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Primary Antibodies  >  Ubiquitin and UBL Antibodies
 

SAE1 phospho S185 Antibody

Rabbit Polyclonal
Code

600-401-B24
Size

100 µg
Price

$309.00
Availability

Ships next business day.
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Western blot using Rockland's Rabbit anti-SAE1 pS185 antibody shows detection of phosphorylated SAE1.  Left lane  (-) contains 20 µg human HeLa whole cell protein.  Right lane (+) contains 20 µg human HeLa whole cell protein from cells pre-treated with phosphatase inhibitor cocktail to prevent dephos-phorylation of the target.  Proteins were separated on a 10% SDS-PAGE and transferred onto nitrocellulose.   After blocking with 5% milk-TBST 1 hr at room temperature, the membrane was probed with the primary antibody diluted to 2 µg/mL at room temperature for 3 hr followed by washes and reaction with HRP-conjugated secondary and ECL imaging.   Personal communication, Xin-Hua Feng, Baylor College of Medicine, Houstin, TX.
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Synonyms: SUMO E1 activating enzyme Ubiquitin-like 1 activating enzyme E1A UBLE1A AOS1 SAE1 SUA1

SAE1 phospho S185 Antibody Properties

Description
Anti-SUMO Activating Enzyme E1 (SAE1) pS185 (RABBIT) Antibody - 600-401-B24
Host
Rabbit
Target Species
Human
Known Cross Reactivity
human, bovine, mouse, rat, dog
Clonality
Polyclonal
Application
ELISA : 1:5,000 - 1:25,000
Western Blot : 2 µg/mL
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping condition
Dry Ice
Format
IgG
Label
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
SAE1
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Lot#  

SAE1 phospho S185 Antibody Description

Background
SUMO E1 activating enzyme (also called Ubiquitin-like 1 activating enzyme E1A, UBLE1A, AOS1, SAE1, and SUA1) with SAE2 (also known as UBA2) forms a heterodimeric (SAE1/SAE2) enzyme that activates the ubiquitin-like SUMO proteins (SUMO stands for Small Ubiquitin-like MOdifier.) The SAE1 (SUMO Activating Enzyme 1) subunit resembles the N-terminal half of yeast UBA1; the SAE2 (also called Uba2) subunit corresponds to the C-terminal part of yeast UBA1 and contains the active site cysteine. In the SUMO activation step, SAE1/SAE2 uses ATP to adenylate the C-terminal glycine of SUMO-1 (the first of the three different mammalian SUMO proteins) then forms a high-energy thioester bond between the C-terminal glycine and the active site cysteine in SAE2 (Uba2). In the conjugation step, the SUMO moiety is transferred from SAE1/SAE2 to the active site cysteine (Cys 93) of the SUMO conjugating enzyme (SUMO E2, Ubc9) forming a SUMO-E2 thioester complex.
Immunogen
This purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to a region surrounding S185 of the human SUMO Activating Enzyme E1 protein.
Immunogen Type
Peptide
Post Translational Modification
Phosphorylation
Storage Condition
Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. 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 antibody has been tested for use in ELISA and western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band at ~37 kDa in size corresponding to phosphorylated SAE1 protein by western blotting in the appropriate cell lysate or extract. This phospho-specific antibody reacts with human SAE1 pS185 and shows minimal reactivity by ELISA against the non-phosphorylated form of the immunizing peptide.
Purity/Specifity
This purified antibody is directed against human SUMO Activating Enzyme E1 protein. The product was purified from monospecific antiserum by affinity chromatography. This antibody is specific for human SAE1 protein phosphorylated at S185. A BLAST analysis using the sequence of the immunizing peptide was used to suggest that this antibody would react with SUMO Activating Enzyme E1 protein from human (100%), bovine, dog, chimpanzee (96%), mouse (93%), and rat (92%) based on a high degree of sequence homology. Cross reactivity against this protein from other sources has not been determined.
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 326, Gilbertsville, Pennsylvania, USA.
General Reference
Wrighton KH,and Feng XH. (2006). Uba2. AfCS-Nature Molecule Pages. doi:10.1038/mp.a003681.01 Lois,L.M. and Lima,C.D. (2005) Structures of the SUMO E1 provide mechanistic insights into SUMO activation and E2 recruitment to E1. EMBO J. 24 (3), 439-451 Desterro,J.M., Rodriguez,M.S., Kemp,G.D. and Hay,R.T. (1999) Identification of the enzyme required for activation of the small ubiquitin-like protein SUMO-1. J. Biol. Chem. 274 (15), 10618-10624. Gong,L., Li,B., Millas,S. and Yeh,E.T. (1999) Molecular cloning and characterization of human AOS1 and UBA2, components of the sentrin-activating enzyme complex. FEBS Lett. 448 (1), 185-189.
Specific Reference
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Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IF, FC
Reacts With
human, mouse, rat, monkey
Catalog Number
200-301-268

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IF
Reacts With
human
Catalog Number
600-401-879

Product Type
Secondary Antibodies;
Applications
ELISA, WB, IHC
Catalog Number
610-4302

Product Type
Secondary Antibodies;
Applications
ELISA, WB, IHC
Catalog Number
611-1302

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry