Anti-SUMO2/3 Antibody, clone 3H12

Small ubiquitin-related modifier 2 (UniProt P61956; a.k.a. HSMT3, Sentrin-2, SMT3 homolog 2, Smt3B, SUMO-2, Ubiquitin-like protein SMT3B) and 3 (UniProt P55854; a.k.a. SMT3 homolog 1, Smt3A, SUMO-3, Ubiquitin-like protein SMT3A) are encoded by the SUMO2 (a.k.a. SMT3B, SMT3H2; Gene ID 6613) and SUMO3 (a.k.a. SMT3A, SMT3H1; Gene ID 6612) genes in human. SUMOylation, protein posttranslation modification by small ubiquitin-like modifier (SUMO), is a signaling event in many cellular processes. SUMO proteins are translated as immature precursors and subsequently converted to their mature forms through the activity of sentrin/SUMO-specific proteases (SENPs). As is the case with ubiquitination, SUMOylation is a reversible process. SUMO E1 activating enzyme, E2 conjugating enzyme, and E3 ligase mediate SUMOylation of substrate proteins, while SENPs are responsible for the deSUMOylation. SUMOylation usually occurs at lysine residues in the consensus KxD/E motif, although not all such lysines become SUMOylated and SUMOylation can also occur on lysine residues outside of this motif. SUMO2 and 3 share 97% identity at the amino acid level, while SUMO1 shares approximately 50% and SUMO4 shares about 87% identity with SUMO2/3. SUMO4 is the only SUMO member whose ctivation and conjugation has not been demonstrated. In addition to difference in their target substrates, SUMO2/3 can be SUMOylated and form chains, whereas SUMO1 cannot and may serve as chain terminator.

Clone 3H12 detects SUMO2/3 and SUMO2/3-modified (SUMOylated) proteins. Clone 3H12 reacts with SUMO2/3, but not SUMO1 (Uchimura, Y., et al. (2006). J. Biol. Chem. 281(32):23180-23190).

Predicted to react with a broad-range of mammalian species based on high sequence homology.

GST-tagged recombinant full-length human SUMO3 (Uchimura, Y., et al. (2006). J. Biol. Chem. 281(32):23180-23190).

Detect SUMO2/3 and SUMO2/3-modified (SUMOylated) proteins using this rat monoclonal Anti-SUMO2/3 Antibody, clone 3H12, Cat. No. MABS1139, validated for use in Immunocytochemistry and Western blotting.

Immunocytochemistry Analysis: Undiluted hybridoma culture supernatant from a representative lot detected SUMO2/3 immunoreactivity in 4% paraformaldehyde-fixed, 0.3% Triton X-100-permeabilized HeLa cells.

Immunocytochemistry Analysis: 10 µg/mL of purified clone 3H12 detected SUMO2/3 immunoreactivity in primary mouse neural progenitor cells (Courtesy of Taro Tachibana, Ph.D., Osaka City University, Japan).

Western Blotting Analysis: 1 µg/mL of purified clone 3H12 detected SUMOylated proteins in HeLa cell lysate (Courtesy of Taro Tachibana, Ph.D., Osaka City University, Japan).

Immunocytochemistry Analysis: A representative lot detected SUMO-2/3 nuclear immunoreactive foci co-localized with those of MCAF1 and SUMO1 and enriched in heterochromatin proteins by indirect immunofluorescent staining of exponentially growing C-33A cells fixed with 4% paraformaldehyde and permeabilized by 0.2% Triton X-100. Trasfecting C-33A cells with SUMO2/3 shRNA or Discosoma sp. red fluorescent protein (DsRed) fusion containg SUMO2/3-interacting sequence (MCAF1 a.a. 965-975) abolished nuclear staining by clone 3H12 (Uchimura, Y., et al. (2006). J. Biol. Chem. 281(32):23180-23190).

Research Category
Signaling

Evaluated by Western Blotting in HeLa cell lysate.

Western Blotting Analysis: A 1:500 dilution of this hybridoma culture supernatant detected SUMO2/3-modified (SUMOylated) proteins in 10 µg of HeLa cell lysate.

Variable depending on the degree of SUMOylation and the size(s) of the SUMOylated protein(s). 10.61/10.52 kDa (SUMO2/SUMO3; propeptide removed) calculated.

Unpurified.

Format: Unpurified

Rat IgG2a hybridoma culture supernatant with 0.05% sodium azide.

Stable for 1 year at -20°C from date of receipt.

Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Concentration: Please refer to lot specific datasheet.

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