Anti-Prohibitin antibody - Mitochondrial Marker

6 product images

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  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.

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  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 (NIH/3T3 whole cell lysate ab7179) 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 (Mouse pancreas tissue lysate - total protein ab29363) 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 (Mouse skeletal muscle tissue lysate - total protein ab29711) 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

  Performed under reducing conditions.

  Predicted band size: 30 kDa

  Observed band size: 30 kDa

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  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 Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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.

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  This image is courtesy of an anonymous customer review

  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.
  

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  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 (Jurkat whole cell lysate ab7899) at 20 µg

  Lane 3: Western blot - A-431 whole cell lysate (A-431 whole cell lysate ab7909) at 20 µg

  Secondary

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

  Performed under reducing conditions.

  Predicted band size: 30 kDa

  Observed band size: 30 kDa

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  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).