mouse antibodies against sumo1 (Cell Signaling Technology Inc)

Structured Review

Mouse Antibodies Against Sumo1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse antibodies against sumo1/product/Cell Signaling Technology Inc
Average 95 stars, based on 1 article reviews
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1) Product Images from "SUMOylation of α-tubulin is a novel modification regulating microtubule dynamics"
Article Title: SUMOylation of α-tubulin is a novel modification regulating microtubule dynamics
Journal: Journal of Molecular Cell Biology
doi: 10.1093/jmcb/mjaa076

Figure Legend Snippet: α-tubulin is SUMOylated in cells and in vitro . ( A ) Immunoprecipitates with α-tubulin Ab from HEK293 cells expressing Flag-SUMO1, Flag-SUMO2, or Flag-SUMO3 were subject to immunoblotting (IB) and probed with Flag and α-tubulin (α-Tub) Abs. ( B ) Endogenous α-tubulin in HEK293 cells was immunoprecipitated and probed with SUMO1 Ab. ( C ) Immunoprecipitates with α-tubulin Ab from HEK293 cells expressing HA-Ubc9 were probed with HA and α-tubulin Abs. ( D ) Coomassie blue staining of purified mouse brain tubulin including α-tubulin and β-tubulin, indicated by arrows. ( E ) Coomassie blue staining of purified GST-SAE2/1, GST-Ubc9, His-SUMO1GG, and His-SUMO1ΔGG. Asterisk indicates the band of purified protein. ( F ) In vitro SUMOylation assay using purified GST-SAE2/1, GST-Ubc9, His-SUMO1GG, and brain tubulins. ( G ) Ratio of density of SUMOylated bands to unSUMOylated bands. ( H ) Purified tubulin was in vitro SUMOylated and probed with α-tubulin Ab. ( I ) In vitro SUMOylation assay using soluble tubulins and MTs. ( J ) Immunoprecipitates with β-tubulin Ab from HEK293 cells expressing Flag-SUMO1, Flag-SUMO2, or Flag-SUMO3 were probed with Flag and β-tubulin Abs. ( K ) Purified tubulin was in vitro SUMOylated and probed with β-tubulin Ab. ( L ) Endogenous β-tubulin in HEK293 cells was immunoprecipitated and probed with SUMO1 Ab. The experiments were repeated three times.
Techniques Used: In Vitro, Expressing, Western Blot, Immunoprecipitation, Staining, Purification

Figure Legend Snippet: SUMOylation is mainly enriched in soluble α-tubulin. ( A ) PLA with α-tubulin and SUMO1 Abs was performed in HEK293 cells. Confocal images of PLA signals and tubulin labelled after PLA are shown. The enlarged image of the boxed area is shown at the lower right. Scale bar, 10 μm. ( B ) PLA dots on and off MTs were quantified. n = 11 cells. ( C ) Soluble and polymerized tubulins were separated in SUMO1-overexpressing HEK293 cells and subject to IP using α-tubulin Ab. Immunoprecipitates were detected by SUMO1 Ab. The experiments were repeated three times. ( D ) Quantification of the SUMOylation in soluble or polymerized tubulins. Data are mean ± SEM from three independent experiments.
Techniques Used:

Figure Legend Snippet: α-tubulin is SUMOylated at K96, K166, and K304 and deSUMOylated by SENP1. ( A ) Alignment of lysines (in red color) and surrounding sequences of α-tubulin isotypes in the mouse. ( B ) Immunoprecipitates with Flag M2 beads from HEK293T cells expressing HA-SUMO1 with Flag-tagged isotypes of α-tubulin as indicated were probed with SUMO1 and Flag Abs. ( C ) Flag-tagged wild-type (WT) and various α1A mutant at MS-predicted SUMOylation sites were expressed with HA-SUMO1 in CHO-K1 cells and then immunoprecipitated. Immunoprecipitates were detected by SUMO1 and Flag Abs. ( D ) List of putative SUMOylation sites on α1A isotype predicted by different softwares. ( E ) Flag-tagged WT and combined mutant α1A at bioinformatics-predicted SUMOylation sites were expressed with HA-SUMO1 in CHO-K1 cells and then immunoprecipitated. Immunoprecipitates were detected by SUMO1 and Flag Abs. ( F ) Schematic representation of SUMOylation sites on the structure of α-tubulin. K96, K166, and K304 are shown as red spheres. ( G ) Location of SUMOylation sites and the interface for lateral contact between two α-tubulins in one MT. K96, K166, and K304 are shown as red spheres, and H2-S3 loop and M-loop are in blue. ( H ) Flag-α1A was overexpressed with HA-SUMO1 or HA-SUMO1-K7,16,17R mutant in HEK293 cells. Immunoprecipitates using Flag M2 beads were probed with HA and Flag Abs. ( I ) Immunoprecipitates with Flag M2 beads from HEK293 cells expressing Flag-SENP1 or Flag-SENP2 were probed with Flag and α-tubulin Abs. ( J ) Immunoprecipitates with α-tubulin from HEK293 cells expressing Flag-SENP1 or Flag-SENP2 with Flag-SUMO1 were probed with SUMO1 and α-tubulin Abs. ( K ) Immunoprecipitates from E13.5 brain of SENP1 +/+ and SENP1 –/– mice were probed with SUMO1 and α-tubulin Abs. The experiments were repeated three times.
Techniques Used: Expressing, Mutagenesis, Immunoprecipitation