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Primary Antibodies  >  Ubiquitin and UBL Antibodies
 

Ubiquitin Antibody

Rabbit Polyclonal
Code

200-401-431
Size

500 µg
Price

$309.00
Availability

Ships next business day.
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Western blot of Ubiquitin.  Anti-Ubiquitin antibody, generated by immunization with Ubiquitin coupled to Rabbit IgG, was tested by western blot against total cell extract from yeast.  Dilution of the antibody between 1:200 and 1:1,000 showed strong reactivity with Ubiquitinated proteins.  In this blot the antibody was used at a 1:500 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 chemi-luminescence system was used for signal detection (Roche).  Other detection systems will yield similar results.
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Synonyms: FLJ25987 antibody, MGC8385 antibody, Polyubiquitin B antibody, RPS27A antibody, UBA52 antibody, UBA80 antibody, UBB antibody, UBC antibody, UBCEP1 antibody

Ubiquitin Antibody Properties

Description
Anti-UBIQUITIN (RABBIT) Antibody - 200-401-431
Host
Rabbit
Known Cross Reactivity
A Broad Range Of Species
Clonality
Polyclonal
Application
ELISA : 1:1,000 - 1:5,000
Western Blot : 1:200 - 1:1000
Immunochemistry: User Optimized
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
UBB
Download CofA
Lot#  

Ubiquitin Antibody Description

Background
Ubiquitin (Ub) is a small, 76-residue, protein (8.5 kDa) found both as free monomer and covalently attached to itself and other proteins in eukaryotic cells. Free Ub is a very compact and stable molecule that is easily refolded after being denatured. It is therefore recommended that for detection of free Ub on Westerns, the Tris-Tricine SDS-PAGE is used and nitrocellulose filters are autoclaved after the transfer and before blocking and addition of anti-Ub antibodies. The C-terminus of ubiquitin forms an isopeptide bond with the e-amino group of a lysine side chain in a target protein. In this way proteins can be covalently modified by the addition of ubiquitin which may alter the target protein's function. Monoubiquitination generally targets proteins for internalization, endocytosis and lysosomal degradation, or modifies the surface charge of histones and affects chromatin compaction.  Conjugation of ubiquitin (Ub) involves a three-step mechanism whereby specific enzymes (or enzyme complexes) activate and covalently link Ub to their substrates.
Immunogen
This purified antibody was prepared from rabbit serum after repeated immunizations with ubiquitin coupled to rabbit IgG using glutaraldehyde.
Immunogen Type
Native 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 with ubiquitin by ELISA and western blot. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation. For detection of free Ub by western blotting use Tris-Tricine SDS-PAGE and autoclaved nitrocellulose filters after the transfer and before blocking and addition of anti-Ub antibodies. Details on western blotting procedures are found in Mimnaugh et al., (1999 and 2002).
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 326, Gilbertsville, Pennsylvania, USA.
General Reference
Mimnaugh, E.G., Bovini, P. and Neckers, L. (1999) The measurement of ubiquitin and ubiquinated proteins. Electrophoresis 20:418-428. Wilkinson, K.D. (2000) Ubiquination and deubiquination: targeting of proteins for degradation by the proteasome. Cell Dev. Bio.11:141-148. Wilkinson, K. D., and Hochstrasser, M. (1998) in Ubiquitin and the Biology of the Cell (Peters, J. M. , Harris, J. R. , and Finley, D., eds) , pp. 99-125, Plenum Press, New York.
Specific Reference
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Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IP, ChIP
Reacts With
human
Catalog Number
200-301-174

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC
Reacts With
human
Catalog Number
200-401-888

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
human, mouse
Catalog Number
600-401-987

Product Type
Primary Antibodies;
Applications
ELISA, WB, IP
Reacts With
human
Catalog Number
600-401-991

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry