Anti-phospho-Stat1-Tyr701 Antibody, clone 1F23 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1520 - recombinant, expressed in HEK 293 cells
Synonym: Phospho-STAT1 (Tyr701); Signal transducer and activator of transcription 1; Transcription factor ISGF-3 components p91/p84
Product Type: Chemical
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells, serum starved and treated with interferon a (2000 units/mL; 20 min.) was performed using a 1:100 dilution of Cat. No. ZRB1520, Anti-phospho-Stat1-Tyr701, clone 1F23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells, serum starved and treated with interferon a (2000 units/mL; 20 min.) was performed using a 1:100 dilution of Cat. No. ZRB1520, Anti-phospho-Stat1-Tyr701, clone 1F23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells, serum starved and treated with interferon a (2000 units/mL; 20 min.) was performed using a 1:100 dilution of Cat. No. ZRB1520, Anti-phospho-Stat1-Tyr701, clone 1F23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488. Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Western Blotting Enhanced Validation-Recombinant Antibody Technology Lysates from HeLa cells (Lane 1) and HeLa cells stimulated with interferon a (2,000 units/mL; 20 min.) (Lane 2) were probed with Cat. No. ZRB1520, Anti-phospho-Stat1-Tyr701, clone 1F23 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 phospho-Stat1-Tyr701 (~91 kDa).
Affinity Binding Assay Enhanced Validation-Recombinant Antibody Technology 100 μg/mL of phosphorylated Stat1-Tyr701 peptide (Run 1) non-phosphorylated Stat1 control peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1520, Anti-phospho-Stat1-Tyr701, clone 1F23 ZooMAb® Rabbit Monoclonal. The binding of this antibody to phosphorylated Stat1-Tyr701 peptide displayed a KD value of 7.9 x 106 whereas its binding to non-phosphorylated Stat1 control peptide displayed a KD of 2.4 x 103.
Flow Cytometry Enhanced Validation-Recombinant Antibody Technology Staining of one million HeLa cells stimulated with IFN-alpha (2,000 units/mL; 20 min.) was performed using 1 μg of Cat. No. ZRB1520, Anti-phospho-Stat1-Tyr701, clone 1F23 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.
Peptide Inhibition Assay Enhanced Validation-Recombinant Antibody Technology Lysates from HeLa cells stimulated with Interferon a (2,000 units/mL; 20 min) were subjected to immunogen peptide exposures and probed with Cat. No. ZRB1520, Anti-phospho-Stat1-Tyr701, clone 1F23 ZooMAb® Rabbit Monoclonal (1:500 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-Stat1-Tyr701 (~91 kDa). Lane 1: Without treatment with immunogen peptide Lane 2: Treated with phosphorylated immunogen peptide Lane 3: Treated with non-phosphorylated peptide
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 (recombinant) |
| clone | 1F23, monoclonal |
| recombinant monoclonal | |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| 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 87.33 kDa |
| observed mol wt ~91 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| species reactivity (predicted by homology) | bovine, monkey |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| flow cytometry: suitable | |
| immunocytochemistry: suitable | |
| inhibition assay: suitable | |
| western blot: suitable | |
| UniProt accession no. | P42224  |
| Application: | Anti-p-Stat1-Tyr701, clone 1F23 ZooMAb, Cat. No. ZRB1520, is a recombinant Rabbit monoclonal antibody that targets p-Stat1 (Y701) and is used in Affinity Binding Assay, Flow Cytometry, Immunocytochemistry, Peptide Inhibition Assay, and Western Blotting. |
| Application: | Flow Cytometry Analysis: 1 μg from a representative lot detected phospho-Stat1-Tyr701 in HeLa cells treated with interferon a (2000 Units/mL; 20 min.) Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected phospho-Stat1-Tyr701 in HeLa cells, serum starved and treated with interferon a (2000 Units/mL; 20 min.). Peptide Inhibition Assay: Target band detection in 10 μg of lysate from HeLa cells treated with interferon a was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide. Affinity Binding Assay: A representative lot of this antibody bound phosphorylated Stat1-Tyr701 peptide with a KD of 7.9 x 10-6 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. Learn more about ZooMAb here. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 12 amino acids surrounding phosphotyrosine 701 from the C-terminal region of human Signal transducer and activator of transcription 1 (Stat1). |
| 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. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 0.3 mg/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 1F23 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects Stat1 phosphorylated on Tyrosine 701. It targets an epitope within 12 amino acids surrounding phosphotyrosine 701 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 1 (UniProt: P42224, also known as Transcription factor ISGF-3 components p91/p84) is encoded by the STAT1 gene (Gene ID: 6772) in human. STATs (signal transducers and activators of transcription) are a family of cytoplasmic latent transcription factors that are activated to regulate gene expression in response to a large number of extracellular signaling proteins. STAT1 is activated by many different ligands including interferons, interleukins, and growth factors. Following ligand binding to cell surface receptors, signaling via protein kinases leads to activation of JAK kinases and tyrosine phosphorylation of STAT1 and STAT2. The phosphorylated STATs dimerize and enter the nucleus where it activates expression of multiple proinflammatory genes, including inducible nitric oxide synthase (iNOS) and cyclooxygenase. Loss of STAT1 activity can lead to early fatal infections and increased susceptibility to severe mycobacterial and viral infections. Phosphorylation of tyrosine 701 is required for STAT 1 dissociation from IFNGR1, homodimerization, and nuclear translocation. Tyrosine 701 phosphorylation impairment results in loss of STAT 1 functions. Phosphorylation on serine 727 by several kinases including MAPK14, ERK1/2 and CAMKII following on IFN-gamma stimulation, regulates STAT1 transcriptional activity. This phosphorylation also promotes sumoylation though increasing interaction with PIAS. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |