Immunofluorescence analysis of ETS [pS282/pS285] 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 ETS [pS282/pS285] Rabbit Polyclonal Antibody (441111G) 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 Nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Rat|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human ETS1 that contains both serine 282 and 285.|
|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 *|
|ChIP assay (ChIP)||5 µg|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Immunocytochemistry (ICC)||1 µg/ml|
|Immunofluorescence (IF)||1 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:50|
|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.
|Western Blot (WB)||See 1 publications below|
Ets-1 is a transcription factor involved in the regulation of genes required for extracellular matrix remodeling during metastasis of tumors. It is known to interact with the enhancer of urokinase like plasminogen activator, and promoters of stromelysin-1 and collagenase-1. It is absent from normal gastric mucosa but is expressed in 60% of gastric carcinoma and oral squamous cell carcinoma.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Phosphorylation of ETS1 by Src family kinases prevents its recognition by the COP1 tumor suppressor.
44-1111G was used in western blot to study how phosphorylation alters COP1-binding sites of oncoproteins.
|Lu G,Zhang Q,Huang Y,Song J,Tomaino R,Ehrenberger T,Lim E,Liu W,Bronson RT,Bowden M,Brock J,Krop IE,Dillon DA,Gygi SP,Mills GB,Richardson AL,Signoretti S,Yaffe MB,Kaelin WG||Cancer cell (26:222)||2014|