Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)

Reviews (4)Q&A (4)References (19)

Overview

  • Product name
    Anti-Mineralocorticoid Receptor antibody [H10E4C9F]
    See all Mineralocorticoid Receptor primary antibodies
  • Description
    Mouse monoclonal [H10E4C9F] to Mineralocorticoid Receptor
  • Host species
    Mouse
  • Specificity
    Detects mineralocorticoid receptor (MR).
  • Tested applications
    Suitable for: Flow Cyt, Inhibition Assay, ELISA, IHC-P, WB, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Rabbit, Chicken, Hamster, Cow, Dog, Human
  • Immunogen

    Aldosterone 3. This antibody was produced using the anti idiotypic method.

Properties

Applications

Our Abpromise guarantee covers the use of ab2774 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/20. PubMed: 20573992

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 
Inhibition Assay Use at an assay dependent concentration.

ab2774 blocks the binding of aldosterone since it is directed against the steroid binding domain of the mineralocorticoid receptor.
ELISA Use at an assay dependent concentration.
IHC-P 1/100. Immunohistochemical staining of MR in rabbit atrium with ab2774 results in strong staining of myocytes and endothelial cells. In immunohistochemical studies, staining with ab2774 is blocked by pre incubating the sample with aldosterone.
WB 1/250. Predicted molecular weight: 107 kDa. By Western blot, this antibody detects a 116 kDa protein representing MR in rabbit heart homogenate. Due to the extremely low levels of MR expressed in native tissues, it is recommended that enhanced detection systems be used for Western blotting such as enhanced chemiluminescence.
ICC/IF 1/100.
  • Application notes
    Is unsuitable for IP.

    • Target

    • Function
      Receptor 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 specificity
      Ubiquitous. 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 disease
      Defects 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 similarities
      Belongs to the nuclear hormone receptor family. NR3 subfamily.
      Contains 1 nuclear receptor DNA-binding domain.
    • Domain
      Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain.
    • Post-translational
      modifications
      Phosphorylated.
    • Cellular localization
      Cytoplasm. 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.
    • Information by UniProt
    • Database links
      see all
    • Alternative names
      • Aldosterone receptor antibody
      • MCR antibody
      • MCR_HUMAN antibody
      • MGC133092 antibody
      • Mineralocorticoid receptor antibody
      • MLR antibody
      • MR antibody
      • NR3 C2 antibody
      • NR3C2 antibody
      • NR3C2 protein antibody
      • Nuclear receptor subfamily 3 group C member 2 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)Image courtesy of an abreview from Mrs. Barbara Heitkönig.

      Formalin-fixed, paraffin-embedded cow kidney tissue stained for Mineralocorticoid Receptor using ab2774 at 1/200 dilution in immunohistochemical analysis.

      Heat mediated Antigen retrieval using Citrate buffer, 10 mM pH 6.0 was used.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      ab2774 (4µg/ml) staining mineralocorticoid receptor in human pancreas, using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of the islets of Langerhans and some weaker staining of the exocrine cells of the pancreas.
      Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
    • Immunocytochemistry/ Immunofluorescence - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunocytochemistry/ Immunofluorescence - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)Image courtesy of an anonymous Abreview.
      ab2774 staining Mineralocorticoid Receptor in hamster CHO K1 cells by Immunocytochemistry/ Immunofluorescence. The cells were formaldehyde fixed and then blocked using TBS, 3% Dried Milk, 0.1 % Triton X-100 for 20 minutes at 22°C. Samples were then incubated with primary antibody at 1/100 for 1 hour at 22°C. The secondary antibody used was a goat anti-mouse IgG conjugated to FITC used at a 1/400 dilution (left hand image). In the right hand image DAPI was used to stain the cell nuclei blue.
    • Flow Cytometry - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Flow Cytometry - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Overlay histogram showing HEK293 cells stained with ab2774 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2774, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human colon carcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing Mineralocorticoid Receptor ab2774 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing Mineralocorticoid Receptor ab2774 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human kidney tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing Mineralocorticoid Receptor ab2774 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

    • Immunocytochemistry/ Immunofluorescence - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunocytochemistry/ Immunofluorescence - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunofluorescent analysis of Mineralocorticoid Receptor using Mineralocorticoid Receptor Monoclonal antibody (H10E4C9F) ab2774 shows staining in HEK293 cells. Mineralocorticoid Receptor staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Mineralocorticoid Receptor ab2774 at a dilution of 1:20-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

    • Immunocytochemistry/ Immunofluorescence - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunocytochemistry/ Immunofluorescence - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunofluorescent analysis of Mineralocorticoid Receptor using Mineralocorticoid Receptor Monoclonal antibody (H10E4C9F) ab2774 shows staining in HeLa cells. Mineralocorticoid Receptor staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Mineralocorticoid Receptor ab2774 at a dilution of 1:20-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

    • Immunocytochemistry/ Immunofluorescence - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunocytochemistry/ Immunofluorescence - Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)
      Immunofluorescent analysis of Mineralocorticoid Receptor using Mineralocorticoid Receptor Monoclonal antibody (H10E4C9F) ab2774 shows staining in HepG2 cells. Mineralocorticoid Receptor staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Mineralocorticoid Receptor ab2774 at a dilution of 1:20-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody . Images were taken at 60X magnification.

