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NCSTN KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp deletion after Asn164 of the WT protein
Frameshift = 100%.

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Images

Next Generation Sequencing - Human NCSTN (Nicastrin) knockout HeLa cell line (AB273849), expandable thumbnail

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

Knockout validation

Next Generation Sequencing

Mutation description

Knockout achieved by CRISPR/Cas9 X = 1 bp deletion after Asn164 of the WT protein
Frameshift = 100%

Alternative names

Recommended products

NCSTN KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp deletion after Asn164 of the WT protein
Frameshift = 100%.

Key facts

Cell type

HeLa

Form

Liquid

Mutation description

Knockout achieved by CRISPR/Cas9 X = 1 bp deletion after Asn164 of the WT protein
Frameshift = 100%

Disease

Adenocarcinoma

Concentration
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Properties

Gene name

NCSTN

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Next Generation Sequencing

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

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.

Storage

Shipped at conditions

Dry Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

Notes

Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab271142). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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

Nicastrin also known as NAS is an integral component of the gamma-secretase complex. This protein interacts mechanically by assisting in substrate recognition and stabilization within the complex. Nicastrin has a molecular weight of approximately 75 kDa. It is predominantly expressed in various tissues including the brain where it plays significant role in neuronal functions.

Biological function summary

Nicastrin functions as a part of the gamma-secretase complex which also includes presenilin anterior pharynx defective 1 (APH-1) and presenilin enhancer 2 (PEN-2). This complex is essential for intramembrane proteolysis of type I membrane proteins such as amyloid precursor protein (APP) and Notch. The activity of nicastrin influences processes like synaptic organization and neuronal development.

Pathways

Gamma-secretase activity facilitated by nicastrin is central in the Notch signaling pathway and the amyloidogenic pathway. In the Notch pathway it helps release the Notch intracellular domain important for T-cell development. In the amyloidogenic pathway the complex processes APP contributing to amyloid-beta peptide formation. Beta-secretase (BACE1) also plays an important role in this pathway acting upstream of gamma-secretase.

Associated diseases and disorders

The imbalance in nicastrin function connects it to Alzheimer's disease and certain cancers. Its role in amyloid-beta peptide production links nicastrin to Alzheimer's disease pathogenesis. In cancer the deregulation of Notch signaling in which nicastrin is involved influences tumor progression and survival. The interaction of nicastrin with presenilin is particularly notable in these conditions further emphasizing its significance in disease mechanisms.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

1 product image

  • Next Generation Sequencing - Human NCSTN (Nicastrin) knockout HeLa cell line (ab273849), expandable thumbnail

    Next Generation Sequencing - Human NCSTN (Nicastrin) knockout HeLa cell line (ab273849)

    1 bp deletion after Asn164 of the WT protein

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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