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AB257596

Human POR (Cytochrome P450 Reductase) knockout HCT116 cell lysate

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POR KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon4.
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Western blot - Human POR (Cytochrome P450 Reductase) knockout HCT116 cell lysate (AB257596)
  • WB

Lab

Western blot - Human POR (Cytochrome P450 Reductase) knockout HCT116 cell lysate (AB257596)

Lane 1 : Wild-type HCT116 cell lysate (40µg)

Lane 2 : POR knockout HCT116 cell lysate (40µg)

Lane 3 : HepG2 cell lysate (40µg)

Lane 4 : A431 cell lysate (40µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab180597 observed at 80 kDa. Red - loading control ab8245 observed at 37 kDa.

ab180597 Rabbit monoclonal [EPR14479(B)] to Cytochrome P450 Reductase was shown to specifically react with Cytochrome P450 Reductase in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266889 (knockout cell lysate ab257596) was used. Wild-type and Cytochrome P450 Reductase knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab180597 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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 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 1/1000 dilution

Lane 1:

Wild-type HCT116 cell lysate at 40 µg

Lane 2:

POR knockout HCT116 cell lysate at 40 µg

Lane 2:

Western blot - Human POR (Cytochrome P450 Reductase) knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-por-cytochrome-p450-reductase-knockout-hct116-cell-line-ab266889'>ab266889</a>)

Lane 3:

HepG2 cell lysate at 40 µg

Lane 4:

A431 cell lysate at 40 µg

Predicted band size: 76 kDa

Observed band size: 80 kDa

false

Sanger Sequencing - Human POR (Cytochrome P450 Reductase) knockout HCT116 cell lysate (AB257596)
  • Sanger seq

Unknown

Sanger Sequencing - Human POR (Cytochrome P450 Reductase) knockout HCT116 cell lysate (AB257596)

Homozygous : Insertion of the selection cassette in exon4

Key facts

Cell type

HCT116

Species or organism

Human

Tissue

Colon

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon4.

Disease

Carcinoma

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
POR
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Cytochrome P450 reductase also known as NADPH--cytochrome P450 reductase (CPR) is an enzyme important in electron transfer. It has a mass of approximately 77 kDa and is responsible for transferring electrons from NADPH to cytochrome P450 enzymes supporting their function. The enzyme is predominantly expressed in the endoplasmic reticulum of liver cells and other tissues where detoxification processes occur. Cytochrome reductase as it is sometimes called plays a fundamental role in the metabolism of drugs and the synthesis of cholesterol steroids and other lipids.
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.

CPR has a significant connection to conditions like congenital adrenal hyperplasia and prostate cancer. Mutations or dysregulations in cytochrome P450 reductase can lead to accumulated steroid precursors and disrupted steroid hormone synthesis contributing to these disorders. The relationship with adrenal enzyme 21-hydroxylase and the androgen receptor illustrates the pathway complexities and the clinical relevance of cytochrome P450 reductase. Understanding these connections highlights the importance of CPR in maintaining normal physiological functions and how its abnormalities can lead to serious health issues.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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