NR2F2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
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
NR2F2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
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.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
Recommended control: Human wild-type HCT116 cell line (ab255451). 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.
NR2F2 also known as COUP-TFII (Chicken Ovalbumin Upstream Promoter Transcription Factor II) is a nuclear receptor that functions as a transcription factor. It has a molecular mass of approximately 48 kDa. It binds to DNA target sequences and regulates gene expression involved in various developmental processes. NR2F2 is expressed in the heart blood vessels and various tissues such as the lungs and the central nervous system. Its role in transcriptional regulation makes it important for proper cell and tissue differentiation.
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.
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.
NR2F2 is linked to congenital heart defects and cancer. Aberrant NR2F2 expression disrupts normal developmental processes contributing to heart malformations. In cancer altered NR2F2 expression and function can facilitate tumor progression and metastasis often involving interaction with other proteins like TGF-beta and SMADs essential for tumor suppression pathways. The ability of NR2F2 to control critical transcriptional networks makes it a significant target for understanding and potentially treating these conditions.
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Lanes 1- 2: Merged signal (red and green). Green - Anti-NR2F2 antibody [EPR18443] ab211777 observed at 45 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
Anti-NR2F2 antibody [EPR18443] 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 Human NR2F2 knockout HCT116 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. Anti-NR2F2 antibody [EPR18443] ab211777 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (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] (Anti-NR2F2 antibody [EPR18443] ab211777) 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)
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 45 kDa
Homozygous: 1 bp insertion in exon2
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