Recombinant
RabMAb

Recombinant Anti-PCBP2/hnRNP E2 antibody [EPR14859(2)] (ab200835)

Overview

  • Product name

    Anti-PCBP2/hnRNP E2 antibody [EPR14859(2)]
    See all PCBP2/hnRNP E2 primary antibodies
  • Description

    Rabbit monoclonal [EPR14859(2)] to PCBP2/hnRNP E2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, WB, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human PCBP2/hnRNP E2 aa 250-350. The exact sequence is proprietary.
    Database link: Q15366

  • Positive control

    • WB: HeLa, Jurkat, K562, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain, Mouse heart, Rat heart and Rat spleen lysates. ICC/IF: HeLa cells. Flow Cyt: Jurkat cells. IP: Jurkat 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

Applications

Our Abpromise guarantee covers the use of ab200835 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
IP 1/80.
WB 1/1000. Detects a band of approximately 35, 39 kDa (predicted molecular weight: 34-39 kDa).
ICC/IF 1/500.
Flow Cyt 1/250.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

Target

  • Function

    Single-stranded nucleic acid binding protein that binds preferentially to oligo dC. Major cellular poly(rC)-binding protein. Binds also poly(rU). Negatively regulates cellular antiviral responses mediated by MAVS signaling. It acts as an adapter between MAVS and the E3 ubiquitin ligase ITCH, therefore triggering MAVS ubiquitinationa and degradation.
  • Tissue specificity

    Detected in all tissues examined.
  • Sequence similarities

    Contains 3 KH domains.
  • Post-translational
    modifications

    Phosphorylated. The non-phosphorylated form(s) exhibited the strongest poly(rC)-binding activity.
  • Cellular localization

    Nucleus. Cytoplasm. Loosely bound in the nucleus. May shuttle between the nucleus and the cytoplasm.
  • Information by UniProt
  • Database links

  • Alternative names

    • Alpha CP2 antibody
    • Alpha-CP2 antibody
    • alphaCP-2 antibody
    • Cbp antibody
    • CTBP antibody
    • Heterogeneous nuclear ribonucleoprotein E2 antibody
    • Heterogenous nuclear ribonucleoprotein E2 antibody
    • hnRNP E2 antibody
    • hnRNP-E2 antibody
    • HNRNPE2 antibody
    • Hnrnpx antibody
    • HNRPE2 antibody
    • Hnrpx antibody
    • MGC110998 antibody
    • PCBP2 antibody
    • PCBP2_HUMAN antibody
    • poly(rC) binding protein 2 antibody
    • Poly(rC)-binding protein 2 antibody
    • Putative heterogeneous nuclear ribonucleoprotein X antibody
    • rCbinding protein 2 antibody
    see all

Images

  • All lanes : Anti-PCBP2/hnRNP E2 antibody [EPR14859(2)] (ab200835) at 1/10000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
    Lane 3 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 34-39 kDa
    Observed band size: 35,39 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Based on sequence analysis, ab200835 recognizes isoforms 1-6 which have corresponding Mw between 34-39kDa.

    Blocking/Dilution Buffer: 5% NFDM/TBST.

  • All lanes : Anti-PCBP2/hnRNP E2 antibody [EPR14859(2)] (ab200835) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Rat heart lysate
    Lane 4 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 34-39 kDa
    Observed band size: 35,39 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution Buffer: 5% NFDM/TBST.

  • All lanes : Anti-PCBP2/hnRNP E2 antibody [EPR14859(2)] (ab200835) at 1/1000 dilution

    Lane 1 : Rat heart lysate
    Lane 2 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 3 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 5 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 34-39 kDa
    Observed band size: 35,39 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking/Dilution Buffer: 5% NFDM/TBST.

  • All lanes : Anti-PCBP2/hnRNP E2 antibody [EPR14859(2)] (ab200835) at 1/10000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with PCBP2/hnRNP E2 peptide
    Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with PCBP1 peptide

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 34-39 kDa
    Observed band size: 35,39 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution Buffer: 5% NFDM/TBST.

    Based on sequence analysis, ab200835 (PCBP2/hnRNP E2) shares 80% homology with family member PCBP1. The levels of XR were tested in the accompanying blocking experiment.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PCBP2/hnRNP E2 with ab200835 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear and cytoplasmic staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows;
    -ve control 1: ab200835 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling PCBP2/hnRNP E2 with ab200835 at 1/250 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

  • PCBP2/hnRNP E2 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate with ab200835 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab200835 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: Jurkat whole cell lysate 10 µg (Input). Lane 2: ab200835 IP in Jurkat whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200835 in Jurkat whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 5 seconds

References

ab200835 has not yet been referenced specifically in any publications.

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