation, Positive Control, Purification, Staining

    Depletion of MAD2L2 leads to premature binding of CDH1 to the APC/C in prometaphase. (A) CDH1 interacts with the APC/C within 60 min after nocodazole release. Immunoprecipitation with anti-CDC27 from U2OS cells arrested in nocodazole and at 30, 60, and 90 min after release blotted for CDH1 to monitor the association of CDH1 with the APC/C. (B) Silencing of MAD2L2 leads to premature association of CDH1 with CDC27. (C) Complementation of the premature association of CDH1 with CDC27. The cells in this experiment all stably express hMAD2L2-myc. siMAD2L2 silences both the endogenous MAD2L2 and the transgenic MAD2L2-myc. The siMAD2L2 3′UTR only silences the endogenous MAD2L2, leaving the MAD2L2-myc expressed. See also Fig. 1 C . (D) Quantification of the premature association of CDH1 with CDC27 in MAD2L2-depleted cells. The amount of CDH1 immunoprecipitated with CDC27 was normalized to the IgG signal for each immunoprecipitation and the ratio of the amount of CDH1 pulled down in nocodazole (time = 0) and at 60 min after release was calculated. The constitutive association of CDH1 with CDC27 in prometaphase in MAD2L2-silenced cells results in an increase in this ratio, which approaches 1. “n” represents the number of independent experiments. Error bars = SEM; P-value calculated with unpaired, two-tailed t test assuming equal variance. The additional blots contributing to this analysis are shown in Fig. S1 D . (E) Depletion of CDH1 prevents the rapid mitotic exit seen in cells lacking MAD2L2. The percentage of cells in G1 was assessed 60 min after nocodazole release. “n” represents the number of independent experiments. Error bars = SEM; p, unpaired t test. The control siRNA and siMAD2L2 data are reproduced from Fig. 1 D for comparison. The effectiveness of the siRNA protocol is illustrated in the panel on the right. (F) Silencing of MAD2L2, but not REV3, can rescue the mitotic delay in CDC20-depleted cells. Quantification of the time taken for control (Control siRNA; solid black line), MAD2L2-depleted (siMAD2L2; solid red line), REV3-depleted (siREV3; solid blue line), CDC20-depleted (siCDC20, dashed dark gray line), CDC20- and MAD2L2-depleted (siCDC20 + siMAD2L2, dashed red line), and CDC20- and REV3-depleted (siCDC20 + siREV3, dashed blue line) U2OS cells expressing mCherry-H2B to complete mitosis, assessed by time-lapse video microscopy (1 frame every 5 min). The plot shows the cumulative percentage of cells that completed mitosis, measured from NEBD to cytokinesis. “n” represents the number of cells examined, collected from at least three independent experiments. P-value for siCDC20 and siCDC20 + siREV3 vs. Control siRNA, siMAD2L2, and siMAD2L2 + siCDC20

    Journal: The Journal of Cell Biology

    Article Title: Sequestration of CDH1 by MAD2L2 prevents premature APC/C activation prior to anaphase onset

    doi: 10.1083/jcb.201302060

    Figure Lengend Snippet: Depletion of MAD2L2 leads to premature binding of CDH1 to the APC/C in prometaphase. (A) CDH1 interacts with the APC/C within 60 min after nocodazole release. Immunoprecipitation with anti-CDC27 from U2OS cells arrested in nocodazole and at 30, 60, and 90 min after release blotted for CDH1 to monitor the association of CDH1 with the APC/C. (B) Silencing of MAD2L2 leads to premature association of CDH1 with CDC27. (C) Complementation of the premature association of CDH1 with CDC27. The cells in this experiment all stably express hMAD2L2-myc. siMAD2L2 silences both the endogenous MAD2L2 and the transgenic MAD2L2-myc. The siMAD2L2 3′UTR only silences the endogenous MAD2L2, leaving the MAD2L2-myc expressed. See also Fig. 1 C . (D) Quantification of the premature association of CDH1 with CDC27 in MAD2L2-depleted cells. The amount of CDH1 immunoprecipitated with CDC27 was normalized to the IgG signal for each immunoprecipitation and the ratio of the amount of CDH1 pulled down in nocodazole (time = 0) and at 60 min after release was calculated. The constitutive association of CDH1 with CDC27 in prometaphase in MAD2L2-silenced cells results in an increase in this ratio, which approaches 1. “n” represents the number of independent experiments. Error bars = SEM; P-value calculated with unpaired, two-tailed t test assuming equal variance. The additional blots contributing to this analysis are shown in Fig. S1 D . (E) Depletion of CDH1 prevents the rapid mitotic exit seen in cells lacking MAD2L2. The percentage of cells in G1 was assessed 60 min after nocodazole release. “n” represents the number of independent experiments. Error bars = SEM; p, unpaired t test. The control siRNA and siMAD2L2 data are reproduced from Fig. 1 D for comparison. The effectiveness of the siRNA protocol is illustrated in the panel on the right. (F) Silencing of MAD2L2, but not REV3, can rescue the mitotic delay in CDC20-depleted cells. Quantification of the time taken for control (Control siRNA; solid black line), MAD2L2-depleted (siMAD2L2; solid red line), REV3-depleted (siREV3; solid blue line), CDC20-depleted (siCDC20, dashed dark gray line), CDC20- and MAD2L2-depleted (siCDC20 + siMAD2L2, dashed red line), and CDC20- and REV3-depleted (siCDC20 + siREV3, dashed blue line) U2OS cells expressing mCherry-H2B to complete mitosis, assessed by time-lapse video microscopy (1 frame every 5 min). The plot shows the cumulative percentage of cells that completed mitosis, measured from NEBD to cytokinesis. “n” represents the number of cells examined, collected from at least three independent experiments. P-value for siCDC20 and siCDC20 + siREV3 vs. Control siRNA, siMAD2L2, and siMAD2L2 + siCDC20

