Anti-Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-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 - Mineralocorticoid Receptor antibody [H10E4C9F] (ab2774)

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 22ºC. Samples were then incubated with primary antibody at 1/100 for 1 hour at 22ºC. 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.

Image courtesy of an anonymous Abreview.

Flow Cytometry-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 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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.