Anti-Prohibitin antibody - Mitochondrial Marker

• Supplier: Abcam
• Catalog: ab28172
• Prices: $400.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: WB ICC/IF ICC/IF IHC-P
• Species Reactivities: Mouse, Rat, Bovine, Dog, Human, Pig
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human Prohibitin
• Description: Rabbit Polyclonal
• Gene: PHB
• Conjugate: Unconjugated

Product images

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-iTTM 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).

All lanes : Anti-Prohibitin antibody - Mitochondrial Marker (ab28172) at 1 µg/mlLane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 3 : Brain (Mouse) Tissue LysateLane 4 : Liver (Mouse) Tissue Lysate - normal tissueLane 5 : Heart (Mouse) Tissue LysateLane 6 : Kidney (Mouse) Tissue LysateLane 7 : Pancreas (Mouse) Tissue Lysate (ab29363)Lane 8 : Testis (Mouse) Tissue Lysate - normal tissueLane 9 : Skeletal Muscle (Mouse) Tissue Lysate (ab29711)Lane 10 : Spinal Cord (Mouse) Tissue LysateLane 11 : Ovary (Mouse) Tissue Lysate - normal tissueLane 12 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell LysateLane 13 : Brain (Rat) Tissue Lysate - normal tissueLane 14 : Liver (Rat) Tissue LysateLane 15 : Heart (Rat) Tissue LysateLysates/proteins at 10 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilutionPerformed under reducing conditions.

All lanes : Anti-Prohibitin antibody - Mitochondrial Marker (ab28172) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)Lysates/proteins at 20 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/10000 dilutionPerformed under reducing conditions.

IHC image of Prohibitin staining in human liver FFPE section, performed on a BondTM 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.

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.See Abreview

ICC/IF image of ab28172 stained MCF7 cells. The cells were 100% methanol fixed (5 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 (ab28172, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 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 1.43µM.

Product References

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