NOTCH1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 5.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 5.
Alternative names
9930111A19Rik, AOS5, AOVD1, Lin-12, Motch A, NICD, NOTC1_HUMAN, NOTCH, NOTCH, Drosophila, homolog of, 1, Neurogenic locus notch homolog protein 1, Neurogenic locus notch protein homolog, Notch 1, Notch 1 intracellular domain, Notch gene homolog 1 (Drosophila), Notch homolog 1, translocation-associated (Drosophila), TAN1, Translocation associated notch homolog, Translocation-associated notch protein TAN-1, mIS6, mT14, xotch
What's included?
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NOTCH1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 5.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 5.
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- NOTCH1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
Storage
- Shipped at conditions
- Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Notch1 also known as Notch-1 is a transmembrane receptor involved in cell fate decisions. It belongs to the Notch protein family and has a molecular weight of around 270 kDa. Notch1 is expressed widely particularly in tissues such as blood cells and neuronal tissue. The receptor consists of extracellular EGF-like repeats a transmembrane domain and an intracellular domain that translocates to the nucleus upon activation. Notch1 plays a critical role in intercellular communication through ligand engagement which then triggers a series of proteolytic cleavages that release the Notch intracellular domain (NICD) for nuclear translocation.
- Biological function summary
Notch1 is essential in various cellular processes including differentiation proliferation and apoptosis. It functions as part of a complex signaling pathway that regulates these processes. Notch1 interacts with other elements like Jagged and Delta/Serrate/LAG-2 (DSL) family ligands to control gene expression patterns that determine cell lineage outcomes. This interaction affects the development of many systems such as the immune and nervous systems. Consequently Notch1 significantly influences the formation of organs and the maintenance of stem cell populations.
- Pathways
Notch1 plays a significant role in both the Notch signaling and the Wnt signaling pathways. In the Notch signaling pathway Notch1 upon ligand binding partners with CSL (CBF1/RBP-Jκ in mammals) and Mastermind-like proteins for transcriptional regulation. This pathway interlinks with the Wnt pathway that involves proteins like β-catenin affecting the regulation of gene transcription. The interplay between Notch1 and these pathways is fundamental in determining outcomes in cell proliferation and differentiation emphasizing the interconnected nature of signaling networks.
- Associated diseases and disorders
Notch1 is associated with T-cell acute lymphoblastic leukemia and breast cancer. Altered Notch1 signaling often through gain-of-function mutations can drive oncogenesis by disrupting normal cell differentiation and promoting uncontrolled proliferation. In T-cell acute lymphoblastic leukemia aberrant activation of Notch1 leads to increased expression of target genes working closely with related proteins like c-Myc. In breast cancer dysregulation of Notch1 signaling may facilitate tumor growth and metastasis indicating its role as a therapeutic target.
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3 product images
Western blot - Human NOTCH1 knockout HeLa cell lysate (ab257006)
Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: NOTCH1 knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - Anti-Notch1 antibody [EP1238Y] ab52627 observed at 110-120 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-Notch1 antibody [EP1238Y] ab52627 Anti-Notch1 antibody [EP1238Y] was shown to specifically react with Notch1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human NOTCH1 knockout HeLa cell line ab261762 (knockout cell lysate ab257006) was used. Wild-type and Notch1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-Notch1 antibody [EP1238Y] ab52627 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.All lanes: Western blot - Anti-Notch1 antibody [EP1238Y] (Anti-Notch1 antibody [EP1238Y] ab52627) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: Western blot - Human NOTCH1 knockout HeLa cell line (Human NOTCH1 knockout HeLa cell line ab261762)
Lane 2: NOTCH1 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 272 kDa
Observed band size: 110-120 kDa
Western blot - Human NOTCH1 knockout HeLa cell lysate (ab257006)
Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: NOTCH1 knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab65297 observed at 110 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
ab65297 Anti-Notch1 antibody was shown to specifically react with Notch1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human NOTCH1 knockout HeLa cell line ab261762 (knockout cell lysate ab257006) was used. Wild-type and Notch1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab65297 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.All lanes: Anti-Notch1 antibody (ab65297) at 1 µg/mL
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: NOTCH1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human NOTCH1 knockout HeLa cell line (Human NOTCH1 knockout HeLa cell line ab261762)
Performed under reducing conditions.
Sanger Sequencing - Human NOTCH1 knockout HeLa cell lysate (ab257006)
Homozygous: 1 bp insertion in exon 5
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