Human APP (Amyloid Precursor Protein) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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APP KO cell line available to order. KO validated by Immunocytochemistry, 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 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
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
- ICC/IF
Unknown
Immunocytochemistry/ Immunofluorescence - Human APP (Amyloid Precursor Protein) knockout HEK-293T cell line (AB255362)
ab32136 staining Amyloid Precursor Protein in wild-type HEK293 cells (top panel) and APP knockout HEK293 cells (ab255362) (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32136 at 1/500 dilution and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
- WB
Unknown
Western blot - Human APP (Amyloid Precursor Protein) knockout HEK-293T cell line (AB255362)
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 (ab255362)
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
false
- Sanger seq
Unknown
Sanger Sequencing - Human APP (Amyloid Precursor Protein) knockout HEK-293T cell line (AB255362)
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
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.
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'.
For licensing inquiries, please contact partnerships@abcam.com