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AB263777

Human APP (Amyloid Precursor Protein) knockout HEK-293T cell lysate

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APP 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.

View Alternative Names

A BETA, A4 amyloid protein, A4_HUMAN, AAA, ABPP, AD1, AICD-50, AICD-57, AICD-59, AID(50), AID(57), AID(59), APP, APPI, Alzheimer disease amyloid protein, Amyloid beta (A4) precursor protein, Amyloid beta A4 protein, Amyloid beta protein, Amyloid intracellular domain 50, Amyloid intracellular domain 57, Amyloid intracellular domain 59, Amyloid precursor protein, Beta-APP40, Beta-APP42, Beta-amyloid precursor protein, C31, CTFgamma, CVAP, Cerebral vascular amyloid peptide, Gamma-CTF(50), Gamma-CTF(57), Gamma-CTF(59), PN 2, PN-II, PreA4, Protease nexin-II, S-APP-alpha, S-APP-beta, beta-amyloid peptide, peptidase nexin-II, sAPP

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Western blot - Human APP (Amyloid Precursor Protein) knockout HEK-293T cell lysate (AB263777)
  • WB

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Western blot - Human APP (Amyloid Precursor Protein) knockout HEK-293T cell lysate (AB263777)

Lanes 1 and 5 : HeLa cell lysate (20 μg)

Lanes 2 and 6 : SH-SY5Y cell lysate (20 μg)

Lanes 3 and 7 : Wild-type HEK-293T cell lysate (20 μg)

Lanes 4 and 8 : APP knockout HEK-293T cell lysate (20 μg)

ab12266 was shown to react with Amyloid Precursor Protein in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab255362 (knockout cell lysate ab263777) was used. Wild-type and Amyloid Precursor Protein knockout samples were subjected to SDS-PAGE. ab12266 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Anti-Amyloid Precursor Protein antibody [DE2B4] (<a href='/en-us/products/unavailable/amyloid-precursor-protein-antibody-de2b4-ab12266'>ab12266</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SH-SY5Y cell lysate at 20 µg

Lane 3:

Wild-type HEK-293T cell lysate at 20 µg

Lane 4:

APP knockout HEK-293T cell lysate at 20 µg

Lane 4:

Western blot - Human APP (Amyloid Precursor Protein) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-app-amyloid-precursor-protein-knockout-hek-293t-cell-line-ab255362'>ab255362</a>)

Secondary

All lanes:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/20000 dilution

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Sanger Sequencing - Human APP (Amyloid Precursor Protein) knockout HEK-293T cell lysate (AB263777)
  • Sanger seq

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Sanger Sequencing - Human APP (Amyloid Precursor Protein) knockout HEK-293T cell lysate (AB263777)

Homozygous : 1 bp insertion in exon 5

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 5.

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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
APP
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.

The Amyloid Precursor Protein (APP) also known as amyloid protein is a transmembrane protein that is approximately 695 to 770 amino acids in length depending on the isoform. The molecular mass of APP can vary but typically falls around 100 to 140 kDa. It is heavily expressed in the central nervous system particularly in neurons but also in other tissues like muscle and kidney. The APP undergoes proteolytic processing which leads to the generation of various fragments including beta-amyloid peptides.
Biological function summary

The processing of APP plays a fundamental role in neuronal growth survival and repair. APP is cleaved into fragments that can regulate synaptic function and plasticity. It does not operate as a part of a complex but interacts with various cellular components. The protein participates in signaling pathways influencing cellular adhesion motility and neurite outgrowth. APP’s numerous interaction partners facilitate its involvement in different cellular processes highlighting its critical role in normal cell function.

Pathways

The APP is a central component in the amyloidogenic pathway where its cleavage by beta-secretase and gamma-secretase yields beta-amyloid. This pathway is one of two primary metabolic routes for APP—alternative enzymatic processing through the non-amyloidogenic pathway precludes beta-amyloid formation releasing peptides that do not aggregate. Enzymes like BACE1 (beta-secretase 1) and presenilin are important in the amyloidogenic pathway directly resulting in the production of the neurotoxic amyloid beta-peptide.

APP is intensely linked to Alzheimer's disease and cerebral amyloid angiopathy. Accumulation of beta-amyloid peptides formed from APP cleavage is a hallmark of Alzheimer's disease leading to plaque formation in the brain. This aggregation impacts neuronal function and is associated with neurodegenerative processes. Interactions with proteins like tau are significant as tau also plays an essential role in Alzheimer's disease pathology. Misprocessing of APP and the resulting beta-amyloid aggregates are also contributors to cerebral amyloid angiopathy where deposits within cerebrovascular walls compromise vascular integrity.
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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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Complementary Products

Cell Lines & Lysates

AB255362

Human APP (Amyloid Precursor Protein) knockout HEK-293T cell line

Advanced Validation

Immunocytochemistry/ Immunofluorescence - Human APP (Amyloid Precursor Protein) knockout HEK-293T cell line (AB255362)

  • 0
  • 0
Primary Antibodies

AB32136

Anti-Amyloid Precursor Protein antibody [Y188]

BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Amyloid Precursor Protein antibody [Y188] (AB32136)

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Assay Kits

AB216944

Human Amyloid Precursor Protein ELISA Kit

Recombinant|SimpleStep

Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (AB216944)

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Proteins & Peptides

AB217407

Recombinant human Vitronectin/S-Protein (Animal Free)

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Product promise

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For full details, please see our Terms & Conditions

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

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