Immunofluorescent analysis of p44,42 MAPK (Erk1/2) in HeLa cells. The cells were fixed with 4% paraformaldehyde in PBS for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 15 minutes, and blocked with 3% BSA for 30 minutes at room temperature. Cells were stained with a p44/42 MAPK (Erk1/2) mouse monoclonal antibody (Product # MA5-15605) at a dilution of 1:250 in blocking buffer for 1 hour at room temperature, and then incubated with a Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:1000 for at least 30 minutes at a room temperature in the dark (green). Nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Purified recombinant fragment of human MAPK expressed in E. Coli.|
|Storage buffer||PBS with 1mg/ml BSA|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1/200 - 1/400|
|Immunohistochemistry (IHC)||1/200 - 1/1000|
|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.
MA5-15605 targets p44/42 MAPK (Erk1/2) in FACS, IHC, WB and IF applications and shows reactivity with Human and mouse samples.
The protein encoded by this gene is a member of the MAP kinase family. MAP kinases, also known as extracellular signal-regulated kinases , act as an integration point for multiple biochemical signals, and are involved in a wide variety of cellular processes such as proliferation, differentiation, transcription regulation and development. The activation of this kinase requires its phosphorylation by upstream kinases. Upon activation, this kinase translocates to the nucleus of the stimulated cells, where it phosphorylates nuclear targets. Two alternatively spliced transcript variants encoding the same protein, but differing in the UTRs, have been reported for this gene.
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