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  • 90
    Millipore s protein agarose bead slurry
    S Protein Agarose Bead Slurry, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher magnetic beads
    Magnetic Beads, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Santa Cruz Biotechnology protein agarose bead slurry
    Protein Agarose Bead Slurry, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher protein g beads slurry
    Interaction of the different YopD mutants with YopB. A. YopB-YopD complexes secreted in bacterial supernatants of yopD , yopD /pYopD and yopD expressing the different yopD mutants were precipitated by 1 h incubation with YopD mab (clone 248:19) followed by incubation with Dynabeads <t>Protein</t> G (Invitrogen). After 3 washes, beads were resuspended in 2X Laemmli sample buffer, and boiled. The eluted material (IP) and an aliquot of the bacterial supernatants (SN) were resolved in SDS-PAGE. Western blot was performed using anti-YopD and anti-YopB Mabs, and anti-mouse IR680 or IR800 secondary antibodies. Bands corresponding to YopB and YopD are indicated by arrows. Also present in the IP samples are a band originated from the beads (marked with an asterix), and a weak band of unknown origin that migrates right below YopB. B. Signal intensities of immunoprecipitated YopB and YopD were calculated using Odyssey imaging system software (LI-COR Biosience). YopB-YopD interaction for each mutant was calculated as the ratio between immunoprecipitated YopB and YopD. Results were normalized to yopD /pYopD.
    Protein G Beads Slurry, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Interaction of the different YopD mutants with YopB. A. YopB-YopD complexes secreted in bacterial supernatants of yopD , yopD /pYopD and yopD expressing the different yopD mutants were precipitated by 1 h incubation with YopD mab (clone 248:19) followed by incubation with Dynabeads Protein G (Invitrogen). After 3 washes, beads were resuspended in 2X Laemmli sample buffer, and boiled. The eluted material (IP) and an aliquot of the bacterial supernatants (SN) were resolved in SDS-PAGE. Western blot was performed using anti-YopD and anti-YopB Mabs, and anti-mouse IR680 or IR800 secondary antibodies. Bands corresponding to YopB and YopD are indicated by arrows. Also present in the IP samples are a band originated from the beads (marked with an asterix), and a weak band of unknown origin that migrates right below YopB. B. Signal intensities of immunoprecipitated YopB and YopD were calculated using Odyssey imaging system software (LI-COR Biosience). YopB-YopD interaction for each mutant was calculated as the ratio between immunoprecipitated YopB and YopD. Results were normalized to yopD /pYopD.

    Journal: PLoS ONE

    Article Title: Random Mutagenesis Identifies a C-Terminal Region of YopD Important for Yersinia Type III Secretion Function

    doi: 10.1371/journal.pone.0120471

    Figure Lengend Snippet: Interaction of the different YopD mutants with YopB. A. YopB-YopD complexes secreted in bacterial supernatants of yopD , yopD /pYopD and yopD expressing the different yopD mutants were precipitated by 1 h incubation with YopD mab (clone 248:19) followed by incubation with Dynabeads Protein G (Invitrogen). After 3 washes, beads were resuspended in 2X Laemmli sample buffer, and boiled. The eluted material (IP) and an aliquot of the bacterial supernatants (SN) were resolved in SDS-PAGE. Western blot was performed using anti-YopD and anti-YopB Mabs, and anti-mouse IR680 or IR800 secondary antibodies. Bands corresponding to YopB and YopD are indicated by arrows. Also present in the IP samples are a band originated from the beads (marked with an asterix), and a weak band of unknown origin that migrates right below YopB. B. Signal intensities of immunoprecipitated YopB and YopD were calculated using Odyssey imaging system software (LI-COR Biosience). YopB-YopD interaction for each mutant was calculated as the ratio between immunoprecipitated YopB and YopD. Results were normalized to yopD /pYopD.

    Article Snippet: The pellet of a 20μl Protein G-beads slurry (Dynabeads Invitrogen) was added to the immunocomplexes an incubated for 10 min while rocking.

    Techniques: Expressing, Incubation, SDS Page, Western Blot, Immunoprecipitation, Imaging, Software, Mutagenesis