Anti-phospho-IRS1 (Tyr612) Antibody, clone 1N14, ZooMAb® Rabbit Monoclonal
SIGMA/ZRB09432 - recombinant, expressed in HEK 293 cells
Synonym: IRS-1; Insulin Receptor Substrate 1
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human breast cancer (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB09432, Anti-phospho-IRS1 (Tyr612), clone 1N14, ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/nuclear staining was observed in human breast cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human breast cancer (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB09432, Anti-phospho-IRS1 (Tyr612), clone 1N14, ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/nuclear staining was observed in human breast cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human breast cancer (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB09432, Anti-phospho-IRS1 (Tyr612), clone 1N14, ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/nuclear staining was observed in human breast cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human prostate cancer (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB09432, Anti-phospho-IRS1 (Tyr612), clone 1N14, ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/nuclear staining was observed in human prostate cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human prostate cancer (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB09432, Anti-phospho-IRS1 (Tyr612), clone 1N14, ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/nuclear staining was observed in human prostate cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human prostate cancer (A) and Negative control (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB09432, Anti-phospho-IRS1 (Tyr612), clone 1N14, ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/nuclear staining was observed in human prostate cancer tissue sections.
Western Blotting Enhanced Validation-Recombinant Antibody Technology Lysates from insulin treated HEK293 (A) and NIH3T3 (B) cells were probed with Cat. No. ZRB09432, Anti-phospho-IRS1 (Tyr612), clone 1N14, 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-IRS1 (Tyr612) (~160 kDa).
Peptide Inhibition Enhanced Validation-Recombinant Antibody Technology Lysates from Insulin treated HEK293 cells were probed with Cat. No. ZRB09432, Anti-phospho-IRS1 (Tyr612), clone 1N14, ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminesence detection system. Lane 1: No peptide Lane 2: With phospho IRS1 (Tyr612) peptide Lane 3: With non-phospho IRS1 peptide Arrow indicates phospho-IRS1 (Tyr612) ( ~160 kDa).
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 | 1N14, 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 131.59 kDa |
| observed mol wt ~160 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | mouse, human |
| storage temp. | 2-8°C |
| technique(s) | immunohistochemistry: suitable using 1:100 |
| inhibition assay: suitable using 1:1,000 (peptide) | |
| western blot: suitable using 1:1,000 | |
| UniProt accession no. | P35568  |
| Application: | Anti-phospho-IRS1 (Tyr612), clone 1N14, ZooMAb is a recombinant Rabbit Monoclonal antibody that detects Insulin receptor substrate 1 (IRS1) and has been tested for use in Immunohistochemistry, Peptide Inhibition Assay, and Western Blotting. |
| Application: | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected phospho-IRS1 (Tyr612) in lysate from NIH3T3 cells treated with Insulin. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Insulin receptor substrate 1 in human breast cancer and human prostate cancer tissue sections. Peptide Inhibition Analysis: A 1:1,000 dilution from a representative lot was used with insulin treated HEK293 cells for phosphopeptide block analysis. |
| 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 11 amino acids surrounding phospho tyrosine 612 of human Insulin receptor substrate 1 (IRS-1). |
| 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: | 30 μg/mL after reconstitution at 25 μL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 1N14 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects human IRS-1 phosphorylated on Tyrosine 612 and murine IRS-1 phosphorylated on Tyrosine 608. |
| 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: | Insulin receptor substrate 1 (UniProt: P35568; also known as IRS-1) is encoded by the IRS1 gene (Gene ID: 3667) in human. The insulin receptor substrates (IRS-1, IRS-2, IRS-3, and IRS-4) are adaptor proteins involved in modulating cell growth, metabolism, survival, and differentiation by transducing receptor signaling to multiple downstream effectors. IRS family proteins contain a conserved N-terminal pleckstrin homology (PH) domain (aa 12-115 of human IRS-1), a phosphotyrosine-binding (PTB) domain (aa 160-264 of human IRS-1), and multiple tyrosine phosphorylation sites in the C-terminal region. The PTB domain mediates the interaction of IRS-1, -2 and -3 with IR juxtamembrane NPXYp motif, facilitating subsequent receptor-mediated IRS tyrosine phosphorylation. Upon phosphorylation, IRS proteins serve as binding sites for Src homology 2 (SH2) domain-containing proteins, such as SHP-2 and PI 3-kinase. Functions of IRS proteins are largely dependent on the phosphorylation of specific tyrosine residues, which in turn are subject to negative regulation by protein phosphatase activity. In addition, IRS proteins can be phosphorylated by serine/threonine kinases, such as Akt/PKB, GSK-3, mTOR, and PKC, and increased Ser/Thr phosphorylation reduces their binding to IR and tyrosine phosphorylation. When phosphorylated, tyrosine 612 is a pYMxM consensus motif PI3-kinase binding site, which leads to the activation of the PI3K/AKT signaling pathway. Inhibition of PI 3-kinase enhances insulin-stimulated IRS-1 Tyr612/Tyr941 phosphorylation (p85-binding sites) and increases IRS-1 association with PI 3-kinase p85 subunit. Defects in muscle IRS1 gene expression and function have been reported in insulin-resistant states, such as in obesity and type 2 diabetes. 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 |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |