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Western blot analysis of SMAD4 was performed with 10 µg of HeLa cells transfected with Transfection Reagent alone (Lane 1), 100nM Non-Targeting control siRNA (Lane 2), or 100nM siRNA against SMAD4 (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. SMAD4 was detected at ~ 60 kDa using SMAD4 Mouse monoclonal antibody (Product # MA5-14300) 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 Actin (45 kDa). SMAD4 Antibody (Product # MA5-14300) confirms silencing of SMAD4 expression.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified recombinant DPC4 protein|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||Assay Dependent|
|Immunoprecipitation (IP)||2 µg/ml|
|Western Blot (WB)||1-2 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.
MA5-14300 targets SMAD4 in IF, IP, and WB applications and shows reactivity with Human samples.
The MA5-14300 immunogen is purified recombinant DPC4 protein.
DPC4 (also named SMAD4; small mothers against decapentaplegic deleted in pancreatic carcinoma). The family of SMAD molecules comprises essential mediators of transforming growth factor beta (TGF-beta) signaling. To date, seven members of this family have been identified, each of which plays a specific and separate role in mediating TGF-beta superfamily gene transcription. At least two different members, Smad2 and Smad4/DPC4, have been implicated in human cancer and appear to have tumor-suppressor functions whose inactivation may play an important role in the histogenesis of pancreatic and colorectal malignancy and possibly other human cancers as well.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
deleted in pancreatic carcinoma locus 4; deletion target in pancreatic carcinoma 4; Drosophila; homolog of; hSMAD4; MAD; SMAD; 4; MAD homolog 4; mothers against decapentaplegic; mothers against decapentaplegic homolog 4; mothers against decapentaplegic, Drosophila, homolog of, 4; mothers against DPP homolog 4; SMAD 4; SMAD, mothers against DPP homolog 4
DPC4; JIP; MADH4; MYHRS; SMAD4