Rangap1

Manufactured by Abcam

Sourced in United Kingdom

RanGAP1 is a protein that plays a crucial role in the regulation of the Ran GTPase system. It functions as a GTPase-activating protein (GAP) for the Ran protein, stimulating the hydrolysis of Ran-bound GTP to GDP. This activity is essential for the maintenance of the Ran GTPase gradient, which is crucial for various cellular processes, including nucleocytoplasmic transport, spindle assembly, and cell cycle progression.

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Lab products found in correlation

Tubulin, by Merck Group (1 mentions) Ecl reagent, by Cytiva (1 mentions) Sumo2 3, by Cell Signaling Technology (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) Western lightning plus ecl detection reagent, by PerkinElmer (1 mentions) Sumo2 3, by Abcam (1 mentions) Flag m2, by Merck Group (1 mentions) Chemidoc xrs system, by Bio-Rad (1 mentions) Histone h3, by Abcam (1 mentions) Ha 3f10, by Merck Group (1 mentions) Myc 9e10, by Santa Cruz Biotechnology (1 mentions) Nf κb p65 d14e12, by Cell Signaling Technology (1 mentions) Anti trfp antibody, by Evrogen (1 mentions) Anti histone h3, by Abcam (1 mentions)

3 protocols using rangap1

Western Blotting for SUMO Pathway Proteins

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Whole cell lysates or tumor extracts were fractionated by non-reducing SDS–PAGE and immunoblotted with antibodies to SAE2 (for cell lysates: polyclonal rabbit antibody generated by Millennium, antibody was validated on cell lines overexpressing SAE2; for tumor extract: epitomics T0083, rabbit), Ubc9 (epitomics 2426–1, rabbit), SUMO1 (epitomics S2227, rabbit), SUMO2/3 (Cell Signaling Technologies 4971, rabbit), RanGAP1 (abcam ab2081, goat), p53 (Cell signaling Technologies 9282, rabbit), p21 (Santa Cruz sc-397, rabbit), cleaved caspase 3 (Cell Signaling Technologies 9661, rabbit), TopoIIα (Cell signaling Technologies 12286, rabbit). All primary antibodies were used 1:1000 dilution. Secondary HRP-labeled antibodies to goat IgG (Millipore, 1:2000 dilution) were used for RanGAP1 primary antibody and blots were developed with ECL reagent (Amersham). For other primary antibodies, the secondary antibody was Alexa-680-labelled antibody to rabbit/mouse IgG (Invitrogen, 1:5000 dilution) and the blots were imaged using the Li-Cor Odyssey Infrared Imaging system. Tubulin (Sigma) was blotted for a loading control.

He X., Riceberg J., Pulukuri S.M., Grossman S., Shinde V., Shah P., Brownell J.E., Dick L., Newcomb J, & Bence N. (2015). Characterization of the Loss of SUMO Pathway Function on Cancer Cells and Tumor Proliferation. PLoS ONE, 10(4), e0123882.

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Western Blot Analysis of Protein Samples

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Cell lysates and protein samples were mixed 1:1 with Laemmli sample buffer and reduced by boiling in 5% β-mercaptoethanol. Samples were separated on 8% polyacrylamide gels, transferred to PVDF membranes and membranes were probed with primary antibodies against: E4bp4 (C18; Santa Cruz Biotech), SUMO2/3 (AbCam), FLAG (M2; Sigma Aldrich), 6X-His (4D11; AbCam), α-Tubulin (DM1A; eBioscience), RanGAP1 (AbCam), Histone H3 (AbCam). Appropriate HRP-conjugated secondary antibodies (AbCam) were used with Western Lightning® Plus-ECL detection reagents (Perkin Elmer) to determine chemiluminescence. Images of exposed blots were digitally acquired using the ChemiDoc™ XRS+ system (Bio-Rad).

Kostrzewski T., Borg A.J., Meng Y., Filipovic I., Male V., Wack A., DiMaggio P.A, & Brady H.J. (2018). Multiple levels of control determine how E4bp4/Nfil3 regulates NK cell development. Journal of immunology (Baltimore, Md. : 1950), 200(4), 1370-1381.

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Comprehensive Immunofluorescence Protein Labeling

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TDP-43 (CAC-TIP-TD-P09, Cosmo bio Tokyo, Japan), myc (9E10, Santa Cruz Biotechnology, Dallas, TX, USA), HA (3F10, Sigma), NF-κB p65 (D14E12, Cell Signaling Technology, Danvers, MA, USA), Ran (ab53775, Abcam, Cambridge, UK), RanGAP1 (ab92360, Abcam), Anti-Histone H3 (ab10799, Abcam), LAMP1 (Ly1C6, Enzo life sciences, Farmingdale, NY, USA), TIAR (BD life sciences, Durham, NC, USA), Tumor Necrosis Factor alpha, human recombinant (rHuTNFα, 50435.50, Biomol, Hamburg, Germany), GAPDH (AM4300, ThermoFisher Scientific, Waltham, MA, USA). Anti-GFP (N86/8, UC Davis/NeuroMab Facility, UC Davis, CA, USA), Anti-tRFP antibody (AB233, Evrogen, Russian Federation).

Khosravi B., Hartmann H., May S., Möhl C., Ederle H., Michaelsen M., Schludi M.H., Dormann D, & Edbauer D. (2016). Cytoplasmic poly-GA aggregates impair nuclear import of TDP-43 in C9orf72 ALS/FTLD. Human Molecular Genetics, 26(4), 790-800.

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