Human CBR1 knockout A549 cell line
- Advanced Validation
- What is this?
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CBR1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
15-hydroxyprostaglandin dehydrogenase [NADP+], CBR1_HUMAN, CRN, Carbonyl Reductase 1, Carbonyl reductase [NADPH] 1, NADPH-dependent carbonyl reductase 1, Prostaglandin 9-ketoreductase, Prostaglandin-E(2) 9-reductase, SDR21C1
- WB
Lab
Western blot - Human CBR1 knockout A549 cell line (AB266954)
Lanes 1-3 : Merged signal (red and green). Green - ab156590 observed at 30 kDa. Red - loading control ab8245 observed at 36 kDa.
ab156590 Anti-CBR1 antibody [EPR9660] was shown to specifically react with CBR1 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266954 (knockout cell lysate ab257874) was used. Wild-type and CBR1 knockout samples were subjected to SDS-PAGE. ab156590 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-CBR1 antibody [EPR9660] (<a href='/en-us/products/primary-antibodies/cbr1-antibody-epr9660-ab156590'>ab156590</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
CBR1 knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human CBR1 knockout A549 cell line (ab266954)
Lane 3:
HeLa cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 30 kDa
Observed band size: 30 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CBR1 knockout A549 cell line (AB266954)
Homozygous : 1 bp insertion in exon1
Reactivity data
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.
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.
- 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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Carbonyl reductase 1 functions in the metabolism of prostaglandins steroids and aromatic aldehydes. Cbr1 is not known to form part of a larger protein complex but acts independently to execute its enzymatic roles. It exhibits broad substrate specificity allowing it to reduce a variety of carbonyl groups which influences several physiological processes such as lipid metabolism and hormone synthesis.
Pathways
The enzyme plays an important role in the prostaglandin metabolic pathway where it reduces prostaglandin E2. Additionally Cbr1 interacts within the retinol metabolism pathway. In these pathways it functions alongside proteins such as aldo-keto reductases and aldose reductase sharing substrate specificity which impacts cellular responses and metabolic regulation.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CBR1 antibody [EPR9660] (AB156590)](https://content.abcam.com/products/images/cbr1-antibody-epr9660-ab156590--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img129860.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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