Anti-NOTCH2 Antibody, clone 1B14-5 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1830 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (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. ZRB1830, Anti-NOTCH2, clone 1B14-5 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in tubules and glomeruli of human kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (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. ZRB1830, Anti-NOTCH2, clone 1B14-5 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in tubules and glomeruli of human kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (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. ZRB1830, Anti-NOTCH2, clone 1B14-5 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in tubules and glomeruli of human kidney tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of MCF7 cells were performed using a 1:1,000 dilution of Cat. No. ZRB1830, Anti-NOTCH2, clone 1B14-5 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 MCF7 cells were performed using a 1:1,000 dilution of Cat. No. ZRB1830, Anti-NOTCH2, clone 1B14-5 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 MCF7 cells were performed using a 1:1,000 dilution of Cat. No. ZRB1830, Anti-NOTCH2, clone 1B14-5 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 NIH3T3 (Lane 1), C6 (Lane 2), and HeLa (Lane 3) cells were probed with Cat. No. ZRB1830, Anti-NOTCH2, clone 1B14-5 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 NOTCH2 (~110 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or Notch2 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1830, Anti-NOTCH2, clone 1B14-5 ZooMAb® Rabbit Monoclonal. The binding of this antibody to Notch2 peptide displayed a KD of 1.3 x 10-6, whereas binding to non-specific control peptide displayed a KD of 3.5 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 | 1B14, 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 ~265.41 kDa |
| observed mol wt ~110 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, mouse, rat |
| 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. | Q04721  |
| Application: | Quality Control Testing Evaluated by Western Blotting in HeLa cell lysate. Western Blotting Analysis: A 1:10,000 dilution of this antibody detected NOTCH2 in HeLa cell lysate. Tested Applications Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected NOTCH2 in NIH3T3 and C6 cell lysates. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected NOTCH2 in human kidney tissue sections. Affinity Binding Assay: A representative lot of this antibody bound Notch2 peptide with a KD of 1.3 x 10-6 in an affinity binding assay. Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected NOTCH2 in MCF7 cells. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-NOTCH2, clone 1B14-5 ZooMAb®, Cat. No. ZRB1830, is a recombinant Rabbit monoclonal antibody that specifically targets NOTCH2 and is tested for use in Affinity Binding Assay, Immunocytochemistry, Immunohistochemistry (Paraffin), and Western Blotting. |
| 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 18 amino acids from the C-terminal, cytoplasmic domain of human NOTCH2. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant mouse monoclonal antibody IgG, lyophilized in PBS, 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: | 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 1B14-5 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects NOTCH2. It targets an epitope within 18 amino acids from the C-terminal, cytoplasmic domain. |
| 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: | Neurogenic locus notch homolog protein 2 (UniProt: Q04721; also known as Notch 2, hN2, NOTCH2) is encoded by the NOTCH2 gene (Gene ID: 4853) in human. Notch is a single-pass type I membrane protein that is synthesized with a signal peptide (aa 1-25), which is subsequently cleaved off to generate the mature form that contains a long extracellular domain (aa 26-2677), a transmembrane domain (aa 1678-1698), and a cytoplasmic domain (aa 1699-2471). It is evolutionarily conserved and is implicated in cell-fate determination, tissue patterning, morphogenesis, and cell differentiation. It is expressed in the brain, heart, kidney, lung, skeletal muscle and liver and is ubiquitously expressed in the embryo. Notch is synthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved by a furin-like convertase in the trans-Golgi network before it reaches the plasma membrane to yield an active, ligand-accessible form. It serves as a receptor for Jagged-1, Jagged-2, and Delta-1 to regulate cell-fate determination. Following ligand binding, Notch is cleaved by TNF-alpha converting enzyme (S2 cleavage) to yield a membrane-associated intermediate fragment called notch extracellular truncation, which is then cleaved by g-secretase (S3 cleavage) to release a notch-derived peptide containing the intracellular domain (NICD; aa 1697-2471) from the membrane that translocates to the nucleus and initiates transcription events by binding the DNA binding protein CSL. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Yin, L., et al (2010). Biochem. Pharmacol. 80(5); 690-701). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |