SUMO antibody

Antigen: SUMO

Clonality: Polyclonal

Host: Rabbit

Reactivity: Mouse (Murine), Human, Rat (Rattus)

Application: ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN107152

Additional Information

Immunogen: This purified antibody wasprepared from rabbit serum after repeatedimmunizations with recombinant humanSUMO protein.

Format: Liquid (sterile filtered)

Description: Covalent modification of cellularproteins by the ubiquitin-like modifier SUMO (small ubiquitin-likemodifier) regulates various cellular processes, such as nucleartransport, signal transduction, stress responses and cell cycleprogression. But, in contrast to ubiquination, sumoylation doesnot tag proteins for degradation by the 26S proteasome, butrather seems to enhance stability or modulate their subcellularcompartmentalization. Ubiquitin-like proteins fall into twoclasses: the first class, ubiquitin-like modifiers (UBLs) function asmodifiers in a manner analogous to that of ubiquitin. Examplesof UBLs are SUMO, Rub1 (also called Nedd8), Apg8 and Apg12.Proteins of the second class include parkin, RAD23 and DSK2,are designated ubiquitin-domain proteins (UDPs). Theseproteins contain domains that are related to ubiquitin but areotherwise unrelated to each other. In contrast to UBLs, UDPsare not conjugated to other proteins. Once covalently attached tocellular targets, SUMO regulates protein:protein and protein:DNAinteractions, as well as localization and stability of the targetprotein. Sumoylation occurs in most eukaryotic systems, andSUMO is highly conserved from yeast to humans. Whereinvertebrates have only a single SUMO gene termed SMT3,three members of the SUMO family have been identified in vertebrates: SUMO-1 and the close homologues SUMO-2 and SUMO-3. SUMO has been called SMT3 (yeast), sentrin, PIC1,GMP1 and UBL1. SUMO has been shown to bind and regulate mammalian SP-RINGs (such as Mdm2, PIAS and PML),RanGAP1, RanBP2, p53, p73, HIPK2, TEL, c-Jun, Fas, Daxx, TNFRI, Topo-I, Topo-II, WRN, Sp100, I kappa B-alpha , Androgen receptor (AR), GLUT1/4, Drosophila Ttk69, Dorsal, CaMK, yeast Septins, and viral CMV-IE1/2, EBV-BZLF1, HPV/BPV-E1.These bindings implicate SUMO in the stabilization of the target proteins and/or their localization to subcellular complexes.SUMO has an apparent molecular weight of ~12kDa and human SUMO-1 (a 101 amino acid polypeptide) shares 50%sequence identity with SUMO-2 and SUMO-3 and with yeast SMT3. SUMO and ubiquitin only show about 18% homology, butboth possess a common three-dimensional structure characterized by a tightly packed globular fold with beta -sheets wrapped around an alpha -helix.

Specificity: This product is an IgG fraction antibody purified from monospecific antiserum by a multi-step processwhich includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against thebuffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum.

Application Details

Application Notes: Recommended Dilutions: For immunoblotting a 1:2,000 dilution is recommended. An 11.6 kDa band corresponding tohuman SUMO is detected. Most human cell lysates can be used as a positive control without induction or stimulation. ForELISA a 1:4,000 to 1:20,000 dilution is recommended. Researchers should determine optimal titers for other applications. Note: This purified polyclonal antibody reacts with human SUMO by western blot and ELISA. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation. This antibody using the specifiedconditions may recognize other prominent intrinsic bands (UBLs or conjugates). Other intrinsic bands are readily detectable atlower dilutions.

Concentration: 5.0 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at 4° C prior to restoration. Restore with 0.1 ml of deionized water (or equivalent). Forextended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Centrifuge product ifnot completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undilutedliquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of restoration.

Research Area: Signaling, Proteases, DNA/RNA

Restrictions: For Research Use only

Images

Immunoblot of hSUMO fusion protein. Anti-SUMO antibody, generated by immunization with recombinant human SUMO, was tested by immunoblot against a SUMO-GFP fusion protein after cleavage by proteases. Dilution of the antibody between 1:1,000 and 1:5,000 showed strong reactivity specifically with the SUMO portion of the fusion protein (arrowhead). In this blot the antibody was used at a 1:2000 dilution incubated overnight at 4