RabMAb

Anti-Prohibitin抗体[EP2803Y] (ab75766)

概述

  • 产品名称Anti-Prohibitin抗体[EP2803Y]
    参阅全部 Prohibitin 一抗
  • 描述
    兔单克隆抗体[EP2803Y] to Prohibitin
  • 经测试应用适用于: ICC/IF, WB, IP, IHC-P, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Prohibitin (N terminal).

  • 阳性对照
    • MCF-7, A431, 293T and Ramos cell lysates; human kidney and hepatocellular carcinoma tissues. Jurkat (human acute T cell leukemia)
  • 常规说明

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

性能

应用

Our Abpromise guarantee covers the use of ab75766 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/100.
WB 1/10000 - 1/20000. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).
IP 1/40.
IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt 1/120.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Unpurified format: 1/1000

靶标

Anti-Prohibitin antibody [EP2803Y] 图像

  • All lanes : Anti-Prohibitin antibody [EP2803Y] (ab75766) at 1/20000 dilution

    Lane 1 : NIH/3T3 (mouse embryo) whole cell lysates
    Lane 2 : Mouse kidney tissue lysates
    Lane 3 : Rat brain tissue lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 30 kDa
    Additional bands at : 30 kDa. We are unsure as to the identity of these extra bands.
  • All lanes : Anti-Prohibitin antibody [EP2803Y] (ab75766) at 1/20000 dilution

    Lane 1 : MCF-7 (human breast carcinoma) whole cell lysates
    Lane 2 : A431 (human epidermoid carcinoma) whole cell lysates
    Lane 3 : HEK293 (human embryonic kidney) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 30 kDa
    Additional bands at : 30 kDa. We are unsure as to the identity of these extra bands.
  • All lanes : Anti-Prohibitin antibody [EP2803Y] (ab75766) at 1/20000 dilution

    Lane 1 : MCF-7 cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : 293T cell lysate
    Lane 4 : Ramos cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size : 30 kDa
    Observed band size : 30 kDa
  • ab75766 immunoprecipitating Prohibitin. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/40 and VeriBlot for IP secondary antibody (HRP) (ab131366) at a dilution of 1/10000.

    Lane 1: HEK293 (human embryonic kidney) whole cell lysate (10ug)
    Lane 2: HEK293 (human embryonic kidney) whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of  ab75766 in HEK293  (human embryonic kidney) whole cell lysate

  • ab75766 staining Prohibitin in rat kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody.

    Negative control 1: PBS in place of primary antibody.

  • ab75766 staining Prohibitin in mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody.

    Negative control 1: PBS in place of primary antibody.

  • ab75766 staining Prohibitin in human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody.

    Negative control 1: PBS in place of primary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labelling Prohibitin with ab75766 at 1/250.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse stomach tissue labelling Prohibitin with ab75766 at 1/250.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue labelling Prohibitin with ab75766 at 1/250.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue labelling Prohibitin with ab75766 at 1/250.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomach tissue labelling Prohibitin with ab75766 at 1/250.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal liver tissue labelling Prohibitin with ab75766.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal placenta tissue labelling Prohibitin with ab75766.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of breast carcinoma tissue labelling Prohibitin with ab75766.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling Prohibitin with ab75766 at 1/250.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular tissue labelling Prohibitin with ab75766 at 1/250.

  • ab75766 staining Prohibitin in NIH/3T3 (mouse embryonic fibroblast) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody. ab7291 (1/1000) and ab150120 (1/1000) were used as counterstains for primary antibody ab75766 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

    Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
    Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Prohibitin with ab75766 at 1/100. An Alexa Fluor® 488-conjugated anti-rabbit IgG was used as the secondary antibody.

  • Immunocytochemistry/Immunofluorescence analysis of NIH-3T3 cells labelling Prohibitin with ab75766 at 1/100. An Alexa Fluor® 488-conjugated anti-rabbit IgG was used as the secondary antibody.

  • Overlay histogram showing HepG2 cells stained with ab75766 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75766, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  • ab75766 staining Prohibitin in the human cell line A431 (human epidermoid carcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/120. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/500 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

Anti-Prohibitin antibody [EP2803Y] (ab75766)参考文献

ab75766 has not yet been referenced specifically in any publications.

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Thank you very much for your patience.

I have hear back now from the laboratories in regards to the potential crossreaction of the prohibitin antibodies with T. brucei. Unfortunately none of these antibodies seems to be likely to crossreact w...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Human Cell (keratinocytes)
Specification keratinocytes
Fixation Formaldehyde
Permeabilization No
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提交于 Jul 13 2011

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