Anti-CTCF (1:1000; Millipore, 07-729), anti-SMC1A (1:1000: Bethyl, A300-055A), anti-SMC3 (1:1,000; Abcam, ab9263), anti-RAD21 (1:1000; Abcam, ab992), anti-FLAG (1:1000; Sigma-Aldrich, F1804), anti-NUP153 (1:1000; Abcam, ab24700), anti-GAPDH (1:10,000; Sigma-Aldrich, G9545), anti-Histone H3 (1:10,000; Abcam, ab1791), and anti-α-TUBULIN (1:11,000; Santa Cruz, sc-5286) were used in western blot analysis. Note that anti-NUP153 (Abcam, ab24700) can also detect NUP62 and was used to detect NUP62 by western blot analysis. Anti-Rpb1 NTD (3 µl, Cell Signaling, 14958), Anti-CTCF (3 µl, Cell Signaling, 2899S), and anti-SMC3 (3 µg, Abcam, ab9263) were used in ChIP. Anti-LAMIN B1 (1:450; Abcam, ab16048), anti-V5 (1:400; Thermo Fisher Scientific, R960-25), anti-FLAG M2 (1:250; Sigma-Aldrich, F1804), anti-IgG(H+L)-Alexa555 (1:500; Thermo Fisher Scientific, A-21427), and anti-IgG(H+L)-Alexa488 (1:400; Thermo Fisher Scientific, A-11008 and A-32723) were used in immunofluorescence.
Anti nup153
Manufactured by Abcam
Sourced in United States
Anti-Nup153 is a lab equipment product that functions as an antibody. It is used for the detection and analysis of the Nup153 protein, which is a component of the nuclear pore complex in cells.
Automatically generated - may contain errors
Lab products found in correlation
Anti sc35, by Merck Group (2 mentions)
Anti α tubulin, by Santa Cruz Biotechnology (1 mentions)
Anti histone h3, by Abcam (1 mentions)
Anti gapdh, by Merck Group (1 mentions)
Anti v5, by Thermo Fisher Scientific (1 mentions)
Anti flag, by Merck Group (1 mentions)
Anti ctcf, by Merck Group (1 mentions)
Anti flag m2, by Merck Group (1 mentions)
Anti smc3, by Abcam (1 mentions)
Anti lamin b1, by Abcam (1 mentions)
Anti smc1a, by Fortis Life Sciences (1 mentions)
Anti rpb1 ntd, by Cell Signaling Technology (1 mentions)
Anti ctcf, by Cell Signaling Technology (1 mentions)
Anti rad21, by Abcam (1 mentions)
Anti nucleolin, by Abcam (1 mentions)
Anti nup62, by BD (1 mentions)
Anti pabp, by Cell Signaling Technology (1 mentions)
Anti hnrnp c1 c2, by Santa Cruz Biotechnology (1 mentions)
Anti sam68, by Santa Cruz Biotechnology (1 mentions)
Anti α β tubulin, by Cell Signaling Technology (1 mentions)
7 protocols using anti nup153
1
Comprehensive Protein Detection Protocol
2
Protein Detection via Western Analysis and IF
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The primary antibodies for the following proteins were used for Western analysis and immunofluorescence (IF): anti-Nup62 (BD Biosciences #610497, used at 1:2000 for Western), anti-Nup98 (Abcam #45584, used at 1:1000 for Western), anti-Nup153 (Abcam #96462, used at 1:1000 for Western), anti-eIF4G (BD Biosciences #610536, used at 1:1000 for Western), anti-PABP (Cell Signaling Technology #4992, used at 1:1000 for Western), anti-nucleolin (Abcam #22758, used at 1:3000 for Western), anti-hnRNP-C1/C2 (Santa Cruz #32308, used at 1:500 for IF), anti-SC35 (Sigma #4045, used at 1:500 for IF), anti-Sam68 (Santa Cruz #sc333, used at 1:500 for IF), and anti-α/β-tubulin (Cell Signaling Technology #2148, used at 1:1000 for Western). Antibodies to 3Cpro were kindly provided by S. Amineva (Madison, WI, USA; Amineva et al., 2004 (link)) and antibodies to dsRNA were kindly provided by S. Bowden (VIDRL, Melbourne, VIC, Australia).
3
Fluorescent Microspheres and Nup153 in HeLa Cells
4
Western Blot Analysis of Signaling Proteins
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The cell lysates were separated by 10% SDS-PAGE and then transferred to PVDF membranes (Millipore) using standard electroblotting procedures. The blots were then blocked and incubated overnight at 4 °C with anti-NUP153 (Abcam, Cambridge, MA, USA), anti-VEGF (Abcam), anti-basic fibroblast growth factor (Abcam), anti-hepatocyte growth factor (Abcam), and anti-tubulin (Millipore) primary antibodies. Immunolabeling was detected using an enhanced chemiluminescence kit (GE Healthcare, Pittsburgh, PA, USA) according to the manufacturer's instructions.
