Antibody-Peptide Competition. Extracts of Jurkat cells treated with 5 nM SDF-1alpha for 20 minutes were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature, then incubated with the LIMK1/2 [pYpT507/508] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphotyrosine-containing peptide (3), a generic phosphothreonine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F(ab and quote;)2 anti-rabbit IgG HRP conjugate (Cat. no. ALI4404) and signals were detected using the Pierce SuperSignal™ method. The data show that only the phosphopeptide corresponding to LIMK1/2 [pYpT507/508] blocks the antibody signal, demonstrating the specificity of the antibody.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human LIMK1 that contains tyrosine 507 and threonine 508 (tyrosine 504 and threonine 505 in LIMK2). The LIMK1 sequence is conserved in mouse and rat, and the LIMK2 sequence is conserved in mouse, rat, and chicken.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
LIMK1, a member of the Ser/Thr protein kinase family, may be a component of an intracellular signaling pathway and may be involved in brain development. It phosphorylates and inactivates the actin binding/depolymerizing factor cofilin and induces actin cytoskeletal changes. The LIM domain interacts with the cytoplasmic domain of NRG1, and this cytoplasmic protein also binds ROCK1, whic phosphorylates LIMK1 on serine and/or threonine residues. Highest expression occurs in both adult and fetal nervous systems. It is detected ubiquitously throughout the different regions of adult brain, with highest levels in the cerebral cortex, and is expressed to a lesser extent in heart and skeletal muscle. Haploinsufficiency of LIMK1 may be the cause of certain cardiovascular and musculo-skeletal abnormalities observed in Williams-Beuren syndrome (WBS), a rare developmental disorder. It is a contiguous gene deletion syndrome involving genes from chromosome band 7q11.23. This protein contains 2 LIM zinc-binding domains and 1 PDZ/DHR domain.
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