Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22227-6] to LINE-1 ORF1p
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-LINE-1 ORF1p antibody [EPR22227-6]
    See all LINE-1 ORF1p primary antibodies

  • Description

    Rabbit monoclonal [EPR22227-6] to LINE-1 ORF1p

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: NCCIT, MCF7 and K562 whole cell lysate. IHC-P: Human pancreatic cancer, colon cancer and lung cancer tissue. ICC/IF: NCCIT and MCF7 cells. Flow cyt: NCCIT cells. IP: NCCIT whole cell 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

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR22227-6

  • Isotype

    IgG

  • Research areas

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)
    Immunocytochemistry/ Immunofluorescence - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NCCIT (human pluripotent embryonic carcinoma epithelial cell) cells labeling LINE-1 ORF1p with ab245249 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic with punctate foci and nucleolar staining in NCCIT cells (PMID: 17562864; 15028673). The nuclear counter stain is DAPI (blue).
    Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
    Secondary antibody only control:  Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)
    Immunocytochemistry/ Immunofluorescence - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labeling LINE-1 ORF1p with ab245249 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic with punctate foci and nucleolar staining in MCF7 cells (PMID: 15028673; 17562864). The nuclear counter stain is DAPI (blue).
    Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
    Secondary antibody only control:  Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution..

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)

    Immunohistochemical analysis of paraffin-embedded human colon cancer (Panel A) and adjacent normal tissue (Panel B) labeling LINE-1 ORF1p with ab245249 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in colon cancer (Panel A) but negative in adjacent normal tissue (Panel B). (PMID:24607009). Counterstained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)

    Immunohistochemical analysis of paraffin-embedded human pancreatic cancer (Panel A) and adjacent normal tissue (Panel B) labeling LINE-1 ORF1p with ab245249 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in pancreatic cancer (Panel A) but negative in adjacent normal tissue (Panel B). (PMID:24607009). Counterstained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)

    Immunohistochemical analysis of paraffin-embedded human lung cancer (Panel A) and adjacent normal tissue (Panel B) labeling LINE-1 ORF1p with ab245249 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in lung cancer (Panel A) but negative in adjacent normal tissue (Panel B). (PMID:24607009). Counterstained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

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

  • Immunoprecipitation - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)
    Immunoprecipitation - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)

    LINE-1 ORF1p was immunoprecipitated from 0.35 mg NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate with ab245249 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab245249 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

    Lane 1: NCCIT whole cell lysate 10 µg (Input).
    Lane 2: ab245249 IP in NCCIT whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab245249 in NCCIT whole cell lysate.

    The lower 33-kDa band is likely to be the truncated form (PMID: 29505568).

    Blocking/Dilution buffer: 5% NFDM/TBST.
    Exposure time: 1 second.

  • Flow Cytometry - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)
    Flow Cytometry - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HCT 116 (human colorectal carcinoma epithelial cell, Left panel) / NCCIT (human pluripotent embryonic carcinoma epithelial cell, Right panel) cell lines labeling LINE-1 ORF1p with ab245249 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

    The expression level on HCT 116 (low) and NCCIT (high) is consistent with literature (PMID: 27016617).

  • Western blot - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)
    Western blot - Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)
    All lanes : Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249) at 1/1000 dilution

    Lane 1 : NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate
    Lane 2 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
    Lane 3 : K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
    Lane 4 : HCT 116 (human colorectal carcinoma epithelial cell) whole cell 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: 40 kDa
    Observed band size: 33,42 kDa why is the actual band size different from the predicted?



    The molecular weight observed is consistent with what has been described in the literature (PMID: 29505568).

    The lower 33-kDa band is likely to be a truncated form (PMID: 29505568).

    Control cell line: HCT-116 (low-expression, PMID: 27016617).

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1: 5 seconds; Lane 2, 3 and 4: 37 seconds.

  • Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)
    Anti-LINE-1 ORF1p antibody [EPR22227-6] (ab245249)

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