Immunofluorescence analysis of PYK2 [pY402] was done on 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with PYK2 [pY402] Rabbit Polyclonal Antibody (44618G) at 1ug/mL in 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 cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Chicken, Human, Mouse, Rat|
|Published species reactivity||Tag, Rat, Human, Mouse, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human Pyk2 that contains tyrosine 402. The sequence is conserved in 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))||1:10-1:50|
|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.
PYK2 selectively mediates signals for growth versus differentiation in response to stretch of spontaneously active vascular smooth muscle.
44-618G was used in western blot to examine the role of the calcium- and integrin-activated proline-rich tyrosine kinase 2 in stretch-induced responses of the rat portal vein.
|Bhattachariya A,Turczy¿ska KM,Grossi M,Nordström I,Buckbinder L,Albinsson S,Hellstrand P||Physiological reports (2:null)||2014|
|Biophysical stimulation induces demyelination via an integrin-dependent mechanism.||Lin MY,Frieboes LS,Forootan M,Palispis WA,Mozaffar T,Jafari M,Steward O,Gall CM,Gupta R||Annals of neurology (72:112)||2012|
|Human||Not Cited||ICAM-1-mediated, Src- and Pyk2-dependent vascular endothelial cadherin tyrosine phosphorylation is required for leukocyte transendothelial migration.||Allingham MJ,van Buul JD,Burridge K||Journal of immunology (Baltimore, Md. : 1950) (179:4053)||2007|
|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|
|Not Applicable||Not Cited||
Activation of FAK is necessary for the osteogenic differentiation of human mesenchymal stem cells on laminin-5.
44-618G was used in western blot to report that FAK is an important mediator of laminin-5-induced osteogenic differentiation of human mesenchymal stem cells
|Salasznyk RM,Klees RF,Boskey A,Plopper GE||Journal of cellular biochemistry (100:499)||2007|
|Not Applicable||Not Cited||
Focal adhesion kinase signaling pathways regulate the osteogenic differentiation of human mesenchymal stem cells.
44-618G was used in western blot to investigate FAK signaling in the regulation of extracellular matrix-induced osteogenic differentiation of human mesenchymal stem cells
|Salasznyk RM,Klees RF,Williams WA,Boskey A,Plopper GE||Experimental cell research (313:22)||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|
|Mouse||Not Cited||Role of PYK2 in the development of obesity and insulin resistance.||Yu Y,Ross SA,Halseth AE,Hollenbach PW,Hill RJ,Gulve EA,Bond BR||Biochemical and biophysical research communications (334:1085)||2005|
|Human||Not Cited||Focal adhesion kinase plays a pivotal role in herpes simplex virus entry.||Cheshenko N,Liu W,Satlin LM,Herold BC||The Journal of biological chemistry (280:31116)||2005|
|Rat||Not Cited||PYK2 regulates SERCA2 gene expression in neonatal rat ventricular myocytes.||Heidkamp MC,Scully BT,Vijayan K,Engman SJ,Szotek EL,Samarel AM||American journal of physiology. Cell physiology (289:C471)||2005|
|Human||Not Cited||Proline-rich tyrosine kinase 2 (Pyk2) mediates vascular endothelial-cadherin-based cell-cell adhesion by regulating beta-catenin tyrosine phosphorylation.||van Buul JD,Anthony EC,Fernandez-Borja M,Burridge K,Hordijk PL||The Journal of biological chemistry (280:21129)||2005|
|Rat||Not Cited||Glucose activates protein kinase C-zeta /lambda through proline-rich tyrosine kinase-2, extracellular signal-regulated kinase, and phospholipase D: a novel mechanism for activating glucose transporter translocation.||Bandyopadhyay G,Sajan MP,Kanoh Y,Standaert ML,Quon MJ,Reed BC,Dikic I,Farese RV||The Journal of biological chemistry (276:35537)||2001|
|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|
CD28 and CD3 have complementary roles in T-cell traction forces.
44-618G was used in immunocytochemistry to investigate the effect of mechanical force on the T cell receptor.
|Bashour KT,Gondarenko A,Chen H,Shen K,Liu X,Huse M,Hone JC,Kam LC||Proceedings of the National Academy of Sciences of the United States of America (111:2241)||2014|
||Biophysical stimulation induces demyelination via an integrin-dependent mechanism.||Lin MY,Frieboes LS,Forootan M,Palispis WA,Mozaffar T,Jafari M,Steward O,Gall CM,Gupta R||Annals of neurology (72:112)||2012|
Knock-in mutation reveals an essential role for focal adhesion kinase activity in blood vessel morphogenesis and cell motility-polarity but not cell proliferation.
44-618G was used in western blot to determine the role of FAK activity during development
|Lim ST,Chen XL,Tomar A,Miller NL,Yoo J,Schlaepfer DD||The Journal of biological chemistry (285:21526)||2010|
Integrin dynamics produce a delayed stage of long-term potentiation and memory consolidation.
44-618G was used in immunohistochemistry to study the role of beta integrin in long-term potentiation and the consolidation of memory.
|Babayan AH,Kramár EA,Barrett RM,Jafari M,Häettig J,Chen LY,Rex CS,Lauterborn JC,Wood MA,Gall CM,Lynch G||The Journal of neuroscience : the official journal of the Society for Neuroscience (32:12854)||2012|
||ICAM-1-mediated, Src- and Pyk2-dependent vascular endothelial cadherin tyrosine phosphorylation is required for leukocyte transendothelial migration.||Allingham MJ,van Buul JD,Burridge K||Journal of immunology (Baltimore, Md. : 1950) (179:4053)||2007|
||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|
||Proline-rich tyrosine kinase 2 (Pyk2) mediates vascular endothelial-cadherin-based cell-cell adhesion by regulating beta-catenin tyrosine phosphorylation.||van Buul JD,Anthony EC,Fernandez-Borja M,Burridge K,Hordijk PL||The Journal of biological chemistry (280:21129)||2005|