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Immunofluorescence analysis of Phospho-Paxillin pSer126 was done on 70% confluent log phase NIH/3T3 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-Paxillin pSer126 Rabbit Polyclonal Antibody (441022G) 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 punctuated nuclear 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||Chicken, 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 serine 126. The sequence is conserved in mouse and chicken.|
|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 *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2-3 µg/mL|
|Immunofluorescence (IF)||2-3 µg/mL|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Western Blot (WB)||1:1000|
* 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.
PAX-1; PAXI; paxillin; paxillin alpha; testicular tissue protein Li 134
AW108311; AW123232; Pax; PXN