Anti-Prohibitin antibody - Mitochondrial Marker

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Prohibitin antibody - Mitochondrial Marker (AB28172)

ab28172 staining Prohibitin in MCF7 cells. The cells were fixed with 100% methanol (5 min), 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-Triton for 1h. The cells were then incubated with the antibody ab28172 at 5μg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 μM for 1hour at room temperature.

• ICC/IF

Project

Immunocytochemistry/ Immunofluorescence - Anti-Prohibitin antibody - Mitochondrial Marker (AB28172)

ICC/IF image of ab28172 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab28172, 1μg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT^TM FX Signal Enhancer was used to quench autofluorescence. 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody - Mitochondrial Marker (AB28172)

IHC image of Prohibitin staining in human liver FFPE section, performed on a Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab28172, 5µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

• IHC-P

AbReview13651****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody - Mitochondrial Marker (AB28172)

ab28172 staining Prohibitin in mouse liver tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval in Citrate buffer pH 6.0 and blocking in 1.5% serum for 10 minutes. The primary antibody was diluted, 1/400 and incubated with sample for 1 hour. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary.

This image is courtesy of an anonymous Abreview

• WB

Project

Western blot - Anti-Prohibitin antibody - Mitochondrial Marker (AB28172)

All lanes:

Western blot - Anti-Prohibitin antibody - Mitochondrial Marker (ab28172) at 1 µg/mL

Lane 1:

Western blot - NIH/3T3 whole cell lysate (<a href='/en-us/products/cell-lysates/nih-3t3-whole-cell-lysate-ab7179'>ab7179</a>) at 10 µg

Lane 2:

MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Lane 3:

Brain (Mouse) Tissue Lysate at 10 µg

Lane 4:

Liver (Mouse) Tissue Lysate - normal tissue at 10 µg

Lane 5:

Heart (Mouse) Tissue Lysate at 10 µg

Lane 6:

Kidney (Mouse) Tissue Lysate at 10 µg

Lane 7:

Western blot - Mouse pancreas tissue lysate - total protein (<a href='/en-us/products/tissue-lysates/mouse-pancreas-tissue-lysate-total-protein-ab29363'>ab29363</a>) at 10 µg

Lane 8:

Testis (Mouse) Tissue Lysate - normal tissue at 10 µg

Lane 9:

Western blot - Mouse skeletal muscle tissue lysate - total protein (<a href='/en-us/products/tissue-lysates/mouse-skeletal-muscle-tissue-lysate-total-protein-ab29711'>ab29711</a>) at 10 µg

Lane 10:

Spinal Cord (Mouse) Tissue Lysate at 10 µg

Lane 11:

Ovary (Mouse) Tissue Lysate - normal tissue at 10 µg

Lane 12:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg

Lane 13:

Brain (Rat) Tissue Lysate - normal tissue at 10 µg

Lane 14:

Liver (Rat) Tissue Lysate at 10 µg

Lane 15:

Heart (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

• WB

Project

Western blot - Anti-Prohibitin antibody - Mitochondrial Marker (AB28172)

All lanes:

Western blot - Anti-Prohibitin antibody - Mitochondrial Marker (ab28172) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

Western blot - Jurkat whole cell lysate (<a href='/en-us/products/cell-lysates/jurkat-whole-cell-lysate-ab7899'>ab7899</a>) at 20 µg

Lane 3:

Western blot - A-431 whole cell lysate (<a href='/en-us/products/cell-lysates/a-431-whole-cell-lysate-ab7909'>ab7909</a>) at 20 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/10000 dilution

Predicted band size: 30 kDa

Observed band size: 30 kDa

false