Western blot analysis of SMAD1 was performed by loading 20 µg of C2C12 (lane1), HeLa (lane2) and Mouse kidney (lane3) lysates using Novex® NuPAGE® 4-12 % Bis-Tris gel (NP0321BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. SMAD1was detected at ~60 kDa using ABfinity™ SMAD1 Recombinant Rabbit Monoclonal Antibody (701168) at 1-3 µg/ml in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat Anti-Rabbit IgG-HRP Secondary Antibody (G21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (WP20005).
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide corresponding to amino acids 178–193 of human SMAD1|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||3µg|
|Flow Cytometry (Flow)||1-3µg/10^6 cells|
|Western Blot (WB)||1-3µg/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.
This antibody is predicted to react with equine, mouse, rabbit and rat based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
SMAD1 belongs to the SMAD family of proteins, which are downstream effectors of the TGF beta/BMP signaling pathway. Members of the SMAD family transmit TGF- beta signals from the cell surface into the nucleus. Once in the nucleus, SMADs can target a variety of DNA binding proteins to regulate transcriptional responses. SMAD1 is a receptor-regulated SMAD in the cell to regulate a wide range of critical processes including morphogenesis, cell-fate determination, proliferation, differentiation, and apoptosis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.