Twenty six hours post-transfection, HeLa cell protein lysates were prepared and analyzed by immunoblotting as described (Subramanian et al., 2015 (link)). Antibodies were used with the following concentrations: anti-GAPDH (Santa Cruz Biotechnology Inc, Dallas, TX), 1:5000; anti-importin β1 (Santa Cruz), 1:2000; anti-importin 4 (Santa Cruz), 1:333; anti-importin 5 (Santa Cruz), 1:10,000; anti-importin 7 (Abcam, Cambridge, MA), 1:1000; anti-importin 8 (Abcam), 1:250; anti-importin 9 (Abcam), 1:250; anti-importin 11 (Abcam), 1:333; anti-importin 13 (Santa Cruz), 1:100; anti-importin α1 (Santa Cruz), 1:2000; anti-importin α3 (Thermo Scientific), 0.2 μg/ml; horseradish peroxidase (HRP)-conjugated donkey anti-rabbit IgG (GE Healthcare Life Sciences), 1:25,000; HRP-sheep anti-mouse IgG (GE Healthcare Life Sciences), 1:25,000; or HRP-mouse anti-goat IgG (Santa Cruz Biotechnology), 1:25,000. Protein size was confirmed using Pre-Stained Kaleidoscope Protein Standards (Bio-Rad, Hercules, CA). X-ray films were quantified by scanning densitometry using NIH ImageJ software.
Anti importin 8
Manufactured by Abcam
Anti-importin 8 is a laboratory reagent that can be used to detect and measure the levels of importin 8 protein in biological samples. Importin 8 is a member of the importin protein family and plays a role in the transport of molecules into the cell nucleus. This product can be used in various research applications that involve the study of nuclear transport processes.
Automatically generated - may contain errors
2 protocols using anti importin 8
1
Immunoblotting Analysis of Importin Proteins in HeLa Cells
2
Importin Knockdown and Subcellular Localization
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Knockdown of importin β1, importin 7, and importin 8, compared to scrambled control shRNA, and protein compartment analysis was done by western blot as previously described 25 . Briefly, 786-0 cells in 6-well plates were transfected with sets of four shRNA plasmids (SABioscience, Frederick, MD) and lysed 24 hours post-transfection. Antibodies were used with the following concentrations: anti-GAPDH (Santa Cruz Biotechnology Inc, Dallas, TX), 1:5000; anti-importin β1 (Santa Cruz), 1:2000; anti-importin 7, 1:1000; anti-importin 8, 1:250 (both Abcam, Cambridge, MA). Independent replicates of each importin knockdown were completed from different cultures across different days.
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