Western blot analysis of MET 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 MET (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. MET was detected at ~ 145 kDa using MET Mouse monoclonal antibody (Product # MA5-15198) diluted in Pierce Starting Block T20 (PBS) Blocking Buffer 4°C overnight on a rocking platform. Pierce Goat Anti-Mouse (product # 031437) 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). MET Antibody (Product # MA5-15198) confirms silencing of MET expression.
|Tested species reactivity||Mouse, Non-human primate, Rat, Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Synthetic peptide corresponding to residues surrounding pTyr1234 of human Met|
|Storage buffer||0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/ml BSA, 50% glycerol|
|Contains||<0.02% sodium azide|
|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.
The proto-oncogene MET product is the hepatocyte growth factor receptor and encodes tyrosine-kinase activity. The primary single chain precursor protein is post-translationally cleaved to produce the alpha and beta subunits, which are disulfide linked to form the mature receptor. Various mutations in the MET gene are associated with papillary renal carcinoma. Two transcript variants encoding different isoforms have been found for this gene.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.