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AB269476

Human CYP1B1 knockout A549 cell line

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CYP1B1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 2 bp deletion; Frameshift: 100%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
2 Images
Western blot - Human CYP1B1 knockout A549 cell line (AB269476)
  • WB

Lab

Western blot - Human CYP1B1 knockout A549 cell line (AB269476)

Western blot : Anti-CYP1B1 antibody [EPR14972] (ab185954) staining at 1/5000 dilution, shown in black; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab185954 was shown to bind specifically to CYP1B1. A band was observed at 55 kDa in wild-type A549 cell lysates with no signal observed at this size in CYP1B1 knockout cell line ab269476 (knockout cell lysate ab269641). To generate this image, wild-type and CYP1B1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with a high-sensitivity ECL substrate kit and imaged with 8 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-CYP1B1 antibody [EPR14972] - C-terminal (<a href='/en-us/products/primary-antibodies/cyp1b1-antibody-epr14972-c-terminal-ab185954'>ab185954</a>) at 1/5000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

CYP1B1 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human CYP1B1 knockout A549 cell lysate (<a href='/en-us/products/cell-lysates/human-cyp1b1-knockout-a549-cell-lysate-ab269641'>ab269641</a>)

Lane 2:

Western blot - Human CYP1B1 knockout A549 cell line (ab269476)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Lane 5:

SiHa cell lysate at 20 µg

Lane 6:

Human Brain cell lysate at 20 µg

Lane 7:

Human Kidney cell lysate at 20 µg

Lane 8:

Human Liver cell lysate at 20 µg

Lane 9:

HepG2 cell lysate at 20 µg

Lane 10:

LoVo cell lysate at 20 µg

Secondary

All lanes:

HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 61 kDa

Observed band size: 55 kDa

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Next Generation Sequencing - Human CYP1B1 knockout A549 cell line (AB269476)
  • NGS

Supplier Data

Next Generation Sequencing - Human CYP1B1 knockout A549 cell line (AB269476)

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 2 bp deletion; Frameshift : 100%

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion, 2 bp deletion; Frameshift: 100%

Disease

Carcinoma

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Complementary Products

Primary Antibodies

AB185954

Anti-CYP1B1 antibody [EPR14972] - C-terminal

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP1B1 antibody [EPR14972] - C-terminal (AB185954)

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Cell Lines & Lysates

AB269641

Human CYP1B1 knockout A549 cell lysate

Western blot - Human CYP1B1 knockout A549 cell lysate (AB269641)

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Primary Antibodies

AB22717

Anti-Cytochrome P450 1A2 antibody [d15 (16VII F10F12)]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome P450 1A2 antibody [d15 (16VII F10F12)] (AB22717)

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Primary Antibodies

AB126887

Anti-CYP1A1 antibody

Western blot - Anti-CYP1A1 antibody (AB126887)

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Product details

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.

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What's included?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab269476-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab269476 Human CYP1B1 knockout A549 cell line", "number":"AB269476-CMP01" }, { "size":"1 x 1000000 Cells/vial", "name":"ab259777 Human wild-type A549 cell line", "number":"AB269476-CMP02" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab269476-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab269476 Human CYP1B1 knockout A549 cell line", "number":"AB269476-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
CYP1B1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium

F-12K + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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Supplementary information

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

CYP1B1 also known as cytochrome P450 1B1 is a member of the cytochrome P450 superfamily of enzymes. This monoxygenase enzyme with a molecular mass of approximately 60 kDa plays a role in the metabolism of xenobiotics and endogenous compounds. It is mainly expressed in various tissues including the liver lungs and the artery walls. The enzyme catalyzes the hydroxylation of aromatic hydrocarbons aiding in their detoxification and removal from the body.
Biological function summary

CYP1B1 contributes to the metabolism of various drugs and toxins helping to maintain cellular homeostasis. This enzyme also participates in the conversion of steroids and fatty acids impacting hormonal regulation and fatty acid balance. While CYP1B1 does not form a specific complex it functions within the cytochrome P450 enzyme system which collectively metabolizes numerous compounds. This functional integration within a broad enzymatic network highlights its role in chemical transformation processes.

Pathways

CYP1B1 serves an essential role in the Phase I drug metabolism pathway. This pathway includes the functionalization of xenobiotic substances to prepare them for Phase II reactions. The enzyme is involved in the hydroxylation of substrates which are important steps in both the AhR and chemical carcinogenesis pathways. CYP1B1 interacts with other cytochrome P450 family members such as CYP1A1 which further facilitates the detoxification and clearance of harmful compounds from the body.

CYP1B1 is significantly associated with primary congenital glaucoma and various cancers such as breast cancer. In these conditions altered CYP1B1 activity can lead to atypical metabolism of endogenous hormones contributing to disease progression. CYP1B1's interaction with cyclin D1 is noted in cancer pathogenesis where dysregulation of cell cycle progression occurs. Therefore understanding CYP1B1's expression and function can provide insights into therapeutic strategies targeting these disorders.
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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

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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