The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 117 kDa (predicted molecular weight: 90 kDa).
1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Use at an assay dependent concentration. PubMed: 21235343
FunctionComponent 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