Human POR (Cytochrome P450 Reductase) knockout HeLa cell line
- Advanced Validation
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POR KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4.
View Alternative Names
CPR, CYPOR, Cytochrome p450 oxidoreductase, DKFZp686G04235, FLJ26468, NADPH dependent cytochrome P450 reductase, NADPH--cytochrome P450 reductase, NCPR_HUMAN, P450 (cytochrome) oxidoreductase, P450 Cytochrome Oxidoreductase, P450R, POR
- WB
Lab
Western blot - Human POR (Cytochrome P450 Reductase) knockout HeLa cell line (AB264996)
ab180597 Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] was shown to specifically react with Cytochrome P450 Reductase in HeLa wild-type cells ( ab255448). 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° at 1 in 10000 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.
All lanes:
Western blot - Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] (<a href='/en-us/products/primary-antibodies/cytochrome-p450-reductase-antibody-epr14479b-ab180597'>ab180597</a>) at 1/10000 dilution
Lane 1:
Wild-type HeLa lysate at 20 µg
Lane 2:
Cytochrome P450 Reductase knockout HeLa lysate at 20 µg
Lane 2:
Western blot - Human POR (Cytochrome P450 Reductase) knockout HeLa cell line (ab264996)
Lane 3:
HepG2 lysate at 20 µg
Lane 4:
A431 lysate at 20 µg
Predicted band size: 76 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human POR (Cytochrome P450 Reductase) knockout HeLa cell line (AB264996)
Homozygous : Insertion of the selection cassette in exon 4.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
CPR facilitates electron donation to all microsomal cytochrome P450 enzymes which are involved in a variety of oxidative reactions. This protein is essential for P450 activity assay as it is part of a multienzyme complex that metabolizes exogenous and endogenous compounds efficiently. Without cytochrome P450 reductase the catalytic activity of cytochrome P450 enzymes becomes impaired affecting metabolizing mechanisms for various substances and having systemic biological impacts.
Pathways
Cytochrome P450 reductase directly participates in drug metabolism and steroid biosynthesis pathways. It together with the cytochrome P450 enzymes metabolizes drugs to make them more water-soluble for excretion. The proteins 3 beta-hydroxysteroid dehydrogenase and CYP3A4 are some of the related components within these pathways working in conjunction to achieve metabolic homeostasis. P450 reductase assays are often utilized to study these pathways further understanding the biological roles and interactions of these proteins.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB180597
Anti-Cytochrome P450 Reductase antibody [EPR14479(B)]
BOND RX™ Validated|RabMAb|Recombinant|KO Validated|20ul selling size
![Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] (AB180597)](https://content.abcam.com/products/images/cytochrome-p450-reductase-antibody-epr14479b-ab180597--immunocytochemistry-immunofluorescence-img120459.jpg)
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Product promise
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