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AB258061

Human NCSTN (Nicastrin) knockout HEK-293T cell lysate

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NCSTN 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 1.

View Alternative Names

APH 2, ATAG1874, Anterior pharynx defective 2, KIAA0253, NCT, NICA_HUMAN, Ncstn, Nicastrin, RP11 517F10.1, RP11517F101

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Western blot - Human NCSTN (Nicastrin) knockout HEK-293T cell lysate (AB258061)
  • WB

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Western blot - Human NCSTN (Nicastrin) knockout HEK-293T cell lysate (AB258061)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : NCSTN knockout HEK293T cell lysate (20 ug)
Lane 3 : HeLa cell lysate (20 ug)
Lane 4 : HL-60 cell lysate (20 ug)

ab68145 was shown to specifically react with Nicastrin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266602 (knockout cell lysate ab258061) was used. Wild-type and Nicastrin knockout samples were subjected to SDS-PAGE. ab68145 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC at 1 in 1000 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.

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Sanger Sequencing - Human NCSTN (Nicastrin) knockout HEK-293T cell lysate (AB258061)
  • Sanger seq

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Sanger Sequencing - Human NCSTN (Nicastrin) knockout HEK-293T cell lysate (AB258061)

Homozygous : 1 bp insertion in exon 1

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
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.

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What's included?

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Properties and storage information

Gene name
NCSTN
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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