Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Prohibitin antibody [II-14-10] - Mitochondrial Marker (ab1836)

Formalin fixed paraffin embedded human heart stained with Prohibitin antibody ab1836.

Immunohistochemistry (Formalin-fixed paraffin-embedded sections) - Prohibitin antibody [II-14-10] - Mitochondrial Marker (ab1836)

ab1836 at a 1/100 dilution staining mouse Cardiac muscle tissue sections by Immunohistochemistry (Formalin-fixed paraffin-embedded sections). Following heat mediated antigen retrieval the antibody was incubated with the tissue for 12 hours. Bound antibody was detected with a HRP conjuaged anti-mouse antibody.

The picture shown is a transverse section of mouse ventricular wall; from TEM studies the distribution of mitochondria among the myofibrils is well-established, and so in this 2-um-thick section there exists both minimal superposition as well as a staining pattern which is consistent with what is expected for cardiac mitochondrial patterns (arrows in the inset indicate groups of prohibitin-positive mitochondria).

This image is courtesy of an Abreview submitted by Mike Forbes on 29 March 2006.

Immunocytochemistry/ Immunofluorescence - Prohibitin antibody [II-14-10] - Mitochondrial Marker (ab1836)

ICC/IF image of ab1836 stained HepG2 cells. The cells were 4% PFA fixed (10 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 (ab1836, 1æg/ml) overnight at +4øC. The secondary antibody (green)ÿwas Alexa Fluor© 488 goat anti- 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 1.43æM.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Prohibitin antibody [II-14-10] - Mitochondrial Marker (ab1836)

ab1836 staining Prohibitin in normal human skin tissue sections by IHC-P (formaldehyde-fixed paraffin-embedded sections). Tissue samples were fixed with formaldehyde and blocked with 10% serum for 30 minutes at 21ºC; antigen retrival was by heat mediation in Citric buffer (pH6). The sample was incubated with primary antibody (1/200 in TBS + 1% BSA) at 21ºC for 1 hour. An undiluted HRP-conjugated Goat polyclonal to mouse IgG was used as secondary antibody.

This image is courtesy of an anonymous Abreview

Flow Cytometry-Prohibitin antibody [II-14-10] - Mitochondrial Marker(ab1836)

Overlay histogram showing HeLA cells stained with ab1836 (red line). The cells were fixed with 100% 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 (ab1836, 1µg/1x10⁶ cells) 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.