Mouse monoclonal [H3122] to Mineralocorticoid Receptor
SpecificityIn immunohistochemistry this antibody gives good staining in colon and kidney tissue. However, it has not been possible to detect endogenous expression by Western; as positive control a mouse cell line transfected with cDNA was used for this application.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 0.2 µg/ml.
Use at an assay dependent concentration.
1/20 - 1/40.
Use at an assay dependent concentration.
Use a concentration of 1 µg/ml. Predicted molecular weight: 107 kDa. Block with 5% milk and incubate primary antibody in blocking buffer.
Use at an assay dependent concentration. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
Use a concentration of 1 µg/ml.
FunctionReceptor for both mineralocorticoids (MC) such as aldosterone and glucocorticoids (GC) such as corticosterone or cortisol. Binds to mineralocorticoid response elements (MRE) and transactivates target genes. The effect of MC is to increase ion and water transport and thus raise extracellular fluid volume and blood pressure and lower potassium levels.
Tissue specificityUbiquitous. Highly expressed in distal tubules, convoluted tubules and cortical collecting duct in kidney, and in sweat glands. Detected at lower levels in cardiomyocytes, in epidermis and in colon enterocytes.
Involvement in diseaseDefects in NR3C2 are a cause of autosomal dominant pseudohypoaldosteronism type I (AD-PHA1) [MIM:177735]. PHA1 is characterized by urinary salt wasting, resulting from target organ unresponsiveness to mineralocorticoids. There are 2 forms of PHA1: the autosomal dominant form that is mild, and the recessive form which is more severe and due to defects in any of the epithelial sodium channel subunits. In AD-PHA1 the target organ defect is confined to kidney. Clinical expression can vary from asymptomatic to moderate. It may be severe at birth, but symptoms remit with age. Familial and sporadic cases have been reported. Defects in NR3C2 are a cause of early-onset hypertension with severe exacerbation in pregnancy (EOHSEP) [MIM:605115]. Inheritance is autosomal dominant. The disease is characterized by the onset of severe hypertension before the age of 20, and by suppression of aldosterone secretion.
Sequence similaritiesBelongs to the nuclear hormone receptor family. NR3 subfamily. Contains 1 nuclear receptor DNA-binding domain.
DomainComposed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain.
Cellular localizationCytoplasm. Nucleus. Endoplasmic reticulum membrane. Cytoplasmic and nuclear in the absence of ligand; nuclear after ligand-binding. When bound to HSD11B2, it is found associated with the endoplasmic reticulum membrane.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Mineralocorticoid Receptor [H3122] antibody (ab41912)This image is courtesy of an Abreview submitted by Michael Hasstedt
ab41912 staining Mineralocorticoid in male European Starling brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were transcardially perfused with paraformaldehyde, and immersed in paraformaldyhyde for 24 hours and then 30% sucrose for 24 hours. Slices were cut at a 40 micron thickness. Tissue samples were permeablized with Triton and blocked with 10% serum for 1 hour at 21°C. The sample was incubated with primary antibody (1/500) for 24 hours. A biotin-conjugated Goat anti-mouse IgG monoclonal (1/400) was used as the secondary antibody.
ICC/IF image of ab41912 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab41912, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Flow Cytometry - Mineralocorticoid Receptor antibody [H3122] (ab41912)Image courtesy of Luciana Besedovsky by Abreview.
ab41912 used in Flow Cytometry. Human peripheral blood leukocyte cells were fixed with paraformaldehyde, permeabilized with saponin and then stained with ab41912 (which had been directly conjugated to Alexa Fluor 647), used at a 1/50 dilution. Gated on CD3+ lymphocytes (T cells).