Human CCND1 (Cyclin D1) knockout HeLa cell line (ab255348)
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 -
Purity
Immunogen affinity purified -
Research areas
Images
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Western blot - Human CCND1 (Cyclin D1) knockout HeLa cell line (ab255348)All lanes : Anti-Cyclin D1 antibody [EP272Y] (ab40754) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CCND1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 36 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab40754 observed at 36 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.
ab40754 was shown to react with CCND1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255348 (knockout cell lysate ab263808) was used. Wild-type HeLa and CCND1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab40754 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 1000 dilution and a 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 CCND1 (Cyclin D1) knockout HeLa cell line (ab255348)All lanes : Anti-Cyclin D1 antibody [SP4] (ab16663) at 1/200 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CCND1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 36 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab16663 observed at 36 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.
ab16663 was shown to react with CCND1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255348 (knockout cell lysate ab263808) was used. Wild-type HeLa and CCND1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab16663 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 200 dilution and a 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 CCND1 knockout HeLa cell line (ab255348)Homozygous: 5 bp deletion in exon 1.