    Article Snippet: To avoid background signal form the IgG heavy chain in the immunoprecipitation experiments, anti–mouse (115-035-174; Jackson ImmunoResearch Laboratories, Inc.) and –rabbit (211-032-171; Jackson ImmunoResearch Laboratories, Inc.) IgG light chain–specific secondary antibodies were used at 1:10,000 dilution.

    Techniques: Binding Assay, Immunoprecipitation, Stable Transfection, Transgenic Assay, Two Tailed Test, Expressing, Microscopy

    CDK11 associates with TREX/THOC in 293 T cells A. A schematic representation of CDK11 and CycL2. CDK11 contains 782 residues and migrates with an apparent molecular mass of 110 kDa. CycL2 contains 520 residues with an apparent molecular mass of 58 kDa. White ovals depict regions rich in glutamic acids (pE), the kinase domain of CDK11, cyclin boxes and the arginine-serine (RS)-rich domain in CycL2. B. Dendrogram of the CDK11 proteome. Mass spectrometry of proteins that co-immunoprecipitated with CDK11 identified TREX/THOC (blue circles), EJC (brown circles) and various OTHER (yellow circles) proteins. C. Co-immunoprecipitations of f:CDK11-interacting proteins. Immunoprecipitation of FLAG epitope-tagged CDK11 protein (f:CDK11) was followed by western blotting with anti-FLAG, UAP56, THOC1, CycL2, RBM8A, SAP18 and SCAMP2 antibodies. Lanes contain the following: lane 1, input; lane 2, co-immunoprecipitated proteins; lane 3, IgG control. D. Co-immunoprecipitations between endogenous proteins. Immunoprecipitation of the endogenous CDK11 protein (CDK11) was followed by western blotting with anti-UAP56, THOC5, THOC1 and RBM8A antibodies (lanes 3–7). Lanes are as in panel C.

    Journal: Cell host & microbe

    Article Title: CDK11 in TREX/THOC regulates HIV mRNA 3’ end processing

    doi: 10.1016/j.chom.2015.10.012

    Figure Lengend Snippet: CDK11 associates with TREX/THOC in 293 T cells A. A schematic representation of CDK11 and CycL2. CDK11 contains 782 residues and migrates with an apparent molecular mass of 110 kDa. CycL2 contains 520 residues with an apparent molecular mass of 58 kDa. White ovals depict regions rich in glutamic acids (pE), the kinase domain of CDK11, cyclin boxes and the arginine-serine (RS)-rich domain in CycL2. B. Dendrogram of the CDK11 proteome. Mass spectrometry of proteins that co-immunoprecipitated with CDK11 identified TREX/THOC (blue circles), EJC (brown circles) and various OTHER (yellow circles) proteins. C. Co-immunoprecipitations of f:CDK11-interacting proteins. Immunoprecipitation of FLAG epitope-tagged CDK11 protein (f:CDK11) was followed by western blotting with anti-FLAG, UAP56, THOC1, CycL2, RBM8A, SAP18 and SCAMP2 antibodies. Lanes contain the following: lane 1, input; lane 2, co-immunoprecipitated proteins; lane 3, IgG control. D. Co-immunoprecipitations between endogenous proteins. Immunoprecipitation of the endogenous CDK11 protein (CDK11) was followed by western blotting with anti-UAP56, THOC5, THOC1 and RBM8A antibodies (lanes 3–7). Lanes are as in panel C.

    Article Snippet: Anti-CDK11, SAP18, SCAMP2, THOC1, THOC5, UAP56, THOC7 (Abcam, 19393, 31748, 154181, 487, 47955, 86070, 72295), anti-CDK11, CycL1, CycL2, IgG, eiF4A3, RBM8A (Bethyl, A300–310A, A302-058A, A301–677A, MI10–102, A302–981A, A301-033A), anti-THOC6, THOC1, CK2, RNAPII N20, PAP (Santa Cruz, 101177, 136426, 6479, 899, 32915), anti-Ser2P, Ser5P (Covance, H5, H14), anti-FLAG, HA, IgM, IgG (Sigma-Aldrich, F3165, F7425, H3663, H6908, A4540, A5420), peroxidase-conjugated monoclonal mouse anti-IgG (Jackson ImmunoResearch, 211-032–171, 115-035–174), anti-CstF77 (David Bentley).

    Techniques: Mass Spectrometry, Immunoprecipitation, FLAG-tag, Western Blot