ESRRA KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
- MCF7
- Species or organism
- Human
- Tissue
- Breast
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
Alternative names
ERR a, ERR-alpha, ERR1 protein, ERR1_HUMAN, ESRL 1, ESRR A, Estrogen receptor related 1, Estrogen receptor-like 1, Estrogen-related receptor alpha, Estrra, NR3B1, Nuclear receptor subfamily 3 group B member 1, Steroid hormone receptor ERR1, estrogen receptor related receptor alpha, hERR1
Recommended products
ESRRA KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
Key facts
- Cell type
- MCF7
- Species or organism
- Human
- Tissue
- Breast
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- ESRRA
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Female
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
- Slow to trypsinise.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 5-7x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Culture medium
- MEM + 10% FBS + 0.01 mg/ml bovine insulin
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
Recommended control: Human wild-type MCF7 cell line (Human wild-type MCF7 cell line ab288560). 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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Estrogen Related Receptor Alpha (ERRα) also known as NR3B1 is an orphan nuclear receptor with a molecular mass of approximately 51 kDa. It acts as a transcription factor and influences the expression of genes involved in energy metabolism. ERRα is expressed in numerous tissues such as the liver heart kidney and skeletal muscles. It binds to DNA as a monomer and alters specific gene expression activities. This protein lacks an identified ligand which classifies it as an orphan receptor but it shares structural similarities with estrogen receptors allowing the regulation of gene transcription in several physiological contexts.
- Biological function summary
Estrogen Related Receptor Alpha regulates cellular energy production processes including mitochondrial biogenesis and oxidative phosphorylation. It plays a role in the biological function of energy homeostasis and increases the expression of genes related to metabolic processes. ERRα operates as part of a transcriptional regulatory network and interacts with other nuclear receptors and coactivators to modulate its activity. It often forms complexes with coactivators such as peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) to exert its effects on mitochondrial function.
- Pathways
ERRα is integral to the PGC-1α signaling pathway and the AMPK (AMP-activated protein kinase) signaling pathway. These pathways are important in regulating energy metabolism and adaptive thermogenesis. ERRα interacts with PGC-1α to enhance mitochondrial function and energy expenditure while AMPK activation affects ERRα transcriptional activity and facilitates cellular energy balance. Additionally ERRα cross-talks with other nuclear receptors like PPAR (peroxisome proliferator-activated receptor) family members which are involved in lipid metabolism and glucose homeostasis.
- Associated diseases and disorders
ERRα has implications in metabolic conditions such as diabetes and obesity. The protein links to diabetes through its role in mitochondrial dysfunction and insulin resistance. Obesity-related studies point out how ERRα affects adaptive thermogenesis and lipid metabolism contributing to energy imbalance. ERRα-PPAR interactions highlight its association in metabolic disorders influencing various aspects of metabolism and disease progression. Understanding ERRα function offers therapeutic potential for targeting metabolic diseases.
Product promise
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3 product images
Western blot - Human ESRRA knockout MCF7 cell line (ab289292)
Western blot: Anti-ESRRA antibody (Anti-Estrogen Related Receptor alpha antibody ab137489) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Estrogen Related Receptor alpha antibody ab137489 was shown to bind specifically to ESRRA. A band was observed at 46 kDa in wild-type MCF7 cell lysates with no signal observed at this size in ESRRA knockout cell line. To generate this image, wild-type and ESRRA knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Estrogen Related Receptor alpha antibody (Anti-Estrogen Related Receptor alpha antibody ab137489) at 1/1000 dilution
Lane 1: Wild-type MCF7 cell lysate at 20 µg
Lanes 1 - 4: Western blot - Human ESRRA knockout MCF7 cell line (ab289292)
Lane 2: ESRRA knockout MCF7 cell lysate at 20 µg
Lane 3: Wild-type HAP1 cell lysate at 20 µg
Lane 4: ESRRA knockout HAP1 cell lysate at 20 µg
SecondaryLanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 46 kDa
Western blot - Human ESRRA knockout MCF7 cell line (ab289292)
Western blot: Anti-ESRRA antibody [EPR46Y] (Anti-Estrogen Related Receptor alpha antibody [EPR46Y] ab76228) staining at 1/2000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Estrogen Related Receptor alpha antibody [EPR46Y] ab76228 was shown to bind specifically to ESRRA. A band was observed at 46 kDa in wild-type MCF7 cell lysates with no signal observed at this size in ESRRA knockout cell line. To generate this image, wild-type and ESRRA knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Estrogen Related Receptor alpha antibody [EPR46Y] (Anti-Estrogen Related Receptor alpha antibody [EPR46Y] ab76228) at 1/2000 dilution
Lane 1: Wild-type MCF7 cell lysate at 20 µg
Lane 2: Western blot - Human ESRRA knockout MCF7 cell line (ab289292)
Lane 2: ESRRA knockout MCF7 cell lysate at 20 µg
Lane 3: Wild-type HAP1 cell lysate at 20 µg
Lane 4: ESRRA knockout HAP1 cell lysate at 20 µg
SecondaryLanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 46 kDa
Next Generation Sequencing - Human ESRRA knockout MCF7 cell line (ab289292)
40 bp deletion after Glu147 of the WT protein
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