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2 protocols using rabbit anti drosha

1

Western Blot Analysis of Protein Targets

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Western blotting of proteins was performed as described in published protocols (18 (link)). The cell lysates or immunoprecipitated proteins were analyzed by SDS-PAGE. After completion of electrophoresis, the proteins were transferred to a polyvinylidene fluoride or polyvinylidene difluoride (PVDF) membrane, followed by blocking at 4°C for a minimum of 1 h. The blots were probed overnight at 4°C with the primary antibodies. The antibodies used were as follows: mouse anti-Ago2 (Abnova), 1:1,000; rabbit anti-DICER1 (Bethyl), 1:8,000; rat anti-HA (Roche), 1:1,000; horseradish peroxidase (HRP)-conjugated anti-β-Actin (Sigma), 1:10000; rabbit anti-TRBP2 (Cell Signalling), 1:1000; rabbit anti-Drosha (Bethyl), 1:8,000; rabbit anti-P-p38 (Cell Signaling), 1:1,000; rabbit anti-P-ERK1/2 (Cell Signaling), 1:1,000; mouse anti-HSP70 (Santa Cruz Biotechnology), 1:1,000; rabbit anti-MSK1 (Cell Signaling), 1:1,000; mouse anti-HSP70 (Santa Cruz Biotechnology), 1:1,000; rabbit anti-cleaved PARP (Cell Signaling), 1:1,000; rabbit anti-cleaved caspase 9 (Cell Signaling), 1:1,000; and rabbit anti-P-Akt (Ser-473) (Cell Signaling), 1:1,000. Visualization of all Western blots was performed using an UVP BioImager 600 system equipped with VisionWorks Life Science software (UVP) V6.80. ImageJ software was used for densitometric analysis of blots for the relative quantification of bands.
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2

Western Blotting and Protein Analysis

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Western blotting of proteins has been followed as per published protocols (16081698). The cell lysates or immunoprecipitated proteins were electrophorated through SDS-PAGE. After completion of electrophoresis, proteins were transferred to polyvinylidene fluoride or polyvinylidene difluoride (PVDF) membrane followed by blocking at 4°C for minimum 1 h.
The blots were probed for overnight at 4°C with the following sets of primary antibodies: mouse anti-Ago2 (Abnova), 1:1000; rabbit anti-DICER1 (Bethyl) 1:8000; rat anti-HA (Roche), 1:1000; HRP-conjugated anti-β-Actin (SIGMA), 1:10000; rabbit anti-TRBP2 (Cell Signalling), 1:1000; rabbit anti-Drosha (Bethyl), 1:8000; rabbit P-p38 (Cell Signalling), 1:1000; rabbit anti-P-ERK1/2 (Cell Signalling), 1:1000; rabbit anti-P-MSK1 (Cell Signalling), 1:1000; rabbit anti-MSK1 (Cell Signalling), 1:1000; rabbit anti-HSP70 (Cell Signalling), 1:1000; rabbit anti-Cleaved PARP (Cell Signalling), 1:1000; rabbit anti-Cleaved Caspase 9 (Cell Signalling), 1:1000; rabbit anti-P-Akt (Ser-473) (Cell Signalling), 1:1000; Visualization of all Western blots was performed using an UVP BioImager 600 system equipped with VisionWorks Life Science software (UVP) V6.80. ImageJ software has been used for densitometric analysis of blots for relative quantification of bands.
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