Overview

  • Product name
  • Description
    Rabbit polyclonal to hnRNP U
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-P, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 800 to the C-terminus of Human hnRNP U.

    (Peptide available as ab21993.)

  • Positive control
    • This antibody gave a positive signal in HeLa cells, and HepG2 cells, and HeLa whole cell lysate.

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab20666 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
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 117 kDa (predicted molecular weight: 90 kDa).
IHC-P 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
ChIP Use at an assay dependent concentration. PubMed: 21235343

Target

  • Function
    Component of the CRD-mediated complex that promotes MYC mRNA stabilization. Binds to pre-mRNA. Has high affinity for scaffold-attached region (SAR) DNA. Binds to double- and single-stranded DNA and RNA.
  • Sequence similarities
    Contains 1 B30.2/SPRY domain.
    Contains 1 SAP domain.
  • Post-translational
    modifications
    Extensively phosphorylated.
    Arg-733 and Arg-739 are dimethylated, probably to asymmetric dimethylarginine.
  • Cellular localization
    Nucleus. Cytoplasm. Cell surface. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Component of ribonucleosomes. Also found associated with the cell surface.
  • Information by UniProt
  • Database links
  • Alternative names
    • Heterogeneous nuclear ribonucleoprotein U antibody
    • hnRNP U antibody
    • hnRNP U protein antibody
    • HNRNPU antibody
    • hnRNPU protein antibody
    • HNRPU antibody
    • HNRPU_HUMAN antibody
    • p120 antibody
    • p120 nuclear protein antibody
    • pp120 antibody
    • SAF A antibody
    • SAF-A antibody
    • SAFA antibody
    • Scaffold attachment factor A antibody
    • U21.1 antibody
    see all

Images

  • ICC/IF image of ab20666 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab20666, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

    Panel A shows localisation of ab20666 to the nuclei, Panel B has the Alexa Fluor® 488 channel removed for comparison.

  • Anti-hnRNP U antibody (ab20666) at 1 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 90 kDa
    Observed band size: 117 kDa (why is the actual band size different from the predicted?)

  • Image courtesy of Human Protein Atlas

    ab20666 staining hnRNP U in paraffin embedded human skin tissue.  The samples were incubated with ab20666 (1/250 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab20666 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

    Further results for this antibody can be found at ww

  • ab20666 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab20666 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Immunofluorescence analysis of murine embryonic stem cells, staining hnRNP U with ab20666.

    Cells were fixed with paraformaldehyde, permeabilized with 0.25% Triton X-100 and blocked in 0.1% Triton X-100/10% FCS for 20 min. Samples were incubated with primary antibody (1/500) for 2 hours before incubating with an AlexaFluor®488-conjugated goat anti-rabbit IgG (1/500) for 1 hour.

References

This product has been referenced in:
  • Lu Y  et al. The NF-?B-Responsive Long Noncoding RNA FIRRE Regulates Posttranscriptional Regulation of Inflammatory Gene Expression through Interacting with hnRNPU. J Immunol 199:3571-3582 (2017). Read more (PubMed: 28993514) »
  • Zheng Q  et al. Nuclear distribution of eIF3g and its interacting nuclear proteins in breast cancer cells. Mol Med Rep 13:2973-80 (2016). WB . Read more (PubMed: 26935993) »

See all 11 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Heat-inactivated normal donkey serum in 0.05% PBS-T as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
None
Sample
Mouse Tissue sections (Mouse, whole brain sections, striatum)
Specification
Mouse, whole brain sections, striatum
Permeabilization
Yes - Tween-20
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 07 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (4-12% Bis-Tris Gel, MES running buffer)
Sample
Mouse Tissue lysate - whole (Mouse whole brain tissue lysate, striatum)
Specification
Mouse whole brain tissue lysate, striatum
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Sep 20 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Huh7 cell (Hepatocarcinoma))
Loading amount
100 µg
Specification
Huh7 cell (Hepatocarcinoma)
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Sep 26 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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