Product nameHuman POR (Cytochrome P450 Reductase) knockout HeLa cell line
Parental Cell LineHeLa
Mutation descriptionKnockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4
Knockout validationSanger Sequencing, Western Blot (WB)
Tested applicationsSuitable for: WBmore details
Recommended control: Human Wild Type HeLa cell line (ab255448)
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
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
FunctionThis enzyme is required for electron transfer from NADP to cytochrome P450 in microsomes. It can also provide electron transfer to heme oxygenase and cytochrome B5.
Involvement in diseaseDefects in POR are the cause of adrenal hyperplasia variant type (AHV) [MIM:201750]; also known as Antley-Bixler syndrome-like phenotype with disordered steroidogenesis. AHV is a rare variant of congenital adrenal hyperplasia. It is an autosomal recessive disorder with apparent combined P450C17 and P450C21 deficiency. Affected girls are born with ambiguous genitalia, but their circulating androgens are low and virilization does not progress. Conversely, affected boys are sometimes born undermasculinized. Boys and girls can also present with bone malformations, in some cases resembling the pattern seen in patients with Antley-Bixler syndrome.
Defects in POR are a cause of isolated disordered steroidogenesis (IDS) [MIM:201750].
Sequence similaritiesIn the C-terminal section; belongs to the flavoprotein pyridine nucleotide cytochrome reductase family.
Contains 1 FAD-binding FR-type domain.
Contains 1 flavodoxin-like domain.
Cellular localizationEndoplasmic reticulum membrane. Anchored to the ER membrane by its N-terminal hydrophobic region.
- Information by UniProt
KO cell lysates
Our Abpromise guarantee covers the use of ab264996 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 76 kDa.|
All lanes : Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] (ab180597) at 1/10000 dilution
Lane 1 : Wild-type HeLa lysate
Lane 2 : Cytochrome P450 Reductase knockout HeLa lysate
Lane 3 : HepG2 lysate
Lane 4 : A431 lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 76 kDa
ab180597 Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] was shown to specifically react with Cytochrome P450 Reductase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264996 (knockout cell lysate ab257595) was used. Wild-type and Cytochrome P450 Reductase knockout samples were subjected to SDS-PAGE. ab180597 and Anti-GAPDH antibody [6C5] - Loading Control?(ab8245) were incubated overnight at 4^°C at 1 in 10000 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 4.
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.
ab264996 has not yet been referenced specifically in any publications.