Anti-Notch1 Antibody, clone 1A32 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1943 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:200 dilution of Cat. No. ZRB1943, Anti-Notch1, clone 1A32 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.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:200 dilution of Cat. No. ZRB1943, Anti-Notch1, clone 1A32 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.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:200 dilution of Cat. No. ZRB1943, Anti-Notch1, clone 1A32 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.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:200 dilution of Cat. No. ZRB1943, Anti-Notch1, clone 1A32 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.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:200 dilution of Cat. No. ZRB1943, Anti-Notch1, clone 1A32 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.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:200 dilution of Cat. No. ZRB1943, Anti-Notch1, clone 1A32 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.
Western Blotting Enhanced Validation - Recombinant Antibody Technology HEK293 cell lysate was probed with Cat. No. ZRB1943, Anti-Notch1, clone 1A32 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 Notch1 (~125 kDa).
Western Blotting Enhanced Validation - Recombinant Antibody Technology A 1:1,000 dilution of Cat. No. ZRB1943, Anti-Notch1, clone 1A32 ZooMAb Rabbit Monoclonal detected Notch 1 protein from wild-type HEK293 cells (Lane 2), but not from Notch1 knockout cells (Lane 1). Each lane was loaded with 10 mg of cell lysate and Immobilon Signal Enhancer was used.
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of Notch1 peptide (Run 1) or non-specific control peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1943, Anti-Notch1, clone 1A32 ZooMAb® Rabbit Monoclonal. The binding of this antibody to Notch1 peptide displayed a KD value of 1.0 x 10-12 whereas its binding to non-specific control peptide displayed a KD of 1.6 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 | 1A32, 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 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 272.51 kDa |
| observed mol wt ~125 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_060087.3?  |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| species reactivity (predicted by homology) | mouse, rat |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunocytochemistry: suitable | |
| western blot: suitable | |
| UniProt accession no. | P46531 |
| Application: | Quality Control Testing Evaluated by Western Blotting in HEK293 cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Notch1 in HEK293 cell lysate. Tested applications Western Blotting Analysis: A 1:1000 dilution from a representative lot detected Notch1 from wild-type HEK293 cells, but not from Notch1 knockout cells. Immunocytochemistry Analysis: A 1:200 dilution from a representative lot detected Notch1 in HeLa and A431 cells. Affinity Binding Assay: A representative lot of this antibody bound Notch1 peptide with a KD of 1.0 x 10-12 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 |
| Application: | Anti-Notch1, clone 1A32 ZooMAb®, Cat. No. ZRB1943, is a recombinant Rabbit monoclonal antibody that detects Notch1 and is tested for use in Affinity Binding Assay, Immunocytochemistry, 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. Learn more about ZooMAb here. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 13 amino acids from the extracellular domain within the C-terminal half of human Notch1. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 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: | 600 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 1A32 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects Notch1. It targets an epitope within 13 amino acids from the extracellular domain in the C-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: | Neurogenic locus notch homolog protein 1 (UniProt: P46531; also known as Notch 1, hN1, Translocation-associated notch protein TAN-1) is encoded by the NOTCH1 (also known as TAN1) gene (Gene ID: 4851) in human. Notch1 is a single-pass type I membrane protein that is synthesized with a signal peptide (aa 1-18), which is subsequently cleaved off to generate the mature form that contains a long extracellular domain (aa 19-1735), a transmembrane domain (aa 1736-1756), and a cytoplasmic domain (aa 1757-2555). It is expressed in most fetal tissues and is found in abundance in spleen, brain stem, and lung tissue. It is also found in most adult lymphoid tissues. Notch1 is synthesized in the endoplasmic reticulum in an inactive form and 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. Cleavage results in a C-terminal fragment N(TM) and a N-terminal fragment N(EC). Following ligand binding, it is cleaved by ADAM17 to yield a membrane-associated intermediate fragment called notch extracellular truncation. Following endocytosis, this fragment is then cleaved by one of the catalytic subunits of g-secretase to release a Notch-derived peptide containing the intracellular domain (NICD) from the membrane. Notch 1 functions as a receptor for membrane-bound ligands Jagged-1 (JAG1), Jagged-2 (JAG2) and Delta-1 (DLL1) to regulate cell-fate determination. Upon ligand activation through the released NICD, it forms a transcriptional activator complex with RBPJ/RBPSUH and activates genes of the enhancer of split locus. Notch 1 is reported to be involved in angiogenesis and negatively regulates endothelial cell proliferation and migration and angiogenic sprouting. It is also involved in the maturation of both CD4+ and CD8+ cells in the thymus. Notch1 represses neuronal and myogenic differentiation and plays a role in post-implantation development, including mesoderm development, somite formation and neurogenesis. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| Storage Temp. | 2-8°C |