    References

    This product has been referenced in:
    • Tata CM  et al. Antihypertensive effects of the hydro-ethanol extract of Senecio serratuloides DC in rats. BMC Complement Altern Med 19:52 (2019). Read more (PubMed: 30819180) »
    • Wang J  et al. Aldosterone induced up-expression of ICAM-1 and ET-1 in pancreatic islet endothelium may associate with progression of T2D. Biochem Biophys Res Commun 512:750-757 (2019). Read more (PubMed: 30926169) »
    See all 19 Publications for this product

    Publishing research using ab2774? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-8 of 8 Abreviews or Q&A

    Question

    I am using this antibody in WB of mouse brain tissue lysates and getting no bands. I have loaded 10-60ug of total protein. The membrane is blocked with either BSA or milk. I have tested antibody concentrations 1:1000 - 1:200. Can I try another antibody to this target?

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    Answer

    Thank you for contacting Abcam regarding ab2774.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.
    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

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    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Mineralocorticoid Receptor antibody [H10E4C9F]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Cow Tissue sections (Kidney)
    Specification
    Kidney
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citratbuffer, 10 mM pH 6,0
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 22°C
    Read More

    Mrs. Barbara Heitkönig

    Verified customer

    Submitted Jul 07 2011

    Question

    Our lab recently purchased three antibodies (Nitrotyrosine, Mineralocorticoid Receptor and TNF-a) from Abcam. We have stored them at our refrigerator on getting them without checking them out. Today I took them out to make aliquots and noticed the two (MR: ab2774 and TNF-a: ab6671) of them are empty.   I do not know what happened on them. As you know they are very expensive. Please let me know what I can do for them.  

    Read More
    Answer

    Thank you for contacting us and letting us know about this problem It is possible that the water evaporated from the vials, as both of these antibodies were supplied in small volumes and sometimes the water does evaporate. Ab2774 should contain 100µl of liquid, and ab6771 should contain 40µl. I recommend first spinning the vials down to make sure there is no liquid in the cap. Then, add deionized water until the vials contain the appropriate volumes. If you do have any problems with the functionality of these antibodies, please let me know and I will be happy to send you replacement vials.

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    Western blot abreview for Anti-Mineralocorticoid Receptor antibody [H10E4C9F]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (kidney fibroblast)
    Loading amount
    3 µg
    Specification
    kidney fibroblast
    Treatment
    aldosterone 5nM 20nM
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More

    Dong Chen

    Verified customer

    Submitted Sep 02 2009

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Mineralocorticoid Receptor antibody [H10E4C9F]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Hamster Cell (Expression of Human Mineralocorticoid receptor in)
    Specification
    Expression of Human Mineralocorticoid receptor in
    Fixative
    Formaldehyde
    Blocking step
    TBS + 3% Dried Milk + 0.1 % Triton X-100 as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 29 2008

    Western blot abreview for Anti-Mineralocorticoid Receptor antibody [H10E4C9F]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Dog Tissue lysate - whole (left ventricle)
    Loading amount
    90 µg
    Specification
    left ventricle
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. sudhish mishra

    Verified customer

    Submitted Jun 19 2007

    Question

    I did not order this antibody, I was given an aliquot. The Ab catalog # is ab2774 (H10E4C9G, Mineralcorticoid Receptor) and I would like to know the concentration of the Ab in the 100ul vial that was originally purchased. The data sheet does not contain this information. I do not need to know the concentration of a starting dilution, just the concentration of the Ab itself. For example: 10ug/100ul. I have already contacted Abcam via phone and they did not have access to this information.

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    Answer

    Thank you for your enquiry. As the purity of ab2774 is ascites, and is therefore unpurified antibody, we cannot provide a concentration. Please contact us again if you have any additional questions.

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    Question

    Some days ago I ordered an antibody from your company. I have recieved the package today. Unfortunatly, the tube you sent me does not contain any liquid material, however, according to the data sheet it should be formulated as 100 microliter liquid, (ascities diluted in PBS). The antibody ordered is the mouse monoclonal [H10E4C9F] one against the mineralocorticoid receptor (ab2774; LOT 58328). Please contact me as soon as possible in order to try to solve this problem.

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    Answer

    Dear Zoltan, The Mineralocorticoid Receptor ab2774 antibody should have been in liquid form. We do lyophilize some of our products to protect the activity of the antibody during shipment; however we do not show this antibody being lyophilized. Therefore, we will send you a replacement of the ab2774 free of charge. Please let me know if you have any other concerns about any of our quality guaranteed antibodies. Best Wishes,

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