5
Comprehensive Analysis of Nuclear Components
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The following primary antibodies were used: mouse monoclonal antibodies against LB1 (Zymed, San Francisco, CA, USA); LB2 (Abcam, Cambridge, MA, USA); LA/C (Millipore, Billerica, MA, USA); rabbit polyclonal antiserum against LB1 (Abcam); LA/C (Santa Cruz Biotechnology, Santa Cruz, CA, USA), trimethyl histone H3 (Lys27) (Millipore); γ-tubulin, β-tubulin and β-actin (Sigma, St. Louis, MO, USA); and BU1/75 (ICR1) rat monoclonal anti-BrdU antibody (Abcam). To analyze the localization of nuclear components, the following primary antibodies were used: anti-Nup153 (Abcam), anti-nuclear pore complex (mab 414; Covance, Princeton, NJ, USA), anti-sc-35 (Sigma), anti-LAP2β (BD Biosciences, San Diego, CA, USA), and anti-fibrillarin (Cytoskeleton Denver, CO, USA) mouse monoclonal antibodies, as well as antiactivated RNA Polymerase II monoclonal IgM (Covance).
For the Western blot analysis, horseradish-peroxidase-conjugated secondary antibodies (Bio-Rad Laboratories, Hercules, CA, USA) were used for detection. For the immunofluorescence analyses, Alexa fluorophore-conjugated anti-mouse, anti-rabbit, or anti-rat antibodies from Invitrogen were used.
Unless otherwise specified, the general reagents and chemicals were from Sigma, and the reagents for the cell cultures were from Invitrogen.
For the Western blot analysis, horseradish-peroxidase-conjugated secondary antibodies (Bio-Rad Laboratories, Hercules, CA, USA) were used for detection. For the immunofluorescence analyses, Alexa fluorophore-conjugated anti-mouse, anti-rabbit, or anti-rat antibodies from Invitrogen were used.
Unless otherwise specified, the general reagents and chemicals were from Sigma, and the reagents for the cell cultures were from Invitrogen.
6
Antibody Sources for Nuclear Proteins
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Rabbit antibodies against Impα2 (RCH1) and Impβ were gifts from D. Görlich (Max-Planck-Institut für Biophysikalische Chemie, Göttingen, Germany), and rabbit antibodies against the N-terminal domain of LBR were provided by R.W. Wozniak (University of Alberta, Edmonton, Alberta, Canada). Commercial antibodies were mAB414 (mouse; BabCo), anti-Nup153 (ab24700, mouse; Abcam), anti-actin (mouse; Sigma-Aldrich), anti-HA (MMS-101P, mouse; Covance), and anti-Giantin (ab24586, rabbit; Abcam). Peptide-specific rabbit antibodies to Nup98 (peptide: CNRDSENLASPSEYPENGER), Nup96 (CSLHHPPDRTSDSTPDPQRV), Nup205 (TPSLSETVNRDGPRQDTQAC), and NDC1 (CQASAEHQKRLQQFLEFKE) have been described previously (Mansfeld et al., 2006 (link); Laurell et al., 2011 (link)).
7
Targeted Cellular Signaling Analysis
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Antibodies used were as follows: anti-Paxillin and anti-GM130 antibodies (BD Transduction Laboratories, New Jersey, USA), anti-Akap8 and anti-Nup153 (Abcam, Cambridge, UK), anti-FAK, anti-PDI, anti-p-Atf2 T71, anti-Atf2, anti-Brg1, anti-Rpb1, anti-Cdk8, anti-Cdk9, anti-Histone H4, anti-H3K4me2, anti-H3K4me3, anti-H3K27Ac, anti-Histone H3, anti-Lamin A/C and anti-GAPDH (Cell Signaling Technologies, Danvers, MA, USA), as well as anti-Ambra1 antibody (Millipore, Billerica, MA, USA). Anti-rabbit or mouse peroxidaseconjugated secondary antibodies were purchased from Cell Signaling Technologies. Cell culture FAK-deficient Squamous Cell Carcinoma (SCC) cell lines were generated as described previously (18) . SCCs were maintained in Glasgow MEM containing 10% FCS, 2 mM L-glutamine, nonessential amino acids, sodium pyruvate and MEM vitamins at 37°C, 5% CO 2 . SCC FAK-WT cells were maintained in 1 mg/ml hygromycin B. siRNA FAK-WT or FAK -/-SCC cells were transiently transfected using HiPerFect (Qiagen, Manchester, UK), according to manufacturer's protocol with a final concentration of 80 or 100 nM siRNA respectively (Supplementary Material and Methods, Table 1). Cells were analyzed at 48 h post transfection.
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