Immunofluorescence analysis of Phospho- MEK1[pSpS 218/222]/MEK2 [pSpS222/226] 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[pSpS 218/222]/MEK2 [pSpS222/226] Antibody (44454G) at 1µg/mL 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 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||Synthetic phosphopeptide derived from a region of human MEK1 that contains serine 218 and serine 222|
|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/million cells|
|Western Blot (WB)||1:500-1:2000|
|Western Blot (WB)||See 1 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.
|Human||Not Cited||Toll-like receptor-mediated production of IL-1Ra is negatively regulated by GSK3 via the MAPK ERK1/2.||Rehani K,Wang H,Garcia CA,Kinane DF,Martin M||Journal of immunology (Baltimore, Md. : 1950) (182:547)||2009|
dual specificity mitogen-activated protein kinase kinase 1; dual specificity mitogen-activated protein kinase kinase 2; EC 220.127.116.11; ERK activator kinase 1; ERK activator kinase 2; kinase MEK1; MAP kinase kinase 1; MAP kinase kinase 2; MAP kinase/Erk kinase; MAP kinase/Erk kinase 1; MAP2K1; MAPK/ERK kinase 1; MAPK/ERK kinase 2; MAPKK 1; MAPKK 2; MAPKK1; MEK 1; MEK 2; MEK1; mitogen activated protein kinase kinase 1; mitogen activated protein kinase kinase 2; mitogen-activated protein kinase kinase 1; mitogen-activated protein kinase kinase 2, p45; MP2K1; PRKMK1; protein kinase, mitogen activated, kinase 1, p45; protein kinase, mitogen activated, kinase 2, p45; protein kinase, mitogen-activated, kinase 1 (MAP kinase kinase 1)
AA589381; CFC3; CFC4; MAP2K1; MAP2K2; MAPKK1; MAPKK2; MEK1; MEK2; MEKK1; MK2; MKK1; MKK2; PRKMK1; PRKMK2