Human SCD (SCD1) knockout HeLa cell line (ab265220)
Properties
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Number of cells
1 x 106 cells/vial, 1 mL -
Viability
~90% -
Adherent /Suspension
Adherent -
Tissue
Cervix -
Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10 -
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 SCD (SCD1) knockout HeLa cell line (ab265220)All lanes : Anti-SCD1 antibody [CD.E10] (ab19862) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SCD knockout HeLa cell lysate
Lane 3 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 36 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab19862 observed at 36 kDa. Red - loading control ab2866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
ab19862 was shown to react with SCD1 in HeLa wild-type cells in western blot with loss of signal observed in SCD knockout cell line ab265220 (SCD knockout cell lysate ab257658). HeLa wild-type and SCD knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab19862 and ab2866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Human SCD knockout HeLa cell line (ab265220)All lanes : Anti-SCD1 antibody [EPR21963] (ab236868) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SCD knockout HeLa cell lysate
Lane 3 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 42 kDaanes 1-3: Merged signal (red and green). Green - ab236868. Red - loading control ab8245 observed at 50 kDa.
ab236868 Anti-SCD1 antibody [EPR21963] was shown to specifically react with SCD1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265220 (knockout cell lysate ab257658) was used. Wild-type and SCD1 knockout samples were subjected to SDS-PAGE. ab236868 and Anti-tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C 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 10000 dilution for 1 hour at room temperature before imaging.
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Sanger Sequencing - Human SCD knockout HeLa cell line (ab265220)Allele-1: 4 bp deletion in exon 3.
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Sanger Sequencing - Human SCD knockout HeLa cell line (ab265220)Allele-2: 4 bp deletion in exon 3.
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Sanger Sequencing - Human SCD knockout HeLa cell line (ab265220)Allele-3: 4 bp deletion in exon 3.
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Sanger Sequencing - Human SCD knockout HeLa cell line (ab265220)Allele-5: Insertion of the selection cassette in exon 3.
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Sanger Sequencing - Human SCD knockout HeLa cell line (ab265220)Allele-4: Insertion of the selection cassette in exon 3.
