Product nameHuman PLAA knockout HeLa cell line
Parental Cell LineHeLa
Mutation descriptionKnockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 3
Knockout validationSanger Sequencing, Western Blot (WB)
Recommended control: Human Wild Type HeLa cell line (ab255928)
Please note a wild type cell line is not automatically included with a KO cell line order, if required please add recommended wild type cell line at no additional cost using the code WILDTYPE-TMTK1
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, supplemented with 10% (v/v) DMSO
Handling procedure: Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C will result in loss of viability.
1. Thaw the vial in 37°C water bath approximately 1-2 minutes.
2. Transfer the cell suspension to a 15 mL conical tube with pre-warmed 5 mL complete medium DMEM+10% FBS, spin 125×g for approximately 5 minutes at room temperature.
3. Resuspend the cell pellet with 1 mL pre-warmed complete medium DMEM+10% FBS and dispense into a 25 cm2 culture flask containing 10 mL pre-warmed complete complete medium DMEM+10% FBS.
4. Incubate the culture at 37°C incubator with 5% CO2.
5. A subcultivation ratio of 1:4-1:6 is recommended. Cells should be passaged when cells grow splitting at 80-90% confluence.
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.
Number of cells1 x 106 cells/vial, 1 mL
STR AnalysisAmelogenin 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
Antibiotic resistancePuromycin 1.00µg/ml
Storage instructionsShipped on Dry Ice. Store in liquid nitrogen.
Storage bufferConstituents: 8.7% DMSO, 2% Cellulose, methyl ether
FunctionInvolved in the maintenance of ubiquitin levels.
Sequence similaritiesBelongs to the WD repeat PLAP family.
Contains 6 ARM repeats.
Contains 1 PFU domain.
Contains 1 PUL domain.
Contains 7 WD repeats.
DomainThe PUL domain is composed of 6 armadillo-like repeats and mediates the interaction with VCP C-terminus.
The PFU domain mediates interaction with ubiquitin.
- Information by UniProt
KO cell lysates
All lanes : Anti-Phospholipase A2 activator protein (PLAP) antibody [EPR5481] (ab133589) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PLAA knockout HeLa cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : Daudi cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution
Observed band size: 87 kDa why is the actual band size different from the predicted?
ab133589 Anti-PLAA antibody [EPR5481] was shown to specifically react with PLAA in wild-type HeLa cells (ab255928). Loss of signal was observed when knockout cell line ab265255 (knockout cell lysate ab258121) was used. Wild-type and PLAA knockout samples were subjected to SDS-PAGE. ab133589 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4ºC 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 20000 dilution for 1 hour at room temperature before imaging.
Homozygous: Insertion of the selection cassette in exon 3.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab265255 has not yet been referenced specifically in any publications.