Anti-Prohibitin antibody - Mitochondrial Marker (ab28172)

Overview

  • Product nameAnti-Prohibitin antibody - Mitochondrial Marker
    See all Prohibitin primary antibodies
  • Description
    Rabbit polyclonal to Prohibitin - Mitochondrial Marker
  • Tested applicationsSuitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Dog, Human, Pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human Prohibitin.

    (Peptide available as ab28492.)

  • 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

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

Anti-Prohibitin antibody - Mitochondrial Marker images

  • 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
    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
  • 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
    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
  • 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.

  • 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).

References for Anti-Prohibitin antibody - Mitochondrial Marker (ab28172)

This product has been referenced in:
  • Helenius TO  et al. Keratin 8 absence down-regulates colonocyte HMGCS2 and modulates colonic ketogenesis and energy metabolism. Mol Biol Cell 26:2298-310 (2015). Read more (PubMed: 25904331) »
  • Chakrabortty SK  et al. Extracellular vesicle-mediated transfer of processed and functional RNY5 RNA. RNA 21:1966-79 (2015). WB ; Human . Read more (PubMed: 26392588) »

See all 23 Publications for this product

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (C2C12 cell line)
Specification C2C12 cell line
Fixative Paraformaldehyde
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
Username

Abcam user community

Verified customer

Submitted Jan 25 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (pancreatic)
Specification pancreatic
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: DAKO Antigen retrieval buffer pH6
Permeabilization No
Username

Dr. Ilka Werner-Martini

Verified customer

Submitted Oct 15 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (breast cancer)
Specification breast cancer
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Dako Target Retrieval Solution
Permeabilization No
Blocking step 5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C
Username

Mr. Antibody Solutions

Verified customer

Submitted May 26 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (liver)
Specification liver
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
Permeabilization No
Blocking step Serum as blocking agent for 10 minute(s) · Concentration: 1.5%
Username

Abcam user community

Verified customer

Submitted Mar 11 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Username

Dr. Kirk McManus

Verified customer

Submitted Oct 03 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"