Human APP (Amyloid Precursor Protein) knockout HEK-293T cell line (ab255362)
Properties
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Number of cells
1 x 106 cells/vial, 1 mL -
Viability
~90% -
Adherent /Suspension
Adherent -
Tissue
Kidney -
Cell type
epithelial -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12 -
Mycoplasma free
Yes -
Storage instructions
Shipped on Dry Ice. Store in liquid nitrogen. -
Storage buffer
Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether -
Research areas
Images
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Western blot - Human APP knockout HEK293T cell line (ab255362)Lanes 1-2 : Anti-Amyloid Precursor Protein antibody [DE2B4] (ab12266) at 1/500 dilution
Lanes 3-4 : Anti-Amyloid Precursor Protein antibody [DE2B4] (ab12266)
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SH-SY5Y cell lysate
Lane 3 : Wild-type HEK-293T cell lysate
Lane 4 : APP knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 86 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Green signal. Green - ab12266 observed at 110 kDa.
Lanes 5 - 8: Red signal. Red - loading control, ab181602 observed at 37 kDa.
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.
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Flow Cytometry - Human APP (Amyloid Precursor Protein) knockout HEK293T 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). -
Sanger Sequencing - Human APP knockout HEK293T cell line (ab255362)Homozygous: 1 bp insertion in exon 5