NR3C1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2. The cells can still express the functionally active isoforms GRα -D1, D2, D3 from alternative start codons.
GCCR, GCRST, GCR_HUMAN, GR, Glucocorticoid receptor, Grl1, Nuclear receptor subfamily 3 group C member 1, glucocorticoid nuclear receptor variant 1, nr3c1, nuclear receptor subfamily 3, group C, member 1 (glucocorticoid receptor)
NR3C1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2. The cells can still express the functionally active isoforms GRα -D1, D2, D3 from alternative start codons.
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.
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.
The glucocorticoid receptor (GR) also known as the cortisol receptor is a type of nuclear receptor that functions as a transcription factor. This receptor has a molecular mass of approximately 97 kDa. GR is expressed in various tissues including the liver lung and immune cells. Its mechanical action involves binding to glucocorticoids—hormones like cortisol—which can regulate DNA transcription. When activated the receptor translocates into the cell nucleus where it can directly interact with DNA to regulate gene expression.
The glucocorticoid receptor plays a significant role in mediating the physiological effects of glucocorticoids. It controls processes like inflammation and immune response. The receptor is part of a larger receptor complex enhancing its effectiveness in gene regulation. It modulates expression levels of diverse genes involved in metabolism immune functionality and cellular growth. GR's regulatory actions make it contextual within various biological processes impacting cellular behavior extensively.
The glucocorticoid receptor integrates into significant signaling networks like the hypothalamic-pituitary-adrenal (HPA) axis and the inflammatory response pathway. This receptor coordinates with other proteins to control stress responses and inflammatory signals. In the HPA axis GR helps regulate cortisol levels and counteracts inflammatory cytokines. Its interaction with other receptors and transcription factors exemplifies its role in essential pathways that maintain homeostasis and stress adaptation within the organism.
Abnormal glucocorticoid receptor function links to conditions like Cushing's syndrome and glucocorticoid resistance. Cushing's syndrome characterized by excessive cortisol levels shows altered GR signaling. Similarly glucocorticoid resistance involves mutations or dysfunctions in GR that lead to improper hormone action affecting inflammation and immune responses. These diseases often involve other proteins such as various cytokines in the inflammatory response showing the broad impact of GR in disease processes.
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Anti-Glucocorticoid Receptor antibody [EPR19621] ab183127 Anti-Glucocorticoid Receptor antibody [EPR19621] was shown to specifically react with Glucocorticoid Receptor in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261766 (knockout cell lysate Human NR3C1 (Glucocorticoid Receptor) knockout HeLa cell lysate ab257009) was used. Wild-type and Glucocorticoid Receptor knockout samples were subjected to SDS-PAGE. Anti-Glucocorticoid Receptor antibody [EPR19621] ab183127 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866) were incubated overnight at 4° at 1 in 1000 dilution and 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-Glucocorticoid Receptor antibody [EPR19621] (Anti-Glucocorticoid Receptor antibody [EPR19621] ab183127) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: NR3C1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human NR3C1 (Glucocorticoid Receptor) knockout HeLa cell line (ab261766)
Lane 3: Wild-type A549 cell lysate at 20 µg
Lane 4: NR3C1 knockout A549 cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 85 kDa
Observed band size: 90-100 kDa
Homozygous: 1 bp insertion in exon 2.
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