Revision 1

#14321Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

60

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q13564

Entrez-Gene Id:

8883

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

NAE1/APPBP1 (D9I4Z) Rabbit mAb recognizes endogenous levels of total NAE1/APPBP1 protein. This antibody does not cross-react with E1 activating enzymes for either ubiquitin or other ubiquitin-like proteins.

Species Reactivity:

Human, Mouse, Rat, Monkey

Species predicted to react based on 100% sequence homology

Bovine, Dog

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala179 of human NAE1/APPBP1 protein.

Background

Similar to ubiquitin, NEDD8 is covalently linked to target proteins through an enzymatic cascade composed of NEDD8-specific E1 (activating)- and E2 (conjugating)-enzymes (1,2). The E2 ligase specific for NEDD8 is Ubc12 (3-5). Ubc12 forms a heterodimeric conjugate with NEDD8 in order to catalyze the transfer of NEDD8 from E1 to lysine side chains of target proteins (1,2). Well known targets of NEDD8 are cullin-based RING E3 ligases. Neddylation of cullin isoforms activates the related ubiquitin E3 complex by promoting its interaction with a cognate ubiquitin-E2 ligase (6-7). Neddylation of Cul-1 complexes containing βTrCP and SKP2 has been shown to be required for controlling the stability of important signaling targets such as IκB, NF-κB, and p27 Kip (8-10), thereby regulating cell cycle progression, signaling cascades, and developmental programming processes (11).
NAE1/APPBP1 (NEDD8-Activating Enzyme 1/Amyloid Beta Precursor Protein Binding Protein 1) exists in a heterodimeric complex with UBA3. This complex functions as an E1 NEDD8-activating enzyme, which utilizes ATP to adenylate the C-terminal glycine of NEDD8 (12-14). Research studies suggest that inhibition of the APPBP1-UBA3 complex may be of therapeutic value for the treatment of human cancers (15,16).

  1. Huang, D.T. et al. (2007) Nature 445, 394-8.
  2. Huang, D.T. et al. (2005) Mol Cell 17, 341-50.
  3. Liakopoulos, D. et al. (1998) EMBO J 17, 2208-14.
  4. Gong, L. and Yeh, E.T. (1999) J Biol Chem 274, 12036-42.
  5. Wada, H. et al. (2000) J Biol Chem 275, 17008-15.
  6. Sakata, E. et al. (2007) Nat Struct Mol Biol 14, 167-8.
  7. Kawakami, T. et al. (2001) EMBO J 20, 4003-12.
  8. Podust, V.N. et al. (2000) Proc Natl Acad Sci USA 97, 4579-84.
  9. Wu, K. et al. (2002) J Biol Chem 277, 516-27.
  10. Amir, R.E. et al. (2002) J Biol Chem 277, 23253-9.
  11. Herrmann, J. et al. (2007) Circ Res 100, 1276-91.
  12. Walden, H. et al. (2003) Mol Cell 12, 1427-37.
  13. Walden, H. et al. (2003) Nature 422, 330-4.
  14. Huang, D.T. et al. (2004) Nat Struct Mol Biol 11, 927-35.
  15. Soucy, T.A. et al. (2009) Nature 458, 732-6.
  16. Swords, R.T. et al. (2010) Blood 115, 3796-800.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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Revision 1
#14321

NAE1/APPBP1 (D9I4Z) Rabbit mAb

Western Blotting Image 1: NAE1/APPBP1 (D9I4Z) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using NAE1/APPBP1 (D9I4Z) Rabbit mAb.
Western Blotting Image 2: NAE1/APPBP1 (D9I4Z) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human NAE1/APPB1 protein (hNAE1-Myc/DDK; +), using NAE1/APPBP1 (D9I4Z) Rabbit mAb.