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Immunofluorescence analysis of Paxillin was done on 70% confluent log phase HeLa cells. 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 Paxillin (5H11) Mouse Monoclonal Antibody (AHO0492) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) 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 punctuated cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Dog, Rat, Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Purified recombinant full-length human paxillin protein.|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunofluorescence (IF)||Assay Dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||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.
Staining of formalin-fixed, paraffin-embedded tissues requires boiling tissues in 1mM EDTA, pH 8.0, for 10-20 min followed by cooling at room temperature for 20 min. Recommended positive controls include HeLa cells and colon carcinoma.
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.
αV-integrins are required for mechanotransduction in MDCK epithelial cells.
AHO0492 was used in immunocytochemistry to demonstrate that αV-integrins regulate the maturation of focal adhesions and cell spreading.
|Teräväinen TP,Myllymäki SM,Friedrichs J,Strohmeyer N,Moyano JV,Wu C,Matlin KS,Muller DJ,Manninen A||PloS one (8:null)||2013|
Paxillin mutations affect focal adhesions and lead to altered mitochondrial dynamics: relevance to lung cancer.
AHO0492 was used in immunocytochemistry and western blot to examine the effects of activating mutations of PXN on mitochondrial function.
|Kawada I,Hasina R,Lennon FE,Bindokas VP,Usatyuk P,Tan YH,Krishnaswamy S,Arif Q,Carey G,Hseu RD,Robinson M,Tretiakova M,Brand TM,Iida M,Ferguson MK,Wheeler DL,Husain AN,Natarajan V,Vokes EE,Singleton PA,Salgia R||Cancer biology & therapy (14:679)||2013|
c-Yes regulates cell adhesion at the blood-testis barrier and the apical ectoplasmic specialization in the seminiferous epithelium of rat testes.
AHO0492 was used in western blot to elucidate how c-Yes regulates the blood-testis barrier and apical ectoplasmic specialization integrity.
|Xiao X,Mruk DD,Lee WM,Cheng CY||The international journal of biochemistry & cell biology (43:651)||2011|
|Human||Not Cited||Loss of caveolin-1 polarity impedes endothelial cell polarization and directional movement.||Beardsley A,Fang K,Mertz H,Castranova V,Friend S,Liu J||The Journal of biological chemistry (280:3541)||2005|
|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|
myocardial ischemic preconditioning associated protein 7; PAX-1; PAXI; paxillin; paxillin alpha; testicular tissue protein Li 134
AW108311; AW123232; Pax; PXN