| 货号 | AF3647-SP |
| 别名 | EC 2.1.2.11; EC 3.4.21.68; hN1; Notch (Drosophila) homolog 1 (translocation-associated); notch 1Notch homolog 1, translocation-associated (Drosophila); Notch homolog 1, translocation-associated; TAN1; TAN1neurogenic locus notch homolog protein 1; Translocation-associated notch protein TAN-1 |
| 反应种属 | Human |
| 应用 | Western Blot(1 µg/mL) Flow Cytometry(2.5 µg/106cells) Chromatin Immunoprecipitation (ChIP)(5 µg/5 x 106cells) Immunocytochemistry(5-15 µg/mL) |
| 目标/特异性 | Detects the intracellular domain (ICD) of human Notch-1 in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 1% cross-reactivity with recombinant human (rh) Notch-2 ICD, rhNotch-3 ICD, and rhNotch-4 ICD is observed. |
| 使用方法 | Western Blot: 1 µg/mL Flow Cytometry: 2.5 µg/106cells Chromatin Immunoprecipitation (ChIP): 5 µg/5 x 106cells Immunocytochemistry: 5-15 µg/mL |
| 来源 | Reconstitute at 0.2 mg/mL in sterile PBS. |
| 产品组分 |
| 供应商 | R&D Systems |
| Entrez Gene IDs | 4851 (Human); 25496 (Rat) |
| 应用文献 | |
| R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions. A dual molecular analogue tuner for dissecting protein function in mammalian cells | |
| 纯化方式 | Antigen Affinity-purified |
| 免疫原 | E. coli-derived recombinant human Notch‑1 Gly2428-Lys2556 Accession # P46531 |
| 生物活性 | Human |
| 标记 | Unconjugated |
| 溶解方法 | Reconstitute at 0.2 mg/mL in sterile PBS. |
| 背景 | Notch-1 (so named for "notches" in fly wings; also TAN-1) is a 300 kDa member of the Notch family of glycoproteins. It is associated with gene activation in both embryo and adult. Human Notch-1 is a 2538 amino acid (aa) type I transmembrane glycoprotein. It undergoes Golgi processing to generate a heterodimer composed of a 180‑200 kDa disulfide-linked extracellular domain (aa 18‑1664) and a 120 kDa membrane-bound segment (aa 1665‑2556). Upon ligand binding, the 110 kDa segment undergoes two cleavages which generate an NICD (notch intracellular domain) (aa 1754‑2556), a nuclear transcription factor. One isoform shows a deletion of aa 248‑288. Over aa 2428‑2556, human Notch 1 is 83% and 89% aa identical to canine and mouse Notch-1, respectively. |
| 运输条件 | Blue Ice |
| 存放说明 | 4℃ |
| 参考文献 |
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Detection of Human Notch-1. Western blot shows lysates of MOLT‑4 human acute lymphoblastic leukemia cell line, Jurkat human acute T cell leukemia cell line, RPMI 8226 human multiple myeloma cell line, 293T human embryonic kidney cell line, and 293T human embryonic kidney cell line (1 µg per lane), transfected with full length human Notch‑1. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Notch‑1 Intracellular Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3647) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for Notch-1 intracellular domain (NICD) and full length Notch‑1 (Notch‑1 FL) at approximately 115 and 300 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1. | |
Detection of Notch‑1-regulated Genes by Chromatin Immunoprecipitation. Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin for 30 minutes was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Notch‑1/DNA complexes were immunoprecipitated using 5 μg Sheep Anti-Human Notch‑1 Intracellular Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3647) or control antibody (Catalog # 5-001-A) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Sheep IgG Secondary Antibody (Catalog # BAF016). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The c-myc promoter was detected by standard PCR. | |
Notch‑1 in Saos-2 Human Cell Line. Notch‑1 was detected in immersion fixed Saos-2 human osteosarcoma cell line using 10 µg/mL Sheep Anti-Human Notch‑1 Intracellular Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3647) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips. |
