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Confirmation of protein-protein interactions identified by the screens. A, Fluorescent microscopy images of bait-prey protein pairs tagged with YFP fragments. HTC75 cells stably coexpressing YFPn-tagged TRF1 and YFPc alone (negative control), YFPc-tagged TIN2 (positive control), YFPc-MDH1, or YFPc-SET were visualized live under a fluorescence microscope. Hoechst33342 was added to visualize the nuclei. B, A flow chart of secondary coprecipitation screens that were used to confirm the identified protein-protein interactions. C, Examples of coprecipitation screens. Cell extracts from 293T cells coexpressing FLAG-tagged candidate proteins with GST alone or GST-tagged telomeric bait proteins were incubated with <t>GSH-agarose</t> beads. The proteins bound to <t>GSH</t> <t>beads</t> were then dot-blotted and probed with anti-FLAG antibodies.
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Confirmation of protein-protein interactions identified by the screens. A, Fluorescent microscopy images of bait-prey protein pairs tagged with YFP fragments. HTC75 cells stably coexpressing YFPn-tagged TRF1 and YFPc alone (negative control), YFPc-tagged TIN2 (positive control), YFPc-MDH1, or YFPc-SET were visualized live under a fluorescence microscope. Hoechst33342 was added to visualize the nuclei. B, A flow chart of secondary coprecipitation screens that were used to confirm the identified protein-protein interactions. C, Examples of coprecipitation screens. Cell extracts from 293T cells coexpressing FLAG-tagged candidate proteins with GST alone or GST-tagged telomeric bait proteins were incubated with <t>GSH-agarose</t> beads. The proteins bound to <t>GSH</t> <t>beads</t> were then dot-blotted and probed with anti-FLAG antibodies.
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Confirmation of protein-protein interactions identified by the screens. A, Fluorescent microscopy images of bait-prey protein pairs tagged with YFP fragments. HTC75 cells stably coexpressing YFPn-tagged TRF1 and YFPc alone (negative control), YFPc-tagged TIN2 (positive control), YFPc-MDH1, or YFPc-SET were visualized live under a fluorescence microscope. Hoechst33342 was added to visualize the nuclei. B, A flow chart of secondary coprecipitation screens that were used to confirm the identified protein-protein interactions. C, Examples of coprecipitation screens. Cell extracts from 293T cells coexpressing FLAG-tagged candidate proteins with GST alone or GST-tagged telomeric bait proteins were incubated with <t>GSH-agarose</t> beads. The proteins bound to <t>GSH</t> <t>beads</t> were then dot-blotted and probed with anti-FLAG antibodies.
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Confirmation of protein-protein interactions identified by the screens. A, Fluorescent microscopy images of bait-prey protein pairs tagged with YFP fragments. HTC75 cells stably coexpressing YFPn-tagged TRF1 and YFPc alone (negative control), YFPc-tagged TIN2 (positive control), YFPc-MDH1, or YFPc-SET were visualized live under a fluorescence microscope. Hoechst33342 was added to visualize the nuclei. B, A flow chart of secondary coprecipitation screens that were used to confirm the identified protein-protein interactions. C, Examples of coprecipitation screens. Cell extracts from 293T cells coexpressing FLAG-tagged candidate proteins with GST alone or GST-tagged telomeric bait proteins were incubated with <t>GSH-agarose</t> beads. The proteins bound to <t>GSH</t> <t>beads</t> were then dot-blotted and probed with anti-FLAG antibodies.
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Chem Impex International wang polystyrene resin
Confirmation of protein-protein interactions identified by the screens. A, Fluorescent microscopy images of bait-prey protein pairs tagged with YFP fragments. HTC75 cells stably coexpressing YFPn-tagged TRF1 and YFPc alone (negative control), YFPc-tagged TIN2 (positive control), YFPc-MDH1, or YFPc-SET were visualized live under a fluorescence microscope. Hoechst33342 was added to visualize the nuclei. B, A flow chart of secondary coprecipitation screens that were used to confirm the identified protein-protein interactions. C, Examples of coprecipitation screens. Cell extracts from 293T cells coexpressing FLAG-tagged candidate proteins with GST alone or GST-tagged telomeric bait proteins were incubated with <t>GSH-agarose</t> beads. The proteins bound to <t>GSH</t> <t>beads</t> were then dot-blotted and probed with anti-FLAG antibodies.
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Santa Cruz Biotechnology protein g sepharose
Confirmation of protein-protein interactions identified by the screens. A, Fluorescent microscopy images of bait-prey protein pairs tagged with YFP fragments. HTC75 cells stably coexpressing YFPn-tagged TRF1 and YFPc alone (negative control), YFPc-tagged TIN2 (positive control), YFPc-MDH1, or YFPc-SET were visualized live under a fluorescence microscope. Hoechst33342 was added to visualize the nuclei. B, A flow chart of secondary coprecipitation screens that were used to confirm the identified protein-protein interactions. C, Examples of coprecipitation screens. Cell extracts from 293T cells coexpressing FLAG-tagged candidate proteins with GST alone or GST-tagged telomeric bait proteins were incubated with <t>GSH-agarose</t> beads. The proteins bound to <t>GSH</t> <t>beads</t> were then dot-blotted and probed with anti-FLAG antibodies.
Protein G Sepharose, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Pharmacia LKB Biotechnology Inc gammabind-plus-sepharose beads
Confirmation of protein-protein interactions identified by the screens. A, Fluorescent microscopy images of bait-prey protein pairs tagged with YFP fragments. HTC75 cells stably coexpressing YFPn-tagged TRF1 and YFPc alone (negative control), YFPc-tagged TIN2 (positive control), YFPc-MDH1, or YFPc-SET were visualized live under a fluorescence microscope. Hoechst33342 was added to visualize the nuclei. B, A flow chart of secondary coprecipitation screens that were used to confirm the identified protein-protein interactions. C, Examples of coprecipitation screens. Cell extracts from 293T cells coexpressing FLAG-tagged candidate proteins with GST alone or GST-tagged telomeric bait proteins were incubated with <t>GSH-agarose</t> beads. The proteins bound to <t>GSH</t> <t>beads</t> were then dot-blotted and probed with anti-FLAG antibodies.
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Image Search Results


Confirmation of protein-protein interactions identified by the screens. A, Fluorescent microscopy images of bait-prey protein pairs tagged with YFP fragments. HTC75 cells stably coexpressing YFPn-tagged TRF1 and YFPc alone (negative control), YFPc-tagged TIN2 (positive control), YFPc-MDH1, or YFPc-SET were visualized live under a fluorescence microscope. Hoechst33342 was added to visualize the nuclei. B, A flow chart of secondary coprecipitation screens that were used to confirm the identified protein-protein interactions. C, Examples of coprecipitation screens. Cell extracts from 293T cells coexpressing FLAG-tagged candidate proteins with GST alone or GST-tagged telomeric bait proteins were incubated with GSH-agarose beads. The proteins bound to GSH beads were then dot-blotted and probed with anti-FLAG antibodies.

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Genome-wide YFP Fluorescence Complementation Screen Identifies New Regulators for Telomere Signaling in Human Cells *

doi: 10.1074/mcp.M110.001628

Figure Lengend Snippet: Confirmation of protein-protein interactions identified by the screens. A, Fluorescent microscopy images of bait-prey protein pairs tagged with YFP fragments. HTC75 cells stably coexpressing YFPn-tagged TRF1 and YFPc alone (negative control), YFPc-tagged TIN2 (positive control), YFPc-MDH1, or YFPc-SET were visualized live under a fluorescence microscope. Hoechst33342 was added to visualize the nuclei. B, A flow chart of secondary coprecipitation screens that were used to confirm the identified protein-protein interactions. C, Examples of coprecipitation screens. Cell extracts from 293T cells coexpressing FLAG-tagged candidate proteins with GST alone or GST-tagged telomeric bait proteins were incubated with GSH-agarose beads. The proteins bound to GSH beads were then dot-blotted and probed with anti-FLAG antibodies.

Article Snippet: The cleared lysate was then transferred into a 96-well binding plate (Purelink Clarificaiton plate, Invitrogen) preloaded with 100 μl/well of glutathione Sepharose 4B beads (10% slurry) (GE Healthcare Bio-Sciences AB), and incubated for 2 h at 4 °C with gentle agitation.

Techniques: Microscopy, Stable Transfection, Negative Control, Positive Control, Fluorescence, Incubation