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Immunofluorescence analysis of Phospho-Paxillin pTyr31 was done on 70% confluent log phase NIH/3T3 cells were treated with 50ng of PDGF for 10minutes and 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-Paxillin pTyr31 Rabbit Polyclonal Antibody (44720G) 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 cytoplasmic localization. Panel e is untreated cells with less signal. Panel f is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Tag, Rat, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human paxillin that contains tyrosine 31.|
|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 *|
|Immunocytochemistry (ICC)||2-3 ug/ml|
|Immunofluorescence (IF)||2-3 ug/ml|
|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.
Paxillin is a focal adhesion protein and a substrate for several tyrosine kinases such as src, FAK, and p210BRC/ABL. The tyrosine phosphorylation of paxillin is affected by conditions that change cell-cell adhesion. Paxillin associates tightly with FAK and Crk through its SH2 domain. This interaction is independent of the extracellular matrix. Although paxillin was initially discovered in fibroblasts, its phosphorylation may also be important during neurite extension during differentiation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|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|
|Mouse||Not Cited||Governing epidermal homeostasis by coupling cell-cell adhesion to integrin and growth factor signaling, proliferation, and apoptosis.||Livshits G,Kobielak A,Fuchs E||Proceedings of the National Academy of Sciences of the United States of America (109:4886)||2012|
|Human||Not Cited||Src inhibitors in early breast cancer: a methodology, feasibility and variability study.||Jones RJ,Young O,Renshaw L,Jacobs V,Fennell M,Marshall A,Green TP,Elvin P,Womack C,Clack G,Dixon JM||Breast cancer research and treatment (114:211)||2009|
|Human||Not Cited||Mislocalization or reduced expression of Arf GTPase-activating protein ASAP1 inhibits cell spreading and migration by influencing Arf1 GTPase cycling.||Liu Y,Yerushalmi GM,Grigera PR,Parsons JT||The Journal of biological chemistry (280:8884)||2005|
|Mouse||Not Cited||Tyrosine phosphorylation of paxillin alpha is involved in temporospatial regulation of paxillin-containing focal adhesion formation and F-actin organization in motile cells.||Nakamura K,Yano H,Uchida H,Hashimoto S,Schaefer E,Sabe H||The Journal of biological chemistry (275:27155)||2000|
||Src inhibitors in early breast cancer: a methodology, feasibility and variability study.||Jones RJ,Young O,Renshaw L,Jacobs V,Fennell M,Marshall A,Green TP,Elvin P,Womack C,Clack G,Dixon JM||Breast cancer research and treatment (114:211)||2009|
|Human||Not Cited||Aberrant activation of focal adhesion proteins mediates fibrillar amyloid beta-induced neuronal dystrophy.||Grace EA,Busciglio J||The Journal of neuroscience : the official journal of the Society for Neuroscience (23:493)||2003|
||Tyrosine phosphorylation of paxillin alpha is involved in temporospatial regulation of paxillin-containing focal adhesion formation and F-actin organization in motile cells.||Nakamura K,Yano H,Uchida H,Hashimoto S,Schaefer E,Sabe H||The Journal of biological chemistry (275:27155)||2000|
PAX-1; PAXI; paxillin; paxillin alpha; testicular tissue protein Li 134
AW108311; AW123232; Pax; PXN