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Primary Antibodies
Dab 1 (pY220) pAb Antibody
Background
Disabled 1 (Dab1) is an 80 kDa protein that is encoded by the Disabled-1 gene locus which is mutated in scrambler and yotari mutant mice. Phenotypically, the mutation of this gene produces motor defects and ataxia, disruption of neuronal migration, and severe cerebellar hypoplasia. Dab1 is an intracellular adapter protein that functions in downstream signaling events initiated by the secreted protein reelin. Dab1 contains a phosphotyrosine binding (PTB) domain in the amino terminus. Tyrosine phosphorylation of Dab1 is increased by reelin binding to the Very Low Density Lipoprotein Receptor (VLDLR) and Apolipoprotein E Receptor 2 (ApoER2) through stimulation of Src family kinases (e.g., Fyn). Src family kinase and c-Abl activities are themselves then stimulated by binding to tyrosine phosphorylated Dab1. Dab1 also mediates activation of Akt (PKB) by reelin resulting in inhibition of glycogen synthase kinase 3β (GSK-3β) and decreased phosphorylation of the microtubule-associated protein, Tau. Dab1 tyrosine 220 is a major site for reelin-induced Src family kinase-mediated phosphorylation in embryonic neurons.
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Description
Lysates prepared from HEK293 cells transfected with wild-type (1, 2, 5, 6) or Y220F mutant mouse Dab1 (3, 4, 7, 8) and left untreated (-) (1, 3, 5, 7) or treated with H2O2 (+) (2, 4, 6, 8) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4˚C, and incubated with a Dab1 pan antibody (1-4) or Dab1 [pY220] antibody (5-8) for two hours at room temperature in a 3% BSA-TBST buffer. After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG HRP-conjugated and signals were detected using the Pierce SuperSignal™ method.
The data show H2O2 stimulation of the phospho-Dab1 signal only in wild type Dab1-expressing cells. The Dab1 pan antibody reactivity confirms that similar levels of Dab1 protein were present in all cell lysates. These results demonstrate the specificity of the Dab1 [pY220] antibody for this site on the Dab1 protein.
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Source
Dab1 antibody was produced against a chemically synthesized phosphopeptide derived from the region of human Dab1 that contains tyrosine 220. The sequence is conserved in mouse, rat and chicken.
Purification
Dab1 antibody was purified from rabbit serum by Protein A Sepharose affinity chromatography.
Clonality / Clone
This is a polyclonal antibody.
Host
Dab 1 (pY220) pAb antibody was raised in rabbit.
Please use anti-rabbit secondary antibodies.
Specificity
Mouse (100% homologous) Dab1. Human, rat and chicken (100%) Dab1 have not been tested, but are expected to react.
Tested Application
WB
Buffer
Rabbit polyclonal immunoglobulin in Dulbecco’s phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), with 1.0 mg/mL BSA (IgG, protease free) as a carrier.
Storage
Store at -80˚C. Upon initial thawing, we recommend that this vial be briefly centrifuged to ensure recovery of the entire volume. The antibody should then be apportioned into working aliquots and stored at -80˚C. Avoid repeated freeze/thaw cycles.
Positive Control
- Human Embryonic Kidney (HEK293) cells transfected with wild-type mouse Dab1 protein.
Species Reactivity
H, M, R, C
Formulation
Rabbit polyclonal immunoglobulin in Dulbecco’s phosphate buffered saline (without Mg2+ and Ca2+), pH 7.3 (+/- 0.1), with 1.0 mg/mL BSA (IgG, protease free) as a carrier.
Short Description
Dab 1 (pY220) pAb
This product belongs to the following categories:
References- Arnaud, L., et al. (2003) Fyn tyrosine kinase is a critical regulator of disabled-1 during brain development. Curr. Biol. 13(1):9-17.
- Beffert, U., et al. (2002) Reelin-mediated signaling locally regulates protein kinase B/Akt and glycogen synthase kinase 3?. J. Biol. Chem. 277(51):49958-49964.
- Howell, B.W., et al. (2000) Dab1 tyrosine phosphorylation sites relay positional signals during mouse brain development. Curr. Biol. 10(15):877-885.
- Homayouni, R., et al. (1999) Disabled-1 binds to the cytoplasmic domain of amyloid precursor-like protein 1. J Neurosci. 19(17):7507-7515.
Datasheet 0801-W
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