Antibodies, Proteins, Kits and Reagents for Life Science

314 937 ₸ Product size

100 µl 314 937 ₸

Order now and get it on Tuesday March 02, 2021

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

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.

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

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)

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)

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)

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)

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)

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 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)

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)

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