Selective non-competitive inhibitor of MAP kinase kinase (MKK). Inhibits MKK1, MKK2 (IC50 values of 0.07 and 0.06 μM, respectively) and MKK5 with little or no effect on a wide range of other kinases. Centrally active following systemic administration in vivo.
Store at +4°C. Store under desiccating conditions. The product can be stored for up to 12 months.
Soluble in DMSO to 100 mM
Wherever possible, you should prepare and use solutions on the same day. However, if you need to make up stock solutions in advance, we recommend that you store the solution as aliquots in tightly sealed vials at -20°C. Generally, these will be useable for up to one month. Before use, and prior to opening the vial we recommend that you allow your product to equilibrate to room temperature for at least 1 hour.
Performed under reducing conditions; Samples loaded at 30 µg/lane. Membrane blocking and secondary antibody incubation performed in 5% milk/PBST. Primary antibody incubation performed in 5% BSA/PBST.
Lane 1: Marker
Lane 2: A431 lysate starved
Lane 3: A431 lysate starved treated with 100 ng/mL of EGF for 30 min
Lane 4: A431 lysate starved and pretreated for 2 hours with 10µM U0126 (ab120241)
Lane 5: A431 lysate starved and pretreated for 2 hours with 30µM Ly294002 (ab120243)
Primary: AKT/MAPK signaling pathway antibody cocktail at 1/250 dilution
Secondary: HRP conjugated Goat anti-Rabbit secondary antibody at 1:10,000 dilution.
RSK1p90 (pS380) band size: 83 kDa
AKT1 (pS473) band size: 56 kDa
ERK1 (pY204) and ERK2 (pY187) band size: 43 kDa
RPS6 (pS235/236) band size: 31 kDa
Rab11a band size: 24 kDa
Unknown band: 130kDa