Anti-Smad2 Antibody, clone 2F15 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1765 - recombinant, expressed in HEK 293 cells
Synonym: Mothers against decapentaplegic homolog 2;MAD homolog 2;Mothers against DPP homolog 2;JV18-1;Mad-related protein 2;hMAD-2;SMAD family member 2;SMAD 2;hSMAD2
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Testis (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1765, Anti-Smad2, clone 2F15 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in seminiferous tubules of Human Testis tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Testis (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1765, Anti-Smad2, clone 2F15 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in seminiferous tubules of Human Testis tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Testis (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1765, Anti-Smad2, clone 2F15 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in seminiferous tubules of Human Testis tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of MCF-7 cells was performed using a 1:100 dilution of Cat. No. ZRB1765, Anti-Smad2, clone 2F15 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus and cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of MCF-7 cells was performed using a 1:100 dilution of Cat. No. ZRB1765, Anti-Smad2, clone 2F15 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus and cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of MCF-7 cells was performed using a 1:100 dilution of Cat. No. ZRB1765, Anti-Smad2, clone 2F15 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus and cytoplasm.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from HT1080 (Lane 1), C6 (Lane 2), HT29 (Lane 3), and MCF7 (Lane 4) cells were probed with Cat. No. ZRB1765, Anti-Smad2, clone 2F15 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 Smad2 (~55 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or Smad2 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1765, Anti-Smad2, clone 2F15 ZooMAb® Rabbit Monoclonal. The binding of this antibody to Smad2 peptide displayed a KD of 4.4 x 10-8, whereas binding to non-specific control peptide displayed a KD of 2.0 x 10-3.
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 | 2F15, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | N-terminal half |
| 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 52.31 kDa |
| observed mol wt ~55 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_001003652  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human, rat |
| species reactivity (predicted by homology) | monkey, bovine, porcine |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | Q15796 |
| Application: | Quality Control Testing Evaluated by Western Blotting in MCF7 cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Smad2 in MCF7 cell lysate. Tested applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected Smad2 in HT1080, C6, and HT29 cell lysates. Affinity Binding Assay: A representative lot of this antibody bound Smad2 with a KD of 4.4 x 10-8 in an affinity binding assay. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Smad2 in MCF-7 cells. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Smad2 in human testis tissue sections. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| 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. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 24 amino acids from the N-terminal half of human Smad2. |
| 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 2F15 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects Smad2. It targets an epitope within 24 amino acids from the N-terminal half. |
| 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 2 (UniProt: Q15796; also known as MAD homolog 2, Mothers against DPP homolog 2, JV18-1, Mad-related protein 2, hMAD-2, SMAD family member 2, SMAD 2, Smad2, hSMAD2) is encoded by the SMAD2 (also known as MADH2, MADR2) gene (Gene ID: 4087) in human. Smad2 is a receptor-regulated Smad (R-Smad) that serves as an intracellular signal transducer and transcriptional modulator activated by transforming growth factor-b (TGF-b) and activin type 1 receptor kinases. In the absence of TGF-b, it exists as a monomer and forms a heterodimer with Smad4 in the presence of TGF-b. It is expressed at high levels in skeletal muscle, endothelial cells, heart, and placenta. It contains one Mad homology (MH) 1 domain (aa 10-176), one MH2 domain (aa 274-467) and a PY motif (aa 221-225). Cytoplasmic Smad2 is directly phosphorylated on Ser465/Ser467 by TGF-b receptor kinase I. These C-terminal phosphoserines of R-Smads are recognized by the MH2 domain of Smad4 that enables them to form a heterodimeric complex, which translocates to the nucleus where Smad proteins bind to their cognate DNA binding sites with low affinity. This binding is further enhanced in the presence of transcriptional co-activators. Upon dephosphorylation by phosphatase PPM1A, Smad2 is released from the Smad2/Smad4 complex and is exported out of the nucleus. Acetylation of lysine 19 by coactivators in response to TGF- signaling is shown to increase its transcriptional activity. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Lin, X., et al. (2006). Cell. 125(5); 915-928; Simonsson, M., et al. (2006). J. Biol. Chem. 281(52); 39870-39880). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |