Immunofluorescence analysis of MEK1 / MAP2K1 was performed using 70% confluent log phase HeLa cells. 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 MEK1 / MAP2K1 Rabbit Polyclonal Antibody (PA5-16556) at 2 µg/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 (A27034) 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 represents the merged image showing punctated cytoplasmic and nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat, Xenopus|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide derived from the N-terminus of human Mek1 protein|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Western Blot (WB)||0.5-2 µg/ml|
* 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.
PA5-16556 targets MEK-1 in WB applications and shows reactivity with mouse, Rat, Xn, and Human samples.
The PA5-16556 immunogen is a synthetic peptide derived from the N-terminus of human Mek1 protein.
Mek1/mitogen activated protein kinase kinase/ERK kinase 1/microtubule-associated protein 2 kinase is a dual specificity kinase and catalyses both a threonine and a tyrosine residue on MAP kinases ERK1 and ERK2. Mek1 is activated by phosphorylation of serine 218 and 222 residues by Raf1. It is known to be involved in the signaling during stress activate response, apoptosis and proliferative induction by cytokines.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.