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|
|Published species reactivity||Not Applicable|
|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.
|Miscellaneous PubMed (MISC)||See 2 publications below|
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.
IP-MS enrichment of SMAD4 (LFQ intensity): SMAD4 was enriched 205-fold from HeLa lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and SMAD4 antibody (Part No. MA5-14300). The STRING database (www.string-db.org) was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Effect of DPC4 gene on invasion and metastasis of colorectal carcinoma cells.
MA5-14300 was used in immunohistochemistry and western blot to study the role of the DPC4 gene on invasion and metastasis of colorectal carcinoma cells
|Xiao DS,Wen JF,Li JH,Wang KS,Hu ZL,Zhou JH,Deng ZH,Liu Y||Acta biochimica et biophysica Sinica (38:883)||2006|
|Not Applicable||Not Cited||
Notch signaling modulates the nuclear localization of carboxy-terminal-phosphorylated smad2 and controls the competence of ectodermal cells for activin A.
MA5-14300 was used in immunoprecipitation and western blot to investigate the regulation of the onset of loss of mesodermal competence by Notch signaling during Xenopus development
|Abe T,Furue M,Kondow A,Matsuzaki K,Asashima M||Mechanisms of development (122:671)||2005|