Immunofluorescent analysis of MAPK1 (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were then blocked with 5% normal goat serum (Product #31873) for 15 minutes at room temperature. Cells were then probed with a mouse monoclonal antibody recognizing MAPK1 (Product# MA1-099), at a dilution of 1:200 for at least 1 hour at room temperature. Cells were then washed with PBS and incubated with DyLight 488 goat-anti-mouse secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product# 62249). Images were taken on a Thermo Scientific ArrayScan at 10X magnification.
|Tested species reactivity||Dog, Human, Mouse, Non-human primate, Rat|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Protein expressed in 293T cells transfected with human MAPK1 expression vector|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1-3 ul|
|Flow Cytometry (Flow)||1-10 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100-1:500|
|Immunoprecipitation (IP)||2 µg/500 µl|
|Western Blot (WB)||1-3 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.
MA1-099 has been successfully used in Western blot, ChIP, flow cytometry, immunofluorescence, immunohistochemistry and immunoprecipitation applications, and reacts with human, dog, rat, mouse and monkey samples.
MA1-099 has successfully been used in western blot to detect p42 ERK in HeLa, NRK and MDCK cell lysates.
As demonstrated by a Western blot using recombinant ERK1 and ERK2 proteins, MA1-099 can also detect ERK1, but detects ERK2 at much lower concentrations. As demonstrated by a Western blot using a panel of lysates from various species, MA1-099 only detects endogenous ERK2.
The protein encoded by this gene is a member of the MAP kinase family. MAP kinases, also known as extracellular signal-regulated kinases (ERKs), 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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