Human NOTCH1 knockout HeLa cell line
- Advanced Validation
- What is this?
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NOTCH1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 5. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View 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
- WB
Lab
Western blot - Human NOTCH1 knockout HeLa cell line (AB261762)
Lanes 1- 2 : Merged signal (red and green). Green - ab52627 observed at 110-120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab52627 was shown to react with Notch1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261762 (knockout cell lysate ab257006) was used. Wild-type HeLa and NOTCH1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab52627 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and 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] (<a href='/en-us/products/primary-antibodies/notch1-antibody-ep1238y-ab52627'>ab52627</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human NOTCH1 knockout HeLa cell line (ab261762)
Lane 2:
NOTCH1 knockout HeLa cell lysate at 20 µg
Predicted band size: 272 kDa
Observed band size: 110-120 kDa
false
- ChIC/CUT&RUN-seq
Supplier Data
ChIC/CUT&RUN sequencing - Human NOTCH1 knockout HeLa cell line (AB261762)
CUT&RUN profiling with Notch1 antibody reveals the expected genomic enrichment pattern in wild-type (WT) cells, which is substantially reduced in NOTCH1 knockout (KO) cells. Representative genome browser tracks show CUT&RUN data generated using the CUTANA™ CUT&RUN Kit (EpiCypher 14-1048) with Notch1 antibody (Abcam ab52627, 0.5 µg). 500,000 HeLa WT or KO (ab261762) cells were used per reaction. IgG, H3K4me3, and H3K27me3 antibodies were included as controls to assess non-specific background, active promoters, and repressed chromatin, respectively. Libraries were prepared using the CUTANA™ CUT&RUN Library Prep Kit (EpiCypher 14-1001). Sequencing was performed with paired-end 50 bp reads, and data were processed on CUTANA™ Cloud (cloud.epicypher.com) by alignment to the hg38 genome. Images were generated using Integrative Genomics Viewer (IGV, Broad Institute).
- ChIC/CUT&RUN-seq
Supplier Data
ChIC/CUT&RUN sequencing - Human NOTCH1 knockout HeLa cell line (AB261762)
CUT&RUN profiling with Notch1 antibody demonstrates robust genome-wide enrichment in wild-type (WT) cells, which is markedly diminished in NOTCH1 knockout (KO) cells. Heatmaps of genome-wide signal flanking annotated transcription start sites (TSSs, +/- 2 kbp) display CUT&RUN data generated using the CUTANA™ CUT&RUN Kit (EpiCypher 14-1048) with Notch1 antibody (Abcam ab52627, 0.5 µg). 500,000 HeLa WT or KO (Abcam ab261762) cells were used per reaction. IgG antibody was included as a negative control to assess non-specific background. Libraries were prepared using the CUTANA™ CUT&RUN Library Prep Kit (EpiCypher 14-1001). Sequencing was performed with paired-end 50 bp reads, and data were processed on CUTANA™ Cloud (cloud.epicypher.com) by alignment to the hg38 genome. Heatmaps were generated using deepTools (Ramнrez et al., Nucleic Acids Res. 2014; PMID : 24799436). Row-linked data are ranked by intensity relative to Notch1 WT, with red indicating high localized enrichment and blue denoting background. Validated antibodies show genome-wide enrichment above IgG background consistent with Notch1 binding in WT cells and near complete loss of signal in KO cells.
- WB
Lab
Western blot - Human NOTCH1 knockout HeLa cell line (AB261762)
Lanes 1- 2 : Merged signal (red and green). Green - ab65297 observed at 110 kDa. Red - 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 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 (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 (ab216773) and 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 (<a href='/en-us/products/unavailable/notch1-antibody-ab65297'>ab65297</a>) 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 (ab261762)
false
- Sanger seq
Unknown
Sanger Sequencing - Human NOTCH1 knockout HeLa cell line (AB261762)
Homozygous : 1 bp insertion in exon 5.
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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