Technical Data
A0921-97
Afadin, phosphorylated (Ser1718) (ALL1-fused Gene from Chromosome 6 Protein, AF6, FLJ34371, MLLT4, Protein AF-6, RP3-431P23.3)
Description:
In multicellular organisms, intercellular junctions play essential roles in tissue integrity and maintenance of cell polarity. Tight junctions (TJs) form a continuous barrier to fluids across the epithelium and endothelium (reviewed in 1). Adherens junctions (AJs) are dynamic structures that form cell-cell contacts linking cells into a continuous sheet (reviewed in 2). The actin filament-binding protein, Afadin, binds to nectin forming a connection to the actin cytoskeleton (3). AJs are formed when nectin assembles cadherin at the cell-cell adhesion site and these
junctions are then involved in the formation and maintenance of TJs (4, 5). Afadin has two splice variants: l-afadin, which is ubiquitously expressed, and s-afadin, which is expressed
predominantly in neural tissue. s-Afadin is a shorter form lacking one of the three proline-rich regions found in l-afadin as well the carboxyl-terminal F-actin binding region (6). Human s-afadin is identical to AF-6, the ALL-1 fusion partner involved in acute myeloid leukemias (7). Recent
work has also shown that Afadin is involved in controlling the directionality of cell movement when it is localized at the leading edge of moving cells (8, 9). Phosphorylation at Ser1718 was discovered using an Akt substrate antibody.

Applications:
Suitable for use in Western Blot. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000 Incubate membrane with diluted antibody in 5% BSA, 1X TBS, 0.1% Tween-20 at 4C with gentle shaking, overnight.
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
100ul-20CBlue IceHumanRabbit
Concentration:
Not determined
Immunogen:
Synthetic phosphopeptide corresponding to residues surrounding Ser1718 of human afadin.
Purity:
Purified by Protein A affinity chromatography.
Form
Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA, 50% glycerol.
Specificity:
Recognizes endogenous levels of l-afadin protein only when phosphorylated at serine 1718. Species Crossreactivity: canine, monkey, mouse, rat.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
(1) Shin, K. et al. (2006) Annu Rev Cell Dev Biol 22, 207-35. (2) Harris, T.J. and Tepass, U. (2010) Nat Rev Mol Cell Biol 11, 502-14. (3) Ikeda, W. et al. (1999) J Cell Biol 146, 1117-32. (4) Sato, T. et al. (2006) J Biol Chem 281, 5288-99. (5) Ooshio, T. et al. (2007) J Cell Sci 120, 2352-65. (6) Mandai, K. et al. (1997) J Cell Biol 139, 517-28. (7) Prasad, R. et al. (1993) Cancer Res 53, 5624-8. (8) Miyata, M. et al. (2009) J Cell Sci 122, 4319-29. (9) Miyata, M. et al. (2009) J Biol Chem 284, 24595-609.