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: IHC-Fr, WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Full length protein corresponding to Mouse LINE-1 ORF1p aa 1 to the C-terminus.
    Database link: P11260

  • Positive control
    • WB: F9 whole cell lysate and mouse testis lysate. IHC-P: Mouse testis tissue. ICC/IF: F9 cells. Flow Cyt: F9 cells. IP: F9 whole cell lysate.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

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
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR21844-108
  • Isotype
    IgG
  • Research areas

Applications

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
IHC-Fr 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

  • 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).

  • ab216324 staining LINE-1 ORF1p in Wt and Maelstrom (Mael) null mutant mouse testis by Immunohistochemistry (Frozen sections). Mael is a repressor of LINE-1 transposable elements, so the expression of LINE-1 ORF1p increases in this KO. Tissue was fixed with paraformaldehyde and blocked with 10% serum for 30 minutes at 25°C; Samples were incubated with primary antibody (1/100, 1/500 and 1/1000) for 12 hours at 4°C in 3% Donkey Serum in PBS + 0.05% Triton X. Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073) (1/1000) was used as the secondary antibody. 

    See Abreview

  • 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).

  • 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.

  • 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).

  • 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 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).

  • 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.

References

ab216324 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

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

Dr. Alex Bortvin

Verified customer

Submitted Feb 11 2019

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

Steven Cincotta

Verified customer

Submitted Jan 08 2019

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