Human CDC42 knockout HEK-293T cell line (ab266522)
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 -
Antibiotic resistance
Puromycin 1.00µg/ml -
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 CDC42 knockout HEK293T cell line (ab266522)All lanes : Anti-CDC42 antibody [EPR15620] (ab187643) at 1/5000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : CDC42 knockout HEK-293T cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution
Predicted band size: 21 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab187643 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.
ab187643 Anti-CDC42 antibody [EPR15620] was shown to specifically react with CDC42 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266522 (knockout cell lysate ab256868) was used. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. ab187643 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 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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Western blot - Human CDC42 knockout HEK293T cell line (ab266522)All lanes : Anti-CDC42 antibody (ab64533) at 1 µg/ml
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : CDC42 knockout HEK-293T cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution
Predicted band size: 21 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab64533 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.
ab64533 Anti-CDC42 antibody was shown to specifically react with CDC42 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266522 (knockout cell lysate ab256868) was used. Wild-type and CDC42 knockout samples were subjected to SDS-PAGE. ab64533 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 µg/ml 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 CDC42 knockout HEK293T cell line (ab266522)Homozygous: 2 bp insertion in exon 3 -
Cell Culture - Human CDC42 knockout HEK293T cell line (ab266522)Representative images of CDC42 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
