Human NEDD8 Antibody

(Catalog # AP025)

Specificity: Rabbit polyclonal antibody against human Ubiquitin-like protein NEDD8.

Antigen used for immunizations: Synthesized MAP (multiple antigen protein) containing eight asymmetric branches of the amino acid residues 8 to 22 (LTGKEIEIDIEPTDK) of the human Ubiquitin-like protein NEDD8 [Accession #: Q15843].

Storage Condition: Whole serum with 0.1% NaN3.

Recommendations on use:

Immunofluorescence:

Not applicable.

Western blotting: Recommended:

1. Run and transfer SDS-PAGE for blotting assay.

2. Rinse the transferred membrane with ddWater and let it dry.

3. Block transfer membrane with 5% skim milk, 02% sodium azide in 10mM TBS, incubate at  37ºC for at least 2 hours or  4ºC overnight. Then use 0.05% Tween 20 in TBS to wash the membrane.

4. 60 minutes primary antibody incubates at 37ºC (Ab is serially diluted in 5% FBS/TBS).

5. Use 0.05% Tween 20 in TBS to wash the membrane three times.

6. Add 2nd Ab (Goat anti-rabbit IgG 3000x dilution in 5% FBS/TBS) and incubate at 37ºC for one hour.

7. Use 0.05% Tween 20 in TBS to wash the membrane three times. Then wash the membrane with ddWater. Soak the membrane in BCIP/NBT substrate solution, after the color developed, wash with ddWater and let it dry.

(Data Will Be Shown When Available.)

Storage and stability: Store unopened antibody at 4ºC or lower. Under these conditions, there is no significant loss in product performance up to one year from the purchase date. The reconstituted antibody is stable for at least two months when stored at 4ºC. For long term storage, it is recommended that aliquots of the antibody are frozen at -20ºC (frost-free freezers are not recommended). Repeated freezing and thawing must be avoided. Prepare working dilutions on the day of use.

Legal consideration: FOR RESEARCH USE ONLY.

Reference:

Osaka,F., Kawasaki,H., Aida,N., Saeki,M., Chiba,T., Kawashima,S., Tanaka,K. and Kato,S. A new NEDD8-ligating system for cullin-4A. Genes Dev. 12 (15), 2263-2268 (1998) MEDLINE   98361870  PUBMED   9694792
　

Whitby,F.G., Xia,G., Pickart,C.M. and Hill,C.P. Crystal structure of the human ubiquitin-like protein NEDD8 and interactions with ubiquitin pathway enzymes. J. Biol. Chem. 273 (52), 34983-34991 (1998) MEDLINE   99074275   PUBMED   9857030
　

Kito,K., Yeh,E.T. and Kamitani,T. NUB1, a NEDD8-interacting protein, is induced by interferon and down-regulates the NEDD8 expression. J. Biol. Chem. 276 (23), 20603-20609 (2001)  MEDLINE   21282873   PUBMED   11259415
　

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