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RecombinantRabMAb

Recombinant Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)

Reviews (2) Submit a question References (7)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
  • Western blot - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
  • Western blot - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
  • Immunocytochemistry/ Immunofluorescence - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
  • Flow Cytometry (Intracellular) - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
  • Immunoprecipitation - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
  • Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21844-108] to LINE-1 ORF1p
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
  • Reacts with: Mouse

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free

Overview

  • Product name

    Anti-LINE-1 ORF1p antibody [EPR21844-108]
    See all LINE-1 ORF1p primary antibodies

  • Description

    Rabbit monoclonal [EPR21844-108] to LINE-1 ORF1p

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Mouse

  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: F9 whole cell lysate and mouse testis lysate. IHC-P: Mouse testis tissue. ICC/IF: F9 cells. Flow Cyt (intra): F9 cells. IP: F9 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

    EPR21844-108

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab216324 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
Flow Cyt (Intra)
1/500.
WB
1/1000. Detects a band of approximately 41 kDa (predicted molecular weight: 41 kDa).
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/100.
IP
1/30.
Notes
Flow Cyt (Intra)
1/500.
WB
1/1000. Detects a band of approximately 41 kDa (predicted molecular weight: 41 kDa).
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/100.
IP
1/30.

Target

  • Relevance

    Domain: The coiled coil domain mediates homotrimerization. The RRM and the CTD domain are both required for proper RNA-binding activity. Function: Nucleic acid-binding protein which is essential for retrotransposition of LINE-1 elements in the genome. Functions as a nucleic acid chaperone binding its own transcript and therefore preferentially mobilizing the transcript from which they are encoded. Long interspersed element-1/LINE-1/L1 retrotransposons are present in more than 500'000 full (6 kb) or truncated copies in the human genome. Most of them are inactive but 80 to 100 of those elements could be transcribed, translated and active in any individual. An active LINE-1 encodes for 2 proteins translated from a single RNA containing 2 non-overlapping ORFs, ORF1 and ORF2. ORF1p is described in this entry as a representative of all ORF1p potentially expressed by active elements. ORF2p is described in the related entry AC O00370. PTM: Polyubiquitinated, probably by UBR2, which induces its degradation. Similarity: Belongs to the transposase 22 family.

  • Database links

    • Alternative names

      • L1ORF1p antibody
      • L1RE1 antibody
      • LINE retrotransposable element 1 antibody
      • LINE-1 retrotransposable element ORF1 protein antibody
      • LINE1 retrotransposable element 1 antibody
      • LRE1 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)

      Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling LINE-1 ORF1p with ab216324 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on spermatogonia of mouse testis (PMID: 24607009) is observed. 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).

    • Western blot - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
      Western blot - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
      All lanes : Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324) at 1/1000 dilution

      Lane 1 : F9 (mouse embryonal carcinoma epithelial cell), whole cell lysate
      Lane 2 : NIH/3T3 (mouse embryonic fibroblast), 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: 41 kDa
      Observed band size: 41 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      Negative control: NIH/3T3 (PMID:24324740).

    • Western blot - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
      Western blot - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
      Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324) at 1/1000 dilution + mouse testis lysate at 20 µg

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

      Predicted band size: 41 kDa
      Observed band size: 41 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)

      Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling LINE-1 ORF1p with ab216324 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Negative control: No staining on mouse liver (PMID: 24607009) is observed. Counterstained with hematoxylin.
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunocytochemistry/ Immunofluorescence - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
      Immunocytochemistry/ Immunofluorescence - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labeling LINE-1 ORF1p with ab216324 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 staining in F9 cell line.

      Negative control: NIH/3T3 (PMID: 24324740).

      The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).

    • Flow Cytometry (Intracellular) - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
      Flow Cytometry (Intracellular) - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast, Left) / F9 (mouse embryonal carcinoma epithelial cell, Right) cell lines labeling LINE-1 ORF1p with ab216324 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. 

      Negative control: NIH/3T3 (PMID:24324740). 

       

       

    • Immunoprecipitation - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
      Immunoprecipitation - Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)

      LINE-1 ORF1p was immunoprecipitated from 0.35 mg F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate with ab216324 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216324 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution

      Lane 1: F9 whole cell lysate 10 µg (Input).
      Lane 2: ab216324 IP in F9 whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216324 in F9 whole cell lysate.

      Blocking/Dilution buffer: 5% NFDM/TBST.
      Exposure time: 30 seconds.

    • Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)
      Anti-LINE-1 ORF1p antibody [EPR21844-108] (ab216324)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (7)

    Publishing research using ab216324? Please let us know so that we can cite the reference in this datasheet.

    ab216324 has been referenced in 7 publications.

    • Garland W  et al. Chromatin modifier HUSH co-operates with RNA decay factor NEXT to restrict transposable element expression. Mol Cell 82:1691-1707.e8 (2022). WB ; Mouse . PubMed: 35349793
    • Zhang Y  et al. Single-cell epigenome analysis reveals age-associated decay of heterochromatin domains in excitatory neurons in the mouse brain. Cell Res N/A:N/A (2022). Mouse . PubMed: 36207411
    • Wang F  et al. Inhibition of LINE-1 retrotransposition represses telomere reprogramming during mouse 2-cell embryo development. J Assist Reprod Genet N/A:N/A (2021). Mouse . PubMed: 34618297
    • Müller I  et al. MPP8 is essential for sustaining self-renewal of ground-state pluripotent stem cells. Nat Commun 12:3034 (2021). WB ; Mouse . PubMed: 34031396
    • Chini CCS  et al. CD38 ecto-enzyme in immune cells is induced during aging and regulates NAD+ and NMN levels. Nat Metab 2:1284-1304 (2020). IHC-Fr ; Mouse . PubMed: 33199925
    • Shirane K  et al. NSD1-deposited H3K36me2 directs de novo methylation in the mouse male germline and counteracts Polycomb-associated silencing. Nat Genet N/A:N/A (2020). IHC-Fr ; Mouse . PubMed: 32929285
    • Guerrero A  et al. Cardiac glycosides are broad-spectrum senolytics. Nat Metab 1:1074-1088 (2019). PubMed: 31799499

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-LINE-1 ORF1p antibody [EPR21844-108]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Wt and Maelstrom null mutant testis)
    Permeabilization
    Yes - 0.1% Triton X-100 in PBS
    Specification
    Wt and Maelstrom null mutant testis
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Alex Bortvin

    Verified customer

    Submitted Feb 11 2019

    Immunohistochemistry (Frozen sections) abreview for Anti-LINE-1 ORF1p antibody [EPR21844-108]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Testis, embryonic day 17.5)
    Permeabilization
    Yes - PBS + 0.5% Triton X-100
    Specification
    Testis, embryonic day 17.5
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Steven Cincotta

    Verified customer

    Submitted Jan 08 2019

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