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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||Dog, Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human cofilin that contains serine 3. 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/10^6 cells|
|Immunocytochemistry (ICC)||2-3 ug/ml|
|Immunofluorescence (IF)||2-3 ug/ml|
|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.
Cofilin is a widely distributed intracellular actin-modulating protein that binds and depolymerizes filamentous F-actin and inhibits the polymerization of monomeric G-actin in a pH-dependent manner. It is involved in the translocation of actin-cofilin complex from cytoplasm to nucleus.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
18 kDa phosphoprotein; CFL; CFL-1; cofilin; Cofilin 1; cofilin 1 (non-muscle); Cofilin, non-muscle isoform; Cofilin-1; Cofilin1; epididymis secretory protein Li 15; non-muscle isoform; p18
AA959946; CFL; CFL1; Cof; cofilin; HEL-S-15; n-cofilin