Anti-Stat3 Antibody, clone 2D2 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1764 - recombinant, expressed in HEK 293 cells
Synonym: Signal transducer and activator of transcription 3;Acute-phase response factor
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Lung cancer (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. ZRB1764, Anti-Stat3, clone 2D2 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in Human Lung cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Lung cancer (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. ZRB1764, Anti-Stat3, clone 2D2 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in Human Lung cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Lung cancer (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. ZRB1764, Anti-Stat3, clone 2D2 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in Human Lung cancer tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1764, Anti-Stat3, clone 2D2 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 cytoplasm and nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1764, Anti-Stat3, clone 2D2 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 cytoplasm and nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1764, Anti-Stat3, clone 2D2 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 cytoplasm and nucleus.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from Cos7 (Lane 1), A549 (Lane 2), Jurkat (Lane 3), C6 (Lane 4), and HepG2 (Lane 5) cells were probed with Cat. No. ZRB1764, Anti-Stat3, clone 2D2 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Stat3 (~90 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or Stat3 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1764, Anti-Stat3, clone 2D2 ZooMAb® Rabbit Monoclonal. The binding of this antibody to Stat3 peptide displayed a KD of 3.8 x 10-8, whereas binding to non-specific control peptide displayed a KD of 4.0 x 10-3.
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million Jurkat cells was performed using 0.1 μg of Cat. No. ZRB1764, Anti-Stat3, clone 2D2 ZooMAb® Rabbit Monoclonal (Yellow histogram) or the equivalent amount of a Rabbit IgG isotype control (Grey histogram), followed by a PE-conjugated Donkey Anti-Rabbit IgG secondary antibody.
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 | 2D2, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | C-terminal |
| 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 88.07 kDa |
| observed mol wt ~90 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_644805  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | rat, human, monkey |
| species reactivity (predicted by homology) | feline, mouse, porcine, bovine, canine |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| flow cytometry: suitable | |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | P40763 |
| Application: | Quality Control Testing Evaluated by Western Blotting in C6 cell lysate. Western Blotting Analysis: A 1:10,000 dilution of this antibody detected Stat3 in C6 cell lysate. Tested applications Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected Stat3 in Cos7, A549, Jurkat, and HepG2 cell lysates. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Stat3 in Jurkat cells. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Stat3 in human lung cancer tissue sections. Flow Cytometry Analysis: 0.1 μg from a representative lot detected Stat3 in one million Jurkat cells. Affinity Binding Assay: A representative lot of this antibody bound Stat3 with a KD of 3.8 x 10-8 in an affinity binding assay. 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 16 amino acids from the C-terminal region of human Stat3. |
| 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 2D2 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects Stat3. 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: | Signal transducer and activator of transcription 3 (UniProt: P40763; also known as Stat3, Acute-phase response factor) is encoded by the STAT3 (also known as APRF) gene (Gene ID: 6774) in human. STAT3 is expressed in most cell types and is predominantly cytoplasmic in its latent form. It serves as a cytoplasmic transcription factor that has a dual function of signal transduction and activation of transcription. It is activated by multiple cytokines, growth factors, and leptin. Stat3 is shown to shuttle between the nucleus and the cytoplasm. It is activated by phosphorylation at Tyrosine 705, which induces dimerization, nuclear translocation, and DNA binding. A motif comprising of amino acids 150-162 is essential for its nuclear import. In the nucleus it recruits coactivators, such as NCOA1 or MED1, to the promoter region of the target genes. It binds to DNA as either a homodimer or as a heterodimer with Stat1. Stat3 regulates inflammatory response by regulating differentiation of naive CD4+ T-cells into T-helper Th17 or regulatory T-cells. It may also play an anti-apoptotic role by transactivating BIRC5 expression under leptin activation. Cytoplasmic Stat3 is shown to repress macroautophagy by inhibiting EIF2AK2/PKR activity. Stat3 is also involved in cell cycle regulation by inducing the expression of key genes that are involved in the progression from G1 to S phase. Constitutively activated Stat3 is reported in a number of human cancers. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Shen, S., et al. (2012). Mol. Cell. 48(5); 667-680; Jiang, H., et al. (2008). Biochem. Biophys. Res. Commun. 368(1); 1-5; Levy, DE., and Lee C-K. (2002). J. Clin. Invest. 109(9); 1143-1148). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |