RXRA KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
FLJ00280, FLJ00318, FLJ16020, FLJ16733, MGC102720, NR2B1, Nuclear receptor subfamily 2 group B member 1, OTTHUMP00000022510, RXR alpha1, RXRA_HUMAN, Retinoic acid receptor RXR-alpha, Retinoid X nuclear receptor alpha, Retinoid X receptor alpha
RXRA KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
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.
Retinoid X Receptor alpha (RXRA) also referred to as NR2B1 is a nuclear receptor with a mass of approximately 55 kDa. RXRA functions as a transcription factor and plays a significant role in regulating gene expression by forming heterodimers with other nuclear receptors. RXRA is expressed in many tissues with notable levels in organs like the liver lungs and kidneys. Its ability to bind DNA sequences known as retinoic acid response elements allows RXRA to regulate diverse physiological processes.
RXRA serves an important role in metabolic regulation especially in lipid metabolism and glucose homeostasis. It forms heterodimers with partners such as peroxisome proliferator-activated receptors (PPARs) enhancing its capacity to modulate various metabolic pathways. These heterodimers facilitate the transcription of genes that respond to nutritional changes. RXRA also influences immune responses and cell differentiation making it important for maintaining cellular health.
RXRA operates within the retinoid signaling and lipid metabolism pathways. Through its interaction with PPARs particularly PPAR-γ and PPAR-α RXRA becomes an integral part of the signaling that controls fatty acid storage and glucose metabolism. These pathways influence cellular function and energy balance impacting the body's overall metabolic status.
RXRA has associations with metabolic diseases such as diabetes and cardiovascular disorders. Dysregulation of RXRA expression or function can disrupt lipid and glucose metabolism leading to insulin resistance and metabolic syndrome. The RXRA-PPAR-γ interaction is essential in these conditions with the pair influencing adipogenesis and inflammatory responses. Understanding and targeting RXRA interactions can provide therapeutic prospects for these metabolic disorders.
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Terms & Conditions.
Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] ab125001 was shown to react with Anti-Retinoid X Receptor alpha in wild-type HCT 116 cells in western blot with loss of signal observed in RXRA knockout cell line ab273708 (RXRA knockout cell lysate Human RXRA knockout HCT116 cell lysate ab275245). Wild-type and RXRA knockout HCT 116 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] ab125001 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] ab125001) at 1/1000 dilution
Lane 1: Wild-type HCT116 cell lysate at 20 µg
Lane 2: RXRA knockout HCT116 cell lysate at 20 µg
Lane 2: Western blot - Human RXRA knockout HCT116 cell line (ab273708)
Lane 3: HeLa cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 51 kDa
Observed band size: 53 kDa
Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] ab125001 staining Retinoid X Receptor alpha in wild-type HCT116 cells (top panel) and RXRA knockout HCT116 cells (bottom panel) (ab273708). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] ab125001 at 0.2μg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Allele-1: 1 bp insertion in exon 1.
All lanes: Western blot - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] ab125001) at 1/500 dilution
Lane 1: Wild-type HCT116 lysate at 20 µg
Lane 2: Western blot - Human RXRA knockout HCT116 cell line (ab273708) at 20 µg
Observed band size: 51 kDa
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