Western blot analysis of MAP2K3 was performed with 10 µg of A549 cells transfected with Transfection Reagent alone (Lane 1), 100nM Non-Targeting control siRNA (Lane 2), or 100nM siRNA against MAP2K3 (Lane 3). Proteins were resolved using a NuPAGE® Novex 4-12% Bis-Tris Gel (NP0322BOX), XCell SureLock™ Electrophoresis System (EI0002), and a protein size ladder. Proteins were wet transferred to a Pierce Nitrocellulose Membrane (Product # 88025) OR Pierce PVDF Membrane (Product # 88518) and blocked with Pierce Starting Block T20 (PBS) Blocking Buffer (37539) for 1 hour at room temperature. MAP2K3 was detected at ~ 40 kDa using MAP2K3 Rabbit polyclonal antibody (Product # PA5-17480) diluted in Pierce Starting Block T20 (PBS) Blocking Buffer 4°C overnight on a rocking platform. Pierce Goat Anti-Rabbit (product # 31461) HRP-Conjugated Antibodies at a 1:2500 dilution were used and chemiluminescent detection was performed using Pierce Supersignal West Dura Maximum Sensitivity Substrate (Product # 37071). Relative density of the bands normalized to GAPDH (36 kDa). MAP2K3 Antibody (Product # PA5-17480) confirms silencing of MAP2K3 expression.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to the N-terminal sequence of human MKK3|
|Storage buffer||0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/ml BSA, 50% glycerol|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:1000|
* 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.
It is not recommended to aliquot this antibody.
This antibody is not cross-reactive with MEK1, MEK2, MKK4 or MKK6.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.