Human NR2F2 knockout HCT116 cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
NR2F2 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
Apolipoprotein A-I regulatory protein 1, COT2_HUMAN, COUP transcription factor 2, COUP transcription factor II, COUP-TF II, COUP-TF2, Nuclear receptor subfamily 2 group F member 2
- WB
Lab
Western blot - Human NR2F2 knockout HCT116 cell line (AB266888)
Lanes 1- 2 : Merged signal (red and green). Green - ab211777 observed at 45 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab211777 was shown to react with NR2F2 in wild-type HCT116 cells in western blot. The band observed in knockout cell line ab266888 (knockout cell lysate ab257186) lane below 45kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HCT116 and NR2F2 knockout HCT117 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab211777 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 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-NR2F2 antibody [EPR18443] (<a href='/en-us/products/primary-antibodies/nr2f2-antibody-epr18443-ab211777'>ab211777</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT116 cell lysate at 20 µg
Lane 2:
NR2F2 CRISPR/Cas9 edited HCT116 cell lysate at 20 µg
Lane 2:
Western blot - Human NR2F2 knockout HCT116 cell line (ab266888)
Predicted band size: 46 kDa
Observed band size: 45 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human NR2F2 knockout HCT116 cell line (AB266888)
Homozygous : 1 bp insertion in exon2
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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
McCoY5a + 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
NR2F2 is involved in the regulation of angiogenesis and cardiovascular development. It forms a heterodimer with retinoid X receptor (RXR) to regulate gene expression. This interaction places NR2F2 as part of a larger network controlling organ development and metabolic processes. NR2F2 activity also influences energy homeostasis and adipogenesis through its transcriptional regulation abilities.
Pathways
NR2F2 influences the Wnt signaling and Notch signaling pathways which play important roles in cell proliferation and differentiation. These pathways interact with other proteins such as beta-catenin and Notch receptors. NR2F2 modulates gene expression at different cellular stages and contributes to the precise regulation required within these pathways.
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
Primary Antibodies
AB211777
Anti-NR2F2 antibody [EPR18443]
RabMAb|Recombinant|Advanced Validation|20ul selling size|Trial Size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NR2F2 antibody [EPR18443] (AB211777)](https://content.abcam.com/products/images/nr2f2-antibody-epr18443-ab211777--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img110797.jpg)
- 5
- 2
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com