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densitometry computer software fluorchem 9800  (Alpha Innotech)
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(A) Real time RT-PCR of Bmi1 in U2OS lysates expressing the indicated proteins. (B) Left panel: overexpressed polyoma epitope-tagged Bmi1 (2PY-Bmi1) was infected with either SYT-SSX2 or pOZ backbone. Cell lysates were immunoblotted with Bmi1 antibody, then stripped and reprobed with an anti polyoma tag antibody. Immunoblotting for alpha-tubulin was used as a loading control. Right panel: cell lysates of pOZ and SYT-SSX2-infected cells were immunoblotted for either mouse monoclonal or rabbit polyclonal Bmi1 antibody. Immunoblotting for alpha-tubulin was used as a loading control. (C) <t>Densitometry</t> plotting of pulse-chase analysis of 2PY-Bmi1 in pOZ and SYT-SSX2-infected cells. 2PY-Bmi1-expressing cells infected with pOZ and SYT-SSX2 were labeled for 1 hr. with S 35 –labeled Methionine and Cysteine. Labeled cells were chased at the indicated timepoints and immunoprecipitated with a polyoma tag antibody. Immunoprecipitated 2PY-Bmi1 band intensities were quantitated by densitometry and plotted with pOZ and SYT-SSX2 experiments indicated. Pulse-chase experiment was successfully replicated (n = 2). (D) Immunoprecipitation studies demonstrating the specificity of the immunoprecipitated 2PY-Bmi1 band used in pulse-chase analysis. Negative controls using polyoma peptide blocking (lane 3), no antibody (lane 4) and naïve U2OS cells (lane 1) are indicated.
Densitometry Computer Software Fluorchem 9800, supplied by Alpha Innotech, 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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computer densitometry  (Alpha Innotech)
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(A) Real time RT-PCR of Bmi1 in U2OS lysates expressing the indicated proteins. (B) Left panel: overexpressed polyoma epitope-tagged Bmi1 (2PY-Bmi1) was infected with either SYT-SSX2 or pOZ backbone. Cell lysates were immunoblotted with Bmi1 antibody, then stripped and reprobed with an anti polyoma tag antibody. Immunoblotting for alpha-tubulin was used as a loading control. Right panel: cell lysates of pOZ and SYT-SSX2-infected cells were immunoblotted for either mouse monoclonal or rabbit polyclonal Bmi1 antibody. Immunoblotting for alpha-tubulin was used as a loading control. (C) <t>Densitometry</t> plotting of pulse-chase analysis of 2PY-Bmi1 in pOZ and SYT-SSX2-infected cells. 2PY-Bmi1-expressing cells infected with pOZ and SYT-SSX2 were labeled for 1 hr. with S 35 –labeled Methionine and Cysteine. Labeled cells were chased at the indicated timepoints and immunoprecipitated with a polyoma tag antibody. Immunoprecipitated 2PY-Bmi1 band intensities were quantitated by densitometry and plotted with pOZ and SYT-SSX2 experiments indicated. Pulse-chase experiment was successfully replicated (n = 2). (D) Immunoprecipitation studies demonstrating the specificity of the immunoprecipitated 2PY-Bmi1 band used in pulse-chase analysis. Negative controls using polyoma peptide blocking (lane 3), no antibody (lane 4) and naïve U2OS cells (lane 1) are indicated.
Computer Densitometry, supplied by Alpha Innotech, 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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Image Search Results


(A) Real time RT-PCR of Bmi1 in U2OS lysates expressing the indicated proteins. (B) Left panel: overexpressed polyoma epitope-tagged Bmi1 (2PY-Bmi1) was infected with either SYT-SSX2 or pOZ backbone. Cell lysates were immunoblotted with Bmi1 antibody, then stripped and reprobed with an anti polyoma tag antibody. Immunoblotting for alpha-tubulin was used as a loading control. Right panel: cell lysates of pOZ and SYT-SSX2-infected cells were immunoblotted for either mouse monoclonal or rabbit polyclonal Bmi1 antibody. Immunoblotting for alpha-tubulin was used as a loading control. (C) Densitometry plotting of pulse-chase analysis of 2PY-Bmi1 in pOZ and SYT-SSX2-infected cells. 2PY-Bmi1-expressing cells infected with pOZ and SYT-SSX2 were labeled for 1 hr. with S 35 –labeled Methionine and Cysteine. Labeled cells were chased at the indicated timepoints and immunoprecipitated with a polyoma tag antibody. Immunoprecipitated 2PY-Bmi1 band intensities were quantitated by densitometry and plotted with pOZ and SYT-SSX2 experiments indicated. Pulse-chase experiment was successfully replicated (n = 2). (D) Immunoprecipitation studies demonstrating the specificity of the immunoprecipitated 2PY-Bmi1 band used in pulse-chase analysis. Negative controls using polyoma peptide blocking (lane 3), no antibody (lane 4) and naïve U2OS cells (lane 1) are indicated.

Journal: PLoS ONE

Article Title: The Synovial Sarcoma-Associated SYT-SSX2 Oncogene Antagonizes the Polycomb Complex Protein Bmi1

doi: 10.1371/journal.pone.0005060

Figure Lengend Snippet: (A) Real time RT-PCR of Bmi1 in U2OS lysates expressing the indicated proteins. (B) Left panel: overexpressed polyoma epitope-tagged Bmi1 (2PY-Bmi1) was infected with either SYT-SSX2 or pOZ backbone. Cell lysates were immunoblotted with Bmi1 antibody, then stripped and reprobed with an anti polyoma tag antibody. Immunoblotting for alpha-tubulin was used as a loading control. Right panel: cell lysates of pOZ and SYT-SSX2-infected cells were immunoblotted for either mouse monoclonal or rabbit polyclonal Bmi1 antibody. Immunoblotting for alpha-tubulin was used as a loading control. (C) Densitometry plotting of pulse-chase analysis of 2PY-Bmi1 in pOZ and SYT-SSX2-infected cells. 2PY-Bmi1-expressing cells infected with pOZ and SYT-SSX2 were labeled for 1 hr. with S 35 –labeled Methionine and Cysteine. Labeled cells were chased at the indicated timepoints and immunoprecipitated with a polyoma tag antibody. Immunoprecipitated 2PY-Bmi1 band intensities were quantitated by densitometry and plotted with pOZ and SYT-SSX2 experiments indicated. Pulse-chase experiment was successfully replicated (n = 2). (D) Immunoprecipitation studies demonstrating the specificity of the immunoprecipitated 2PY-Bmi1 band used in pulse-chase analysis. Negative controls using polyoma peptide blocking (lane 3), no antibody (lane 4) and naïve U2OS cells (lane 1) are indicated.

Article Snippet: Immunoprecipitated 2PY-Bmi1 was quantitated using densitometry computer software (FluorChem 9800; Alpha Innotech, San Leandro, CA).

Techniques: Quantitative RT-PCR, Expressing, Infection, Western Blot, Control, Pulse Chase, Labeling, Immunoprecipitation, Blocking Assay