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Primary Antibodies  >  Ubiquitin and UBL Antibodies
 

Ubiquitin Activating Enzyme E1 Antibody

Rabbit Polyclonal
Code

200-401-888
Size

500 µg
Price

$309.00
Availability

Ships next business day.
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Rockland's Affinity Purified anti-Ubiquitin Activating Enzyme antibody was used at a 10 µg/ml to detect UBE1 in a variety of tissues including adrenal, breast, colon (epithelium), kidney, liver, lung (respiratory epithelium), ovary (oocyte and endothelium), pancreas (islet and exocrine), placenta, prostate (epithelium), skin (epithelium), spleen (lymphocytes), stomach (chief), testis, thymus, tonsil, and uterus (glandular, stroma).  In many cells a punctate nuclear staining was observed.  Other cells showed both cytoplasmic and nuclear staining.  This image shows UBE1 staining of human lung tissue.  Tissue was formalin-fixed and paraffin embedded.  Personal Communi-cation, Tina Roush, LifeSpanBiosciences, Seattle, WA.
Western blot using Rockland's purified anti-Ubiquitin Activating Enzyme (E1) antibody shows detection of a band at ~118 kDa corres-ponding to UBE1 (lane 1 800 nm channel).   Approximately 35 µg of an A431 whole cell lysate was separated on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred onto nitrocellulose.   After blocking the membrane was probed with the primary antibody diluted to 1:1,000.  Incubation was for 2 h at room temperature followed by washes and reaction with a 1:10,000 dilution of IRDye™800 conjugated Gt-a-Rabbit IgG [H&L] MX10 (611-132-122) for 45 min at room temperature.  Molecular weight markers are shown in lane 2 (700 nm channel).  IRDye™800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc.  Other detection systems will yield similar results.
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Synonyms: A1S9 antibody, A1S9 protein antibody, A1S9T and BN75 temperature sensitivity complementing antibody, A1S9T antibody, A1ST antibody, GXP 1 antibody

Ubiquitin Activating Enzyme E1 Antibody Properties

Description
Anti-Ubiquitin Activating Enzyme E1 [RABBIT) Antibody - 200-401-888
Host
Rabbit
Target Species
Human
Known Cross Reactivity
human
Clonality
Polyclonal
Application
ELISA : 1:2,000 - 1:10,000
Western Blot : 1:1,000 - 1:5,000
Immunochemistry: 2 mg/ml - 20 µg/ml
Other Dilution: User Optimized
Physical State
Lyophilized
Shipping condition
Ambient
Format
IgG
Label
Concentration
5.0 mg/mL by UV absorbance at 280 nm
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Reconstitution Volume
100 µL
Reconstitution Buffer
Restore with deionized water (or equivalent)
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
UBA1
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Lot#  

Ubiquitin Activating Enzyme E1 Antibody Description

Background
Ubiquitin Activating Enzyme (E1), also known as A1S9 and UBE1, is responsible for the first step in ubiquitin-protein isopeptide bond formation.  E1 catalyzes the activation of the C-terminal carboxyl group of ubiquitin by forming a high-energy thioester bond in an ATP-dependent manner. UBE1 is monomeric and contains two active sites within the E1 molecule, allowing it to bind two ubiquitin moieties at a time, with a new ubiquitin forming an adenylate intermediate as the previous one is transferred to the thiol site.  Alternative splicing results in 2 transcript variants encoding the same protein, but with different 5' UTR. Isoform 1 has a different 5' noncoding exon compared to isoform 2.  Both variants encode the same protein.
Immunogen
Anti-Ubiquitin Activating Enzyme E1 antibody was prepared from whole rabbit serum produced by repeated immunizations with a recombinant protein corresponding to full length Human Ubiquitin Activating Enzyme E1.
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 antibody has been tested for use in ELISA, immunohistochemistry and western blot.  Specific conditions for reactivity should be optimized by the end user.  Expect a band at ~118 kDa in size corresponding to UBE1 by western blotting in the appropriate cell lysate or extract.
Purity/Specifity
This purified antibody is directed against human Ubiquitin Activating Enzyme E1 protein.  The product was purified from monospecific antiserum by a multi step procedure.  A BLAST analysis was used to suggest that this antibody would react with Ubiquitin Activating Enzyme E1 protein from human (100%) rabbit (96%), mouse (95%), rat (95%) and dog (93%) 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
Beausoleil,S.A., Jedrychowski,M., Schwartz,D., Elias,J.E., Villen,J., Li,J., Cohn,M.A., Cantley,L.C. and Gygi,S.P. (2004) Large-scale characterization of HeLa cell nuclear phosphoproteins. Proc. Natl. Acad. Sci. U.S.A. 101 (33), 12130-12135. Saito,Y., Doi,K., Yamagishi,N., Ishihara,K. and Hatayama,T. (2004) Screening of Hsp105alpha-binding proteins using yeast and bacterial two-hybrid systems. Biochem. Biophys. Res. Commun. 314 (2), 396-402. Bres,V., et al. (2003) A non-proteolytic role for ubiquitin in Tat-mediated transactivation of the HIV-1 promoter. Nat. Cell Biol. 5 (8), 754-761.
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
Secondary Antibodies;
Applications
ELISA, WB, IHC
Catalog Number
610-4302

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

Product Type
Supporting Reagents;
Applications
ELISA, FLISA, WB, IHC, IF, IP, FC
Catalog Number
B304

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry