Anti-Prohibitin antibody - Mitochondrial Marker (ab28172)
- Datasheet
- Specific References (38)
- Protocols
Overview
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Product nameAnti-Prohibitin antibody - Mitochondrial Marker
See all Prohibitin primary antibodies -
DescriptionRabbit polyclonal to Prohibitin - Mitochondrial Marker
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Host speciesRabbit
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Tested applicationsSuitable for: WB, ICC/IF, IHC-Pmore details
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Species reactivityReacts with: Mouse, Rat, Cow, Dog, Human, Pig
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human Prohibitin.
(Peptide available as ab28492.)
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Positive control
- ab28172 antibody gave a positive signal in the following Whole Cell Lysates: HeLa, A431, Jurkat, NIH 3T3, MEF-1 and PC12. In addition, ab28172 antibody gave a positive signal in the following Tissue Lysates: Brain (Mouse), Liver (Mouse), Heart (Mouse), Kidney (Mouse), Pancreas (Mouse), Testis (Mouse), Skeletal Muscle (Mouse), Spinal Cord (Mouse), Ovary (Mouse), Brain (Rat), Liver (Rat) and Heart (Rat).
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Positive Controls
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Related Products
Applications
Our Abpromise guarantee covers the use of ab28172 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa). | |
| ICC/IF | Use a concentration of 5 µg/ml. | |
| IHC-P | Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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FunctionProhibitin inhibits DNA synthesis. It has a role in regulating proliferation. As yet it is unclear if the protein or the mRNA exhibits this effect. May play a role in regulating mitochondrial respiration activity and in aging.
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Tissue specificityWidely expressed in different tissues.
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Sequence similaritiesBelongs to the prohibitin family.
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Developmental stageLevels of expression in fibroblasts decrease heterogeneously during cellular aging.
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Cellular localizationMitochondrion inner membrane.
- Information by UniProt
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Database links
- Entrez Gene: 530409 Cow
- Entrez Gene: 480547 Dog
- Entrez Gene: 5245 Human
- Entrez Gene: 18673 Mouse
- Entrez Gene: 25344 Rat
- Omim: 176705 Human
- SwissProt: Q3T165 Cow
- SwissProt: P35232 Human
see all -
Alternative names
- Epididymis luminal protein 215 antibody
- Epididymis secretory sperm binding protein Li 54e antibody
- HEL 215 antibody
see all
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 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. -
Western blot - Anti-Prohibitin antibody - Mitochondrial Marker (ab28172)All lanes : Anti-Prohibitin antibody - Mitochondrial Marker (ab28172) at 1 µg/ml
Lane 1 :NIH 3T3 whole cell lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : Brain (Mouse) Tissue Lysate
Lane 4 : Liver (Mouse) Tissue Lysate - normal tissue
Lane 5 : Heart (Mouse) Tissue Lysate
Lane 6 : Kidney (Mouse) Tissue Lysate
Lane 7 :Mouse pancreas tissue lysate - total protein (ab29363)
Lane 8 : Testis (Mouse) Tissue Lysate - normal tissue
Lane 9 :Mouse skeletal muscle tissue lysate - total protein (ab29711)
Lane 10 : Spinal Cord (Mouse) Tissue Lysate
Lane 11 : Ovary (Mouse) Tissue Lysate - normal tissue
Lane 12 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 13 : Brain (Rat) Tissue Lysate - normal tissue
Lane 14 : Liver (Rat) Tissue Lysate
Lane 15 : Heart (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
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 -
Immunocytochemistry/ Immunofluorescence - Anti-Prohibitin antibody - Mitochondrial Marker (ab28172)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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody - Mitochondrial Marker (ab28172)This image is courtesy of an anonymous Abreviewab28172 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 : Anti-Prohibitin antibody - Mitochondrial Marker (ab28172) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 :Jurkat whole cell lysate (ab7899)
Lane 3 :A431 whole cell lysate (ab7909)
Lysates/proteins at 20 µg per lane.
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 -
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-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).
Protocols
References
This product has been referenced in:
- Festa BP et al. Impaired autophagy bridges lysosomal storage disease and epithelial dysfunction in the kidney. Nat Commun 9:161 (2018). Read more (PubMed: 29323117) »
- Ramseyer VD et al. Vacuolar protein sorting 13C is a novel lipid droplet protein that inhibits lipolysis in brown adipocytes. Mol Metab 7:57-70 (2018). Read more (PubMed: 29175050) »
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