製品の概要

  • 製品名

    Anti-Prohibitin antibody - Mitochondrial Marker
    Prohibitin 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to Prohibitin - Mitochondrial Marker
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: WB, ICC/IF, IHC-Pmore details
  • 種交差性

    交差種: Mouse, Rat, Cow, Dog, Human, Pig
  • 免疫原

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

    Immunogen の所有権に関して (Peptide available as ab28492.)

  • ポジティブ・コントロール

    • Recombinant Human Prohibitin protein (ab114204) can be used as a positive control in WB. 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).

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

画像

  • 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.
  • 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

  • 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.

  • 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

  • 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

  • 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).

参考文献

This product has been referenced in:

  • Hensen F  et al. Mitochondrial RNA granules are critically dependent on mtDNA replication factors Twinkle and mtSSB. Nucleic Acids Res N/A:N/A (2019). Read more (PubMed: 30715486) »
  • Huang C  et al. FoxM1 Induced Paclitaxel Resistance via Activation of the FoxM1/PHB1/RAF-MEK-ERK Pathway and Enhancement of the ABCA2 Transporter. Mol Ther Oncolytics 14:196-212 (2019). Read more (PubMed: 31334335) »
See all 46 Publications for this product

レビューと Q&A

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1-5 of 5 Abreviews

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

Abcam user community

Verified customer

投稿 Jan 25 2011

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

Dr. Ilka Werner-Martini

Verified customer

投稿 Oct 15 2010

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

Mr. Antibody Solutions

Verified customer

投稿 May 26 2009

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%

Abcam user community

Verified customer

投稿 Mar 11 2009

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

投稿 Oct 03 2007

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