Anti sumo1

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-SUMO1 is a laboratory product that can be used to detect and study the Small Ubiquitin-like Modifier 1 (SUMO1) protein. SUMO1 is a post-translational modification involved in various cellular processes. The Anti-SUMO1 product provides a tool to investigate SUMO1 and its role in biological systems.

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Lab products found in correlation

Close spelling variants of this product

SUMO1 (20 citations) Rabbit anti-SUMO1 (5 citations) Anti-SUMO1 (C9H1, (2 citations)

16 protocols using anti sumo1

Protein Regulation and Circadian Rhythms

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Aβ, Dexamethasone (Dex), MG-132, Cycloheximide (CHX), and L-685,458 were purchased from the American Peptide Company (Sunnyvale, CA), Sigma-Aldrich (St. Louis, MO), and Calbiochem (La Jolla, CA). The following antibodies were used for immunodetection: anti-BMAL1, anti-CBP (Santa Cruz Biotechnology Inc., Santa Cruz, CA), anti-PER2 (Alpha Diagnostic International Inc., San Antonio, TX), anti-Sumo1 (Cell Signaling, Beverly, MA), anti-N-Cadherin (BD Biosciences, San Jose, CA), anti-Lamin A (Abcam, Cambridge, MA), and anti-α-Tubulin (Sigma-Aldrich, St Louis, MO).

Song H., Moon M., Choe H.K., Han D.H., Jang C., Kim A., Cho S., Kim K, & Mook-Jung I. (2015). Aβ-induced degradation of BMAL1 and CBP leads to circadian rhythm disruption in Alzheimer’s disease. Molecular Neurodegeneration, 10, 13.

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Lysis Conditions for Preserving Ubiquitin Conjugations

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Cells were washed with PBS and then lysed with either the lysis buffer (#9803; Cell Signaling Technology; 20 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1mM Na₂EDTA, 1 mM EGTA, 1% Triton, 2.5 mM sodium pyrophosphate, 1mM β-glycerophosphate, 1mM Na₃VO₄, 1 μg/mL leupeptin) or the acidic lysis buffer (50 mM HEPES, pH 6.0, 150 mM NaCl, 0.1% (w/v) SDS) supplemented with protease cocktail (Roche). The acidic lysis buffer was used to preserve the ubiquitin or ubiquitin-like conjugations (e.g., UBA3^N8, Ubc12^N8, UbcH7^Ub, Ubc9^SUMO) that are labile in the regular lysis buffer. The supernatant of the lysate was used for Western blotting. The antibodies used in this study include anti-cullin1 (H213; Santa cruz), anti-UBA3 (F-10; Santa cruz), anti-GlyRS (B01P; Abnova or sc-98614; Santa cruz), anti-SerRS (homemade), anti-V5 (R96-CUS; Invitrogen), and anti-Ube2F (pa5-26641; Thermo Fisher). Other antibodies including anti-NEDD8 (#2754), anti-Ubiquitin (#3936), anti-SUMO1 (#4930), anti-Ubc12 (#5641), anti-Ubc9 (#4918), anti-Flag (#2908), anti-UBA1(#4891), anti-UbcH7 (#3848), anti-UBA2 (#8688), anti-APPBP1 (#14321), anti-p27kip (#3698), and anti-α-Tubulin (#3873) are from Cell Signaling.

Mo Z., Zhang Q., Liu Z., Lauer J., Shi Y., Sun L., Griffin P.R, & Yang X.L. (2016). Neddylation requires glycyl-tRNA synthetase to protect activated E2. Nature structural & molecular biology, 23(8), 730-737.

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Analyzing Protein Expression in Cellular Compartments

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Cytoplasmic and nuclear fractions were separated by using a Qproteome cell compartment kit (Qiagen). Standard Western blot assays were used to analyze protein expression in cells. The following antibodies were used for assays: anti-p53 (1:2000 dilution; Santa Cruz Biotechnology, FL393), anti-Flag (1:40,000 dilution; Sigma, F7425), anti-Rac1 (1:4000 dilution; Millipore, 05-389), anti-HA (1:2000 dilution; Roche, 12013819001), anti-Myc (1:2000 dilution; Santa Cruz Biotechnology, SC-40 HRP), anti-SENP1 (1:2000 dilution; Cell Signaling Technology, 11929), anti-α-tubulin (1:2000 dilution; Santa Cruz Biotechnology, sc8035), anti-Histone H3 (1:1000 dilution; Cell Signaling Technology, 9715), anti-SUMO-1 (1:2000 dilution; Cell Signaling Technology, 4930), anti-p-PAK1/2 (1:1000 dilution; Cell Signaling Technology, 2606), anti-PAK1 (1:2000 dilution; Cell Signaling Technology, 2602), and anti-PAK2 (1:2000 dilution; Cell Signaling Technology, 2615).

Yue X., Zhang C., Zhao Y., Liu J., Lin A.W., Tan V.M., Drake J.M., Liu L., Boateng M.N., Li J., Feng Z, & Hu W. (2017). Gain-of-function mutant p53 activates small GTPase Rac1 through SUMOylation to promote tumor progression. Genes & Development, 31(16), 1641-1654.

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Lysis Conditions for Preserving Ubiquitin Conjugations

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Immunofluorescent Detection of SUMO-1 and α-Synuclein

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All samples were prepared using phosphate-buffered saline (PBS) supplemented with 10% goat serum and 0.1% Triton X-100. Next, the samples were incubated with blocking buffer (5% BSA in TBST, 1 TBS + 0.1% TWEEN 20 buffer) for 45 min at room temperature and then with anti-SUMO-1 (#4930, 1:100 dilution, Cell Signaling Technology, Boston, MA, USA) and anti-α-syn (#2628, 1:100 dilution, Cell Signaling Technology) primary antibodies, at 4°C overnight. After washed with PBS, the samples were incubated with secondary antibody [goat anti-rabbit IgG/fluorescein isothiocyanate (FITC) antibody (bs-0295G-FITC, 1:100 dilution, Bioss, Beijing, China)] for 1 h at room temperature. VECTASHIELD Antifade Mounting Medium with DAPI (H-1200, VECTOR, Burlingame, CA, USA) was used for nuclear labeling. Photomicrographs were captured using a fluorescence microscopy (A1+/A1R+, Nikon, Tokyo, Japan). All digital images were processed using the same settings to improve the contrast.

Zhu L.N., Qiao H.H., Chen L., Sun L.P., Hui J.L., Lian Y.L., Xie W.B., Ding J.Y., Meng Y.L., Zhu B.F, & Qiu P.M. (2018). SUMOylation of Alpha-Synuclein Influences on Alpha-Synuclein Aggregation Induced by Methamphetamine. Frontiers in Cellular Neuroscience, 12, 262.

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Western Blotting and Immunofluorescence Antibodies

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The following commercially available antibodies were used for Western blotting: mouse monoclonal antibodies against clathrin heavy chain (CHC; BD Biosciences; 610500), lamin A/C (Santa Cruz Biotechnology; sc-7292), Hsp90 (Santa Cruz Biotechnology; sc-13119), EHD2 (Santa Cruz Biotechnology; sc-100724), dynamin (BD Biosciences; 610245), and filamin A (Chemicon; MAB1678); rabbit polyclonal antibodies against SUMO2/3 (Cell Signaling Technology; 4971), Cav1 (BD Biosciences; 610059), pacsin2 (Abgent; AP8088b); and cavin1 (Sigma; AV36965); for immunofluorescence, mouse monoclonal anti-cavin1 (BD Biosciences; 611258), goat polyclonal anti-EHD2 (Abcam; Ab23935), rabbit polyclonal anti-Cav1 (BD Biosciences; 610059), mouse monoclonal anti-lamin A/C (Santa Cruz Biotechnology; sc-7292), rabbit polyclonal anti-SUMO1 (Cell Signaling Technology; 4930), and rabbit polyclonal anti-SUMO2/3 (Cell Signaling Technology; 4971). Antibodies conjugated to Alexa Fluor 488, Cy3, Cy5, or HRP (Beckman Coulter and Invitrogen) were used as secondary antibodies. HaloTag dye JF635 was provided by L. Lavis (Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA). Accutase was purchased from Sigma-Aldrich. Hepes, SDS, and Tris were purchased from Euromedex.

Torrino S., Shen W.W., Blouin C.M., Mani S.K., Viaris de Lesegno C., Bost P., Grassart A., Köster D., Valades-Cruz C.A., Chambon V., Johannes L., Pierobon P., Soumelis V., Coirault C., Vassilopoulos S, & Lamaze C. (2018). EHD2 is a mechanotransducer connecting caveolae dynamics with gene transcription. The Journal of Cell Biology, 217(12), 4092-4105.

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Quantitative Western Blot Analysis

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Western blot (WB) and immunoprecipitation (IP)/immunoblotting analyses were performed on whole cell lysates and nuclear fractions according to published methods [19 (link),20 (link)]. The anti-phospho-c-Abl (Tyr245), anti-β-catenin, anti-14-3-3 (pan), anti-phospho-14-3-3 (Ser186) and anti-Sumo 1 antibodies were purchased from Cell Signaling Technology. The anti-phospho-JNK (Thr 183) was purchased from Merck Millipore. The anti-CBY1 was previously described [16 (link)]. The anti-β-actin antibody used as loading control was purchased from Santa Cruz Biotechnology. The anti-histone H1 used as control for nuclear protein loading was purchased from Genetex. Signal intensities in single blots obtained in three separate experiments were measured by means of ChemiDoc-It instrument (UVP) equipped with a dedicated software (Launch VisionWorksLS from Euroclone). The differences among signal intensities were evaluated for statistical significance using the paired Student’s t-test.

Mancini M., Leo E., Takemaru K.I., Campi V., Castagnetti F., Soverini S., De Benedittis C., Rosti G., Cavo M., Santucci M.A, & Martinelli G. (2015). 14-3-3 Binding and Sumoylation Concur to the Down-Modulation of β-catenin Antagonist chibby 1 in Chronic Myeloid Leukemia. PLoS ONE, 10(7), e0131074.

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Comprehensive Immunofluorescence Antibody Panel

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The following primary antibodies were used:
Mouse Mab anti-mouse PML (mAb3739; Millipore), anti-human PML (clone 5E10; Roel van Driel or clone PG-M3; Santa Cruz), anti-SUMO-1 (clone 5B12; MBL), anti-SUMO-2/3 (clone 1E7; MBL), anti-NF160 (Invitrogen), anti-ßIII tubulin (MAB1637; Millipore), anti-ICP0 (Mab11060), anti-ICP4 (clone 10F1; Virusys), anti-ICP27 (Virusys); rabbit Mab anti-SUMO-1 (clone Y299; Abcam), anti-SUMO-2/3 (Mab4971; Cell Signaling): rabbit polyclonal anti-ATRX (H-300; Santa Cruz Biotechnology), anti-Daxx (M-112; Santa Cruz Biotechnology), anti-NF200 (Pierce), anti-SUMO-1 (4930; Cell Signaling), anti-SUMO-2/3 (ab3742; Abcam), anti-ICP0 (Rab190), anti-VP16 (ab4808; Abcam), and anti-pan-HSV-1 (LSBio) were used. All secondary antibodies were Alexa Fluor-conjugated and were raised in goats (Invitrogen).

Maroui M.A., Callé A., Cohen C., Streichenberger N., Texier P., Takissian J., Rousseau A., Poccardi N., Welsch J., Corpet A., Schaeffer L., Labetoulle M, & Lomonte P. (2016). Latency Entry of Herpes Simplex Virus 1 Is Determined by the Interaction of Its Genome with the Nuclear Environment. PLoS Pathogens, 12(9), e1005834.

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Western Blot Protein Analysis Protocol

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Cells were lysed in RIPA lysis buffer and the protein concentration of the cell lysates determined by BCA Protein Assay (Thermo Scientific, 23227). Equal amounts of protein were loaded onto SDS-PAGE gel and transferred to PVDF membranes. Western blotting was performed using primary antibodies and secondary antibodies conjugated with HRP. For immunoblotting, the following antibodies were used: anti-SENP2 (Abcam, ab58418, 1:1000), anti-HIF-1α (Cell Signaling Technology, 79233, 1:1000), anti-phospho-threonine (Cell Signaling Technology, 9386, 1:1000), anti-Ubc9 (Cell Signaling Technology, 4786, 1:1000), anti-BrdU (Cell Signaling Technology, 5292, 1:1000), anti-ubiquitin (Cell Signaling Technology, 3936, 1:1000), anti-cleaved-caspase 3 (Cell Signaling Technology, 9661, 1:1000), anti-SUMO1 (Cell Signaling Technology, 4930, 1:1000), anti-SUMO2/3 (Cell Signaling Technology, 4971, 1:1000), anti-SENP1 (Cell Signaling Technology, 11929, 1:1000), anti-SENP3 (Cell Signaling Technology, 5591, 1:1000), anti-HA-Tag (Cell Signaling Technology, 3724, 1:2000), anti-Flag-Tag (Cell Signaling Technology, 14793, 1:2000), anti-hexokinase 1 (Proteintech, 19662-1-AP, 1:1000), anti-hexokinase 2 (Proteintech, 22029-1-AP, 1:1000), anti-VDAC1 (Proteintech, 10866-1-AP, 1:1000), anti-alpha tubulin (Proteintech, 66031-1-Ig, 1:5000), anti-GAPDH (Proteintech, 10494-1-AP, 1:5000).

Shangguan X., He J., Ma Z., zhang W., Ji Y., Shen K., Yue Z., Li W., Xin Z., Zheng Q., Cao Y., Pan J., Dong B., Cheng J., Wang Q, & Xue W. (2021). SUMOylation controls the binding of hexokinase 2 to mitochondria and protects against prostate cancer tumorigenesis. Nature Communications, 12, 1812.

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Molecular Profiling of Protein Complexes

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Whole mouse brain or transfected cells were extracted and subsequently homogenized on ice in RIPA lysis buffer with protease inhibitors (TargetMol). Total protein levels were quantified using the BCA assay (Pierce). Equal protein amounts were separated by electrophoresis and transferred to PVDF membranes by electroblotting. Membranes were blocked with 5% nonfat dried milk, incubated overnight with primary antibodies, including anti-UBC9 and anti-SENP2 (Santa Cruz), anti-SUMO1 (Cell Signaling Technology), anti-SUMO2 and anti-CaM (Abcam), and anti-Kv7.2 and anti-Kv7.3 (Alomone Labs) antibodies. The membrane was washed and incubated with the secondary antibody coupled to peroxidase, and protein levels were detected with the chemo-luminescence system (Tanon). For immunoprecipitation experiments, the brain or cell lysate was incubated overnight with antibodies. All incubations were performed at 4 °C with constant agitation. Antibody-bound protein complexes were captured by the addition of protein A/G agarose and incubated for another 2 h. Protein A/G agarose was pelleted by centrifugation, and the immunoprecipitated protein complex was eluted using SDS-PAGE sample buffer and Western blotting with antibodies.

Chen X., Zhang Y., Ren X., Su Q., Liu Y., Dang X., Qin Y., Yang X., Xing Z., Shen Y., Wang Y., Bai Z., Yeh E.T., Wu H, & Qi Y. (2021). The SUMO-specific protease SENP2 plays an essential role in the regulation of Kv7.2 and Kv7.3 potassium channels. The Journal of Biological Chemistry, 297(4), 101183.

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