Immunofluorescence analysis of Phospho-MEK1 pSer298 Antibody 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 Phospho-MEK1 pSer298Antibody (44460G) at 1:250 dilution in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa flour 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 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 594 Phalloidin (A12381). Panel d is a merged image showing cytoplasmic and nuclear localization. Panel e is a no primary antibody control. The images were captured at 40X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human MEK1 that contains serine 298. The sequence is conserved in many species including mouse, rat, chimp, hamster, and rabbit.|
|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)||1:20|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:200|
|Western Blot (WB)||1:500-1:2000|
* 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 2 publications below|
MAP2K1 is a dual specificity mitogen-activated protein kinase (MAPK) kinase. It is an essential component of the MAP kinase pathway and stimulates the enzymatic activity of MAP kinases in response to various signals. Growth factors, cytokines, and proto-oncogenes transduce their signals through the activation of RAS, which leads to activation of the serine/threonine kinase RAF, and then to activation of MAPK kinase. MAP2K1 is activated by the Raf family member p74raf-1, through phosphorylation of serine residues at positions 217 and 221 in the activation loop.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Group I Paks as therapeutic targets in NF2-deficient meningioma.
44-460G was used in western blot to study the molecular mechanisms underlying malignant changes of meningioma cells
|Chow HY,Dong B,Duron SG,Campbell DA,Ong CC,Hoeflich KP,Chang LS,Welling DB,Yang ZJ,Chernoff J||Oncotarget (6:1981)||2015|
|Human||Not Cited||The important roles of RET, VEGFR2 and the RAF/MEK/ERK pathway in cancer treatment with sorafenib.||Mao WF,Shao MH,Gao PT,Ma J,Li HJ,Li GL,Han BH,Yuan CG||Acta pharmacologica Sinica (33:1311)||2012|