Immunofluorescence analysis of Phospho-FAK2 / PYK2 pTyr580 was done on 70% confluent log phase A549 cells treated with 50ng of PDGF for 10 minutes. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-FAK2 / PYK2 pTyr580 Rabbit Polyclonal Antibody (44636G) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing membranous localization. Panel e is untreated cell with no signal. Panel f is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human Pyk2 that contains tyrosines 579 and 580. The sequence is conserved in mouse and rat.|
|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 *|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|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.
Microglia Activate Migration of Glioma Cells through a Pyk2 Intracellular Pathway.
44-636G was used in western blot to examine how microglia promote glioma migration.
|Rolón-Reyes K,Kucheryavykh YV,Cubano LA,Inyushin M,Skatchkov SN,Eaton MJ,Harrison JK,Kucheryavykh LY||PloS one (10:null)||2015|
Collagen XV inhibits epithelial to mesenchymal transition in pancreatic adenocarcinoma cells.
44-636G was used in western blot to study COLXV in pancreatic adenocarcinoma cells.
|Clementz AG,Mutolo MJ,Leir SH,Morris KJ,Kucybala K,Harris H,Harris A||PloS one (8:null)||2013|
|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|
||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|