Revision 1

#9046Store at -20C

Cell Signaling Technology

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

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

28

SOURCE:

Rabbit

UniProt ID:

#P41227

Entrez-Gene Id:

8260

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50

Storage

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

Specificity / Sensitivity

ARD1A Antibody recognizes endogenous levels of total ARD1A protein. This antibody does not cross-react with the highly homologous ARD1B protein. This antibody also cross-reacts with a protein of unknown origin at 60 kDa.

Species Reactivity:

Human, Monkey

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val171 of human ARD1A protein. Antibodies are purified by protein A and peptide affinity chromatography.

Background

Protein acetylation is a common modification that occurs both at lysine residues within proteins (ε-amino acetylation) and multiple amino acid residues at the amino terminus of proteins (α-amino acetylation). The N-α-acetyltransferase ARD1 homolog A protein (ARD1A, also known as NAA10) and the highly homologous N-α-acetyltransferase ARD1 homolog B protein (ARD1B, also known as ARD2 or NAA11) are mutually exclusive catalytic subunits of the amino-terminal acetyltransferase complex (NatA) (1-3). This complex, which consists of either ARD1A or ARD1B and the N-α-acetyltransferase 15 (NAA15) auxiliary protein, localizes to ribosomes where it functions to acetylate Ser-, Ala-, Gly-, Thr-, Cys-, Pro-, and Val- amino termini after initiator methionine cleavage during protein translation (1-5). Like ε-amino acetylation, amino-terminal α-amino acetylation functions to regulate protein stability, activity, cellular localization, and protein-protein interactions (4,5). Defects in ARD1A have been shown to cause amino-terminal acetyltransferase deficiency (NATD), which results in severe delays and defects in postnatal growth (6).

In addition to functioning as amino-terminal acetyltransferases in the NatA complex, free ARD1A and ARD1B proteins regulate cell growth and differentiation through ε-amino acetylation of lysine residues in multiple target proteins, including the HIF-1α, β-catenin, and AP-1 transcription factors (7-9). ARD1A-mediated acetylation of HIF-1α at Lys532 under normoxic conditions enhances binding of VHL, leading to increased ubiquitination and degradation of HIF-1α and down-regulation of HIF-1α target genes involved in angiogenesis, apoptosis, cellular proliferation, and glucose metabolism (7). Decreased expression of ARD1A under hypoxic conditions contributes to the stabilization of HIF-1α and upregulation of target genes (7). ARD1A also promotes cell proliferation and tumorigenesis by acetylating and activating β-catenin and AP-1 transcription factors, leading to the stimulation of cyclin D1 expression (8,9). Interestingly, the acetyltransferase activity of ARD1A is regulated by autoacetylation at Lys136, which is required for the ability of ARD1A to promote proliferation and tumorigenesis (9). Research studies have shown that ARD1 proteins are over-expressed in multiple cancers, including breast, prostate, lung, and colorectal cancers (10-13).

  1. Arnesen, T. et al. (2005) Biochem J 386, 433-43.
  2. Arnesen, T. et al. (2006) BMC Biochem 7, 13.
  3. Pang, A.L. et al. (2009) Biol Reprod 81, 302-9.
  4. Van Damme, P. et al. (2011) FEBS J 278, 3822-34.
  5. Polevoda, B. and Sherman, F. (2000) J Biol Chem 275, 36479-82.
  6. Rope, A.F. et al. (2011) Am J Hum Genet 89, 28-43.
  7. Jeong, J.W. et al. (2002) Cell 111, 709-20.
  8. Lim, J.H. et al. (2006) Cancer Res 66, 10677-82.
  9. Seo, J.H. et al. (2010) Cancer Res 70, 4422-32.
  10. Arnesen, T. et al. (2005) Thyroid 15, 1131-6.
  11. Ren, T. et al. (2008) Cancer Lett 264, 83-92.
  12. Yu, M. et al. (2009) Oncol Rep 21, 909-15.
  13. Yu, M. et al. (2009) Cancer Invest 27, 978-83.

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 IP: Immunoprecipitation

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.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

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Revision 1
#9046

ARD1A Antibody

Western Blotting Image 1: ARD1A Antibody Expand Image
Western blot analysis of extracts from various cell lines using ARD1A Antibody.
Immunoprecipitation Image 1: ARD1A Antibody Expand Image
Immunoprecipitation of ARD1A from HeLa cell extracts, using Normal Rabbit IgG #2729 (lane 2) or ARD1A Antibody (lane 3). Lane 1 is 10% input. Western blot analysis was performed using ARD1A Antibody.