Anti-SMAD4 Antibody, clone 1C14 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1981 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from HepG2 (Lane 1), A431 (Lane 2), and NIH3T3 (Lane 3) cells were probed with Cat. No. ZRB1981, Anti-SMAD4, clone 1C14 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates SMAD4 (~59 kDa).
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from wild-type HeLa cells (Lane 1) and SMAD4 Knockout HeLa cells (Lane 2) were probed with Cat. No. ZRB1981, Anti-SMAD4, clone 1C14 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates SMAD4 (~59 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of SMAD4 peptide (Run 1) or non-specific control peptide (Run 2) or were analyzed by affinity binding assay using Cat. No. ZRB1981, Anti-SMAD4, clone 1C14 ZooMAb® Rabbit Monoclonal. The binding of this antibody to SMAD4 peptide displayed a KD of 1.0 x 10-12 whereas binding to non-specific control peptide displayed a KD of 7.8 x 10-4.
ELISA Enhanced Validation - Recombinant Antibody Technology 100 mL of 1 mg/mL of SMAD4 peptide (Blue) and control peptide (Orange) were analyzed by ELISA using 3,000 ng/mL to 1 ng/mL dilutions of Cat. No. ZRB1981, Anti-SMAD4, clone 1C14 ZooMAb® Rabbit Monoclonal. Alkaline phosphatase-conjugated Goat Anti-Rabbit IgG Fc was used for detection.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 1C14, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | C-terminus |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 60.44 kDa |
| observed mol wt ~59 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_005350.1?  |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human, mouse |
| species reactivity (predicted by homology) | bovine, monkey |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| ELISA: suitable | |
| western blot: suitable | |
| UniProt accession no. | Q13485 |
| Application: | Quality Control Testing Evaluated by Western Blotting in NIH3T3 cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected SMAD4 in NIH3T3 cell lysate. Tested applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected SMAD4 in HepG2 and A431 cell lysates. Western Blotting KO Analysis: A 1:1,000 dilution from a representative lot detected SMAD4 in lysate from wild-type HeLa cells but not in SMAD4 knockout HeLa lysate. Affinity Binding Assay: A representative lot of this antibody bound SMAD4 with a KD of 1.0 x 10-12 in an affinity binding assay. ELISA: A representative lot of this antibody detected SMAD4 in ELISA application. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-SMAD4, clone 1C14 ZooMAb®, Cat. No. ZRB1981, is a recombinant Rabbit monoclonal antibody that specifically targets SMAD4 and is tested for use in Affinity Binding Assay, ELISA, Western Blotting, and Western Blotting using Knockout lysates. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them. Learn more about ZooMAb here. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 16 amino acids from the C-terminal region of human SMAD4. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 1C14 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects SMAD4. It targets an epitope within 16 amino acids from the C-terminal region. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Mothers against decapentaplegic homolog 4 (UniProt: Q13485; also known as MAD homolog 4, Mothers against DPP homolog 4, Deletion target in pancreatic carcinoma 4, SMAD family member 4, SMAD 4, Smad4, hSMAD4) is encoded by the SMAD4 (also known as DPC4, MADH4) gene (Gene ID: 4089) in human. SMAD proteins are signal transducers for the members of the transforming growth factor-b (TGF-b) superfamily. SMAD4 is a transcription factor that is involved in TGF-β signaling. In the absence of a ligand it is monomeric and is localized in the cytoplasm. Upon ligand binding it forms a heterotrimer with SMAD2 or SMAD3 and another molecule of SMAD4 to form the active complex that translocates to the nucleus. SMAD4 can undergo monoubiquitination on lysine 519 by E3 ubiquitin-protein ligase TRIM33 and this monoubiquitination hampers its ability to form a stable complex with activated SMAD2/3 resulting in inhibition of TGF-b signaling. Deubiquitination by USP9X is reported to restore its competence to mediate TGF-b signaling. As the main effector of TGF-β signaling, SMAD4 has been found to be non-functional in more than half of adenocarcinomas of the pancreatic duct and in colorectal cancers. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Chen, HB., et al. (2005). J. Biol. Chem. 280(22); 21329-21936; Kawabata, M., et al. (1998). EMBO J. 17(14); 4056-4065). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |