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Western blot - Nedd8 antibody (ab4751)
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Western blot - Nedd8 antibody (ab4751)
(enlarge)
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Product Name
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Nedd8 antibody
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See all NEDD8 antibodies (9)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to Nedd8
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Immunogen
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Recombinant full length protein (S. cerevisiae).
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Reacts with
(species key)
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Sc
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Tested applications
(see key)
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ELISA, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ELISA: 1/2000 - 1/10000.
WB: 1/1000. Detects a band of approximately 9.7 kDa.
Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
This antibody using the specified conditions may recognize other prominent intrinsic bands (UBLs or conjugates). Other intrinsic bands are readily detectable at lower dilutions.
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Positive control
(see definition)
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Most yeast cell lysates can be used as a positive control without induction or stimulation.
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Cellular localization
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Nuclear
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Research areas
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Cell Biology >> Proteolysis / Ubiquitin >> Proteasome / Ubiquitin >> Ub-like Proteins
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Relevance
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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), Apg12 and Hub1. 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 proteolytically processed or conjugated to other proteins. Rub1 and the corresponding human homolog NEDD8 are activated by the E1 ubiquitin activating enzyme UBA2, that forms isopeptide linkages between thio esters. NEDD8 shows 80% homology to ubiquitin. The best known targets of Rub1 modification are members of the cullin family. Cullins are subunits of an E3 ubiquitin ligase complex called the Skp1 /Cul1/Cdc53 F box (SCF). The SCF promotes transfer of ubiquitin from a ubiquitin conjugating enzyme (E2) to the target protein. Rub1 modification may regulate SCF function or localization.
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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IgG fraction
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Storage buffer
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Preservative: 0.01% Sodium Azide
Constituents: 0.2M Potassium Phosphate, 0.15M Sodium Chloride. pH 7.2 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Purification notes
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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.
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Form
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Liquid
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Concentration
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1.000 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Nedd8 antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for Nedd8 antibody images, references, products related to ab4751 and other tools.
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Nedd8 antibody images:
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 Western blot - Nedd8 antibody (ab4751)
ab4751 tested by immunoblot against a yeast cell lysate. A dilution of the antibody between 1:200 and 1:1,000 will show strong reactivity specifically with free Rub1 protein (indicated by arrow) and Rub1 conjugates. 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 Donkey polyclonal to Rabbit IgG (HRP) (ab7083) for 1 hour at room temperature. A chemiluminescence system was used for signal detection (Roche).
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Western blot - Nedd8 antibody (ab4751)
Immunoblot of Rub1 fusion protein. Anti-Rub1 antibody generated by immunization with recombinant yeast Rub1 was tested by immunoblot against yeast lysates expressing the Rub1-GFP fusion protein and other UBL fusion proteins. All UBLs possess limited homology to Ubiquitin and to each other, therefore it is important to know the degree of reactivity of each antibody against each UBL. Panel A shows total protein staining using ponceau. Panel B shows positions of free GFP or GFP containing recombinant proteins present in each lysate preparation after reaction with a 1:1,000 dilution of Goat polyclonal to GFP (ab6673) followed by reaction with a 1:15,000 dilution of Donkey polyclonal to Goat IgG (ab7125). Panel C shows specific reaction with Rub1 using a 1:1,000 dilution of ab4751 followed by reaction with a 1:15,000 dilution of Goat polyclonal to Rabbit IgG (HRP) (ab7090). All primary antibodies were diluted in TTBS buffer supplemented with 5% non-fat milk and incubated with the membranes overnight at 4° C. Yeast lysate proteins were separated by SDS-PAGE using a 15% gel. This data indicates that anti-Rub1 is highly specific and does not cross react with other UBLs, however, in other experiments (not shown) some cross reactivity is observed against ubiquitin. Include an anti-ubiquitin control any experiment where the specific detection of Rub1 is uncertain. Note that two bands are detected by anti-GFP indicating that most Rub1-GFP is processed in yeast. A chemiluminescence system was used for signal detection (Roche). Data contributed by M. Malakhov, www.lifesensors.com, personal communication.
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Search PubMed (MEDLINE) for references to NEDD8
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