Anti importin β1

Manufactured by Santa Cruz Biotechnology

Anti-importin β1 is a laboratory reagent that targets the importin β1 protein. Importin β1 is a key protein involved in the nuclear import process, facilitating the transport of molecules into the cell nucleus. The anti-importin β1 reagent can be used to study the role and function of importin β1 in various cellular processes.

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Lab products found in correlation

Anti gapdh, by Santa Cruz Biotechnology (2 mentions) Anti importin 7, by Abcam (2 mentions) Anti importin 8, by Abcam (2 mentions) Alexa 568, by Thermo Fisher Scientific (1 mentions) Ccg 1423, by Santa Cruz Biotechnology (1 mentions) Anti flag m2 affinity gel, by Merck Group (1 mentions) Horseradish peroxidase hrp conjugated donkey anti rabbit igg, by GE Healthcare (1 mentions) Docetaxel, by Selleck Chemicals (1 mentions) Nocodazole, by Merck Group (1 mentions) Paclitaxel, by Selleck Chemicals (1 mentions) Anti ar, by Merck Group (1 mentions) Cabazitaxel, by Selleck Chemicals (1 mentions) Anti β actin, by Santa Cruz Biotechnology (1 mentions) Anti histone h3, by Cell Signaling Technology (1 mentions) Anti gapdh, by Merck Group (1 mentions) Lncap, by American Type Culture Collection (1 mentions) Cos 7, by American Type Culture Collection (1 mentions)

4 protocols using anti importin β1

Antibodies and Reagents for Nrf2 Study

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CCG-1423, anti-NF-E2 related factor 2 (Nrf2), and anti-importin β1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Jasplakinolide (Jasp) and latrunculin B (LatB) were purchased ftom Caymen Chemical (Ann Arbor, MI). Other antibodies used in this study were anti-Flag M2 affinity gel (Sigma, St. Louis, MO), anti-HA affinity matrix and anti-HA (3F10) antibody (Roche Applied Science, Mannheim, Germany), and anti-DYKDDDDK (anti-Flag) antibody (Trans Genic, Kobe, Japan). Secondary antibodies and phalloidin were conjugated to Alexa 568 (Molecular Probes, Eugene, OR). We produced a polyclonal antibody against MRTF-A. In brief, a peptide consisting of amino acids 714–728 of mouse MRTF-A (MAL met; GenBank accession number: BC050941.1) was injected into a rabbit, and the antiserum was subjected to affinity purification.

Hayashi K., Watanabe B., Nakagawa Y., Minami S, & Morita T. (2014). RPEL Proteins Are the Molecular Targets for CCG-1423, an Inhibitor of Rho Signaling. PLoS ONE, 9(2), e89016.

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Immunoblotting Analysis of Importin Proteins in HeLa Cells

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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.

Roggero V.R., Zhang J., Parente L.E., Doshi Y., Dziedzic R.C., McGregor E.L., Varjabedian A.D., Schad S.E., Bondzi C, & Allison L.A. (2015). Nuclear Import of the Thyroid Hormone Receptor α1 is Mediated by Importin 7, Importin β1, and Adaptor Importin α1. Molecular and cellular endocrinology, 419, 185-197.

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Cell Line Characterization and Validation

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LNCaP, 22Rv1, and COS-7 cells were obtained from American Type Culture Collection. With the exception of drug-resistant lines, cells used in this study were within 20 passages (∼3 months of non-continuous culturing). All cell lines were tested and authenticated by the method of short tandem repeat profiling. Docetaxel, cabazitaxel, and paclitaxel were purchased from Selleck Chemicals (Houston, TX). Nocodazole was from Sigma Aldrich, and KX-01 was provided by Kinex Pharmaceuticals. The following antibodies were used in Western blot analysis: anti-GAPDH, anti-AR (N-terminus-directed, PG-21; Millipore), anti-importin β1, anti-β-actin (Santa Cruz), anti-p53 (Calbiochem), anti-histone H3 (Cell Signaling), and anti-AR-V7 (Precision Antibody).

Zhang G., Liu X., Li J., Ledet E., Alvarez X., Qi Y., Fu X., Sartor O., Dong Y, & Zhang H. (2015). Androgen receptor splice variants circumvent AR blockade by microtubule-targeting agents. Oncotarget, 6(27), 23358-23371.

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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.

Rosenberg D., Kolla L., Heo D., Cassio E., Veenstra M.J., Vakaki M.A., Zhang J., Anyetei‐Anum C.S., Allison L.A., & Buchser W. (2020). Modulating Intra-Nuclear LC3 with Small Molecules Rescues Cells from a Docetaxel-Induced Phenotype.

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