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Primary Antibodies  >  Ubiquitin and UBL Antibodies
 

SUMO Antibody

Rabbit Polyclonal
NCI Collaboration
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Western blot of SUMO-GFP fusion proteins cleaved by insect cell protein extracts.
Anti-SUMO antibody, generated by immunization with recombinant yeast SUMO, was tested by western blot against several constructs of SUMO-GFP fusion proteins after cleavage by proteases in insect cell protein extracts.  These constructs contained various linkers between the SUMO and GFP portion of the fusion proteins.  Each sample was run twice.  The left lanes each contain 2 ug E.coli expressed and purified SUMO-GFP fusion proteins after incubation with lysed cells (50 ug total protein) for 1 h.  The right lanes contain the same fusion proteins incubated with the lysate in the presence of 2% SDS.  After probing with anti-GFP antibodies the membranes were stripped of antibody using SDS-DTT solution for 30 m at 60° C and were then re-probed using the anti-SUMO antibody at a 1:1000 dilution incubated overnight at 4° C in 5% non-fat dry milk in TTBS.   Detection occurred using a 1:2000 dilution of HRP-labeled Donkey anti-Rabbit IgG (code # 611-703-127) for 1 hour at room temperature.  A chemiluminescence system was used for signal detection (Roche).  Other detection systems will yield similar results.
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Number:
200-401-428
Size:
500 µg
Price:
$309.00
Shipping:
$40.00
Handling:
$20.00
Availability:
Ships next business day.

SUMO Antibody Properties

Description
Anti-SUMO (Yeast) (RABBIT) Antibody - 200-401-428
Host
Rabbit
Target Species
Yeast
Known Cross Reactivity
yeast
Clonality
Polyclonal
Application
ELISA : 1:5,000 - 1:25,000
Western Blot : 1:500 - 1:3,000
Other Dilution: User Optimized
Physical State
Lyophilized
Shipping Condition
Ambient
Format
IgG
Label
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Reconstitution Volume
500 µL
Reconstitution Buffer
Restore with deionized water (or equivalent)
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
SMT3
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SUMO Antibody Description

Background
Anti SUMO Antibody recognizes SUMO. Covalent modification of cellular proteins by the ubiquitin-like modifier SUMO (small ubiquitin-like modifier) regulates various cellular processes, such as nuclear transport, signal transduction, stress responses and cell cycle progression.  But, in contrast to ubiquination, sumoylation does not tag proteins for degradation by the 26S proteasome, but rather seems to enhance stability or modulate their subcellular compartmentalization.  Ubiquitin-like proteins fall into two classes: the first class, ubiquitin-like modifiers (UBLs) function as modifiers in a manner analogous to that of ubiquitin.  Examples of 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).  These proteins contain domains that are related to ubiquitin but are otherwise unrelated to each other.  In contrast to UBLs, UDPs are not conjugated to other proteins.  Once covalently attached to cellular targets, SUMO regulates protein:protein and protein:DNA interactions, as well as localization and stability of the target protein.  Sumoylation occurs in most eukaryotic systems, and SUMO is highly conserved from yeast to humans.   Where invertebrates 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, IkB-a, 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, but both possess a common three-dimensional structure characterized by a tightly packed globular fold with b-sheets wrapped around an a-helix.
Synonyms
Ubiquitin-like protein SMT3 antibody, SMT3 antibody
Immunogen
This purified antibody was prepared from rabbit serum after repeated immunizations with recombinant yeast SUMO protein.
Immunogen Type
Recombinant Protein
Storage Condition
Store vial at 4° C prior to restoration.   For extended storage aliquot contents and freeze at -20° C or below.  Avoid cycles of freezing and thawing.  Centrifuge product if not completely clear after standing at room temperature.  This product is stable for several weeks at 4° C as an undiluted liquid.  Dilute only prior to immediate use. 
Application Note
This purified polyclonal antibody reacts yeast SUMO by western blot and ELISA. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation. For immunoblotting a 1:1,000 dilution is recommended.  A 12 kDa band corresponding to yeast SUMO is detected.  Most yeast cell lysates can be used as a positive control without induction or stimulation.  For ELISA a 1:1,000 to 1:5,000 dilution is recommended.   Researchers should determine optimal titers for other applications. For immunoblotting a 1:1,000 dilution is recommended.  A 12 kDa band corresponding to yeast SUMO is detected.  Most yeast cell lysates can be used as a positive control without induction or stimulation.  For ELISA a 1:1,000 to 1:5,000 dilution is recommended.   Researchers should determine optimal titers for other applications. For immunoblotting a 1:1,000 dilution is recommended.  A 12 kDa band corresponding to yeast SUMO is detected.  Most yeast cell lysates can be used as a positive control without induction or stimulation.  For ELISA a 1:1,000 to 1:5,000 dilution is recommended.   Researchers should determine optimal titers for other applications.
Purity/Specifity
This product is an IgG fraction antibody purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above.  Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
General Reference
Specific Reference
Integrating high-throughput genetic interaction mapping and high-content screening to explore yeast spindle morphogenesis. J Cell Biol. 2010 Jan 11;188(1):69-81. doi: 10.1083/jcb.200909013.
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Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC
Reacts With
yeast
Catalog Number
200-301-428

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
yeast
Catalog Number
200-401-428

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
human
Catalog Number
200-401-441

Product Type
Primary Antibodies;
Applications
ELISA, WB, IF
Reacts With
human
Catalog Number
200-401-491

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry