Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-RPE65 antibody [EPR22579-44] (ab231782)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22579-44] to RPE65
  • Suitable for: IHC-P, WB, IHC-Fr, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-RPE65 antibody [EPR22579-44]
    See all RPE65 primary antibodies

  • Description

    Rabbit monoclonal [EPR22579-44] to RPE65

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IHC-Fr
    Mouse
    IHC-P
    Mouse
    Rat
    Human
    IP
    Mouse
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human eye tissue lysate; mouse eye tisue lysate; rat eye tissue lysate. IHC-P: Human retina tissue; mouse retina tissue; rat retina tissue. IHC-Fr: Mouse retina tissue; rat retina tissue. IP: Mouse eyeball lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPE65 antibody [EPR22579-44] (ab231782)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPE65 antibody [EPR22579-44] (ab231782)

    Immunohistochemical analysis of paraffin-embedded mouse retina tissue labeling RPE65 with ab231782 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on pigment epithelium cells of mouse retina (PMID: 25941382, PMID: 22238088 is observed. Counter stained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPE65 antibody [EPR22579-44] (ab231782)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPE65 antibody [EPR22579-44] (ab231782)

    Immunohistochemical analysis of frozen section of 4%PFA-fixed, 0.2% Triton X-100 permeabilized rat retinal tissue labeling RPE65 with ab231782 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution (green). Positive staining on retinal pigment epithelium (PMID: 17848510) is observed. The nuclear counter stain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

  • Immunoprecipitation - Anti-RPE65 antibody [EPR22579-44] (ab231782)
    Immunoprecipitation - Anti-RPE65 antibody [EPR22579-44] (ab231782)

    RPE65 was immunoprecipitated from 0.35 mg of mouse eyeball lysate with ab231782 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab231782 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

    Lane 1: Mouse eyeball lysate 10 μg (Input).
    Lane 2: ab231782 IP in mouse eyeball lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab231782 in mouse eyeball lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 minutes.

  • Western blot - Anti-RPE65 antibody [EPR22579-44] (ab231782)
    Western blot - Anti-RPE65 antibody [EPR22579-44] (ab231782)
    All lanes : Anti-RPE65 antibody [EPR22579-44] (ab231782) at 1/1000 dilution

    Lane 1 : Human eye tissue lysate
    Lane 2 : Mouse eye tissue lysate
    Lane 3 : Rat eye tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 61 kDa
    Observed band size: 65 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure times.

    Lane 1: 26 seconds. Lanes 2 & 3: 3 minutes.

    The molecular weight observed is consistent with what has been described in the literature (PMID:17848510).

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPE65 antibody [EPR22579-44] (ab231782)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPE65 antibody [EPR22579-44] (ab231782)

    Immunohistochemical analysis of paraffin-embedded human retina tissue labeling RPE65 with ab231782 at 1/4000 dilution, followed by ready to use AP-labeled secondary antibody kit. Cytoplasmic staining on pigment epithelium cells of human retina (PMID: 25941382; PMID: 22238088) is observed. Counter stained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use AP-labeled secondary antibody kit.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    The section was incubated with ab231782 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Immunohistochemistry (Frozen sections) - Anti-RPE65 antibody [EPR22579-44] (ab231782)
    Immunohistochemistry (Frozen sections) - Anti-RPE65 antibody [EPR22579-44] (ab231782)

    Immunohistochemical analysis of frozen section of 4%PFA-fixed, 0.2% Triton X-100 permeabilized mouse retinal tissue labeling RPE65 with ab231782 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution (green). Positive staining on retinal pigment epithelium (PMID: 17848510) is observed. The nuclear counter stain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPE65 antibody [EPR22579-44] (ab231782)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPE65 antibody [EPR22579-44] (ab231782)

    Immunohistochemical analysis of paraffin-embedded rat retina tissue labeling RPE65 with ab231782 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on pigment epithelium cells of rat retina (PMID: 25941382, PMID: 22238088) is observed. Counter stained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

     

  • Anti-RPE65 antibody [EPR22579-44] (ab231782)
    Anti-RPE65 antibody [EPR22579-44] (ab231782)

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