Peptide Competition. Extracts of CEF cells mock transfected (1) or transfected with Pyk2 and plated on fibronectin (2-7) 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 Pyk2 [pY580] antibody for two hours at room temperature in a 1% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), the phosphopeptide corresponding to Pyk2 [pY579] (4), a phosphopeptide derived from the corresponding region of FAK (5), a generic phosphotyrosine-containing peptide (6) or the phosphopeptide immunogen (7). 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 Pyk2 [pY580] completely blocks the antibody signal, demonstrating the specificity of the antibody.
|Tested species reactivity||Human|
|Published species reactivity||Rat, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human Pyk2 that contains tyrosine 580. The sequence is conserved in mouse and rat.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (IHC)||Assay Dependent|
|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.
This gene encodes a cytoplasmic protein tyrosine kinase which is involved in calcium-induced regulation of ion channels and activation of the map kinase signaling pathway. The encoded protein may represent an important signaling intermediate between neuropeptide-activated receptors or neurotransmitters that increase calcium flux and the downstream signals that regulate neuronal activity. The encoded protein undergoes rapid tyrosine phosphorylation and activation in response to increases in the intracellular calcium concentration, nicotinic acetylcholine receptor activation, membrane depolarization, or protein kinase C activation. This protein has been shown to bind CRK-associated substrate, nephrocystin, GTPase regulator associated with FAK, and the SH2 domain of GRB2. The encoded protein is a member of the FAK subfamily of protein tyrosine kinases but lacks significant sequence similarity to kinases from other subfamilies. Four transcript variants encoding two different isoforms have been found for this gene.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Human||Not Cited||The role of proline-rich protein tyrosine kinase 2 in differentiation-dependent signaling in human epidermal keratinocytes.||Schindler EM,Baumgartner M,Gribben EM,Li L,Efimova T||The Journal of investigative dermatology (127:1094)||2007|
|Mouse||Not Cited||Analyzing FAK and Pyk2 in early integrin signaling events.||Bernard-Trifilo JA,Lim ST,Hou S,Schlaepfer DD,Ilic D||Current protocols in cell biology (Chapter 14:null)||2006|
|Rat||Not Cited||Implication of phospholipase D2 in oxidant-induced phosphoinositide 3-kinase signaling via Pyk2 activation in PC12 cells.||Banno Y,Ohguchi K,Matsumoto N,Koda M,Ueda M,Hara A,Dikic I,Nozawa Y||The Journal of biological chemistry (280:16319)||2005|
|Rat||Not Cited||Salicylate Inhibits Phosphorylation of the Nonreceptor Tyrosine Kinases, Proline-Rich Tyrosine Kinase 2 and c-Src.||Wang Z,Brecher P||Hypertension (Dallas, Tex. : 1979) (37:148)||2001|
||Analyzing FAK and Pyk2 in early integrin signaling events.||Bernard-Trifilo JA,Lim ST,Hou S,Schlaepfer DD,Ilic D||Current protocols in cell biology (Chapter 14:null)||2006|
||Implication of phospholipase D2 in oxidant-induced phosphoinositide 3-kinase signaling via Pyk2 activation in PC12 cells.||Banno Y,Ohguchi K,Matsumoto N,Koda M,Ueda M,Hara A,Dikic I,Nozawa Y||The Journal of biological chemistry (280:16319)||2005|