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Envigo doxycycline (dox(+) condition)
Doxycycline (Dox(+) Condition), supplied by Envigo, 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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Average 90 stars, based on 1 article reviews
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Thermo Fisher dox condition culture medium
Selection of <t>dox-regulated</t> Tet-TNF-Cnx clones. Western blot analysis of Cnx in the selected overexpressing Tet-TNF-Cnx clones (2, 5, 6, 13, and 24) and a negative control cell (Tet-TNF-Null) to choose highly dox-regulatable clones. Cells were cultivated in the maintenance <t>medium</t> containing 1 μg/ml dox to regulate Cnx expression to a basal level. Two days after seeding, cells were cultivated for 24 h with 2 μg/ml dox or without dox after which they were sampled. Cell lysates were loaded at 5 × 104 cells/10 μl concentration. Relative band intensity was calculated using TINA 2.0 software. Regulation folds were calculated by dividing band intensity of 0 μg/ml dox <t>condition</t> by that of 2 μg/ml dox condition
Dox Condition Culture Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Selection of dox-regulated Tet-TNF-Cnx clones. Western blot analysis of Cnx in the selected overexpressing Tet-TNF-Cnx clones (2, 5, 6, 13, and 24) and a negative control cell (Tet-TNF-Null) to choose highly dox-regulatable clones. Cells were cultivated in the maintenance medium containing 1 μg/ml dox to regulate Cnx expression to a basal level. Two days after seeding, cells were cultivated for 24 h with 2 μg/ml dox or without dox after which they were sampled. Cell lysates were loaded at 5 × 104 cells/10 μl concentration. Relative band intensity was calculated using TINA 2.0 software. Regulation folds were calculated by dividing band intensity of 0 μg/ml dox condition by that of 2 μg/ml dox condition

Journal: Cell Stress & Chaperones

Article Title: Calnexin overexpression sensitizes recombinant CHO cells to apoptosis induced by sodium butyrate treatment

doi: 10.1007/s12192-008-0054-0

Figure Lengend Snippet: Selection of dox-regulated Tet-TNF-Cnx clones. Western blot analysis of Cnx in the selected overexpressing Tet-TNF-Cnx clones (2, 5, 6, 13, and 24) and a negative control cell (Tet-TNF-Null) to choose highly dox-regulatable clones. Cells were cultivated in the maintenance medium containing 1 μg/ml dox to regulate Cnx expression to a basal level. Two days after seeding, cells were cultivated for 24 h with 2 μg/ml dox or without dox after which they were sampled. Cell lysates were loaded at 5 × 104 cells/10 μl concentration. Relative band intensity was calculated using TINA 2.0 software. Regulation folds were calculated by dividing band intensity of 0 μg/ml dox condition by that of 2 μg/ml dox condition

Article Snippet: Regulation folds were calculated by dividing band intensity of 0 μg/ml dox condition by that of 2 μg/ml dox condition Culture medium and maintenance The medium for culture maintenance of Tet-TNF-Cnx and Tet-TNF-Null cells was IMDM supplemented with 10% dialyzed fetal bovine serum (Gibco, Grand Island, NY, USA), 250 μg/ml hygromycin, 350 μg/ml zeocin, 1 μg/ml dox (Clontech), and 0.02 μM methotrexate (MTX, Sigma).

Techniques: Selection, Clone Assay, Western Blot, Negative Control, Expressing, Concentration Assay, Software

Culture of Tet-TNF-Cnx clones with regulated Cnx expression. A Culture profile of Tet-TNF-Null (Null cells). B Culture profile of Tet-TNF-Cnx 2 (Clone 2, dox-non-responsive clone). C Culture profile of Tet-TNF-Cnx 5 (Clone 5 with low basal level Cnx). D Culture profile of Tet-TNF-Cnx 6 (clone 6 with high basal level Cnx). Spent medium was replaced with fresh medium containing control without dox (filled triangle), control with dox (empty triangle), 5 mM NaBu without dox (filled circle), and 5 mM NaBu with dox (empty circle) on day 2. Culture supernatant was collected everyday and the cells on the designated days for Western blot analysis. Cell number and viability percent was determined using the trypan blue dye exclusion method. Protein titer was quantitated by ELISA. Arrows indicate the time of NaBu addition. The error bars represent the standard deviation calculated from the data obtained in duplicate experiments

Journal: Cell Stress & Chaperones

Article Title: Calnexin overexpression sensitizes recombinant CHO cells to apoptosis induced by sodium butyrate treatment

doi: 10.1007/s12192-008-0054-0

Figure Lengend Snippet: Culture of Tet-TNF-Cnx clones with regulated Cnx expression. A Culture profile of Tet-TNF-Null (Null cells). B Culture profile of Tet-TNF-Cnx 2 (Clone 2, dox-non-responsive clone). C Culture profile of Tet-TNF-Cnx 5 (Clone 5 with low basal level Cnx). D Culture profile of Tet-TNF-Cnx 6 (clone 6 with high basal level Cnx). Spent medium was replaced with fresh medium containing control without dox (filled triangle), control with dox (empty triangle), 5 mM NaBu without dox (filled circle), and 5 mM NaBu with dox (empty circle) on day 2. Culture supernatant was collected everyday and the cells on the designated days for Western blot analysis. Cell number and viability percent was determined using the trypan blue dye exclusion method. Protein titer was quantitated by ELISA. Arrows indicate the time of NaBu addition. The error bars represent the standard deviation calculated from the data obtained in duplicate experiments

Article Snippet: Regulation folds were calculated by dividing band intensity of 0 μg/ml dox condition by that of 2 μg/ml dox condition Culture medium and maintenance The medium for culture maintenance of Tet-TNF-Cnx and Tet-TNF-Null cells was IMDM supplemented with 10% dialyzed fetal bovine serum (Gibco, Grand Island, NY, USA), 250 μg/ml hygromycin, 350 μg/ml zeocin, 1 μg/ml dox (Clontech), and 0.02 μM methotrexate (MTX, Sigma).

Techniques: Clone Assay, Expressing, Control, Western Blot, Enzyme-linked Immunosorbent Assay, Standard Deviation

Western blot analysis of Tet-TNF-Cnx clones for cellular proteins from batch culture with NaBu treatment. A Western blot data for Tet-TNF-Null (null cells). B Western blot data for Tet-TNF-Cnx 2 (clone 2, dox-non-responsive clone). C Western blot data for Tet-TNF-Cnx 5 (Clone 5 with low basal level Cnx). D Western blot data for Tet-TNF-Cnx 6 (Clone 6 with high basal level Cnx). Cells were loaded at a concentration of 1 × 105 cells/10 μl and resolved using the 4–20% Tris-glycine gel. Note the elevated cytochrome C release and PARP cleavage for clones 5 and 6 under Cnx-overexpressed condition (− dox)

Journal: Cell Stress & Chaperones

Article Title: Calnexin overexpression sensitizes recombinant CHO cells to apoptosis induced by sodium butyrate treatment

doi: 10.1007/s12192-008-0054-0

Figure Lengend Snippet: Western blot analysis of Tet-TNF-Cnx clones for cellular proteins from batch culture with NaBu treatment. A Western blot data for Tet-TNF-Null (null cells). B Western blot data for Tet-TNF-Cnx 2 (clone 2, dox-non-responsive clone). C Western blot data for Tet-TNF-Cnx 5 (Clone 5 with low basal level Cnx). D Western blot data for Tet-TNF-Cnx 6 (Clone 6 with high basal level Cnx). Cells were loaded at a concentration of 1 × 105 cells/10 μl and resolved using the 4–20% Tris-glycine gel. Note the elevated cytochrome C release and PARP cleavage for clones 5 and 6 under Cnx-overexpressed condition (− dox)

Article Snippet: Regulation folds were calculated by dividing band intensity of 0 μg/ml dox condition by that of 2 μg/ml dox condition Culture medium and maintenance The medium for culture maintenance of Tet-TNF-Cnx and Tet-TNF-Null cells was IMDM supplemented with 10% dialyzed fetal bovine serum (Gibco, Grand Island, NY, USA), 250 μg/ml hygromycin, 350 μg/ml zeocin, 1 μg/ml dox (Clontech), and 0.02 μM methotrexate (MTX, Sigma).

Techniques: Western Blot, Clone Assay, Concentration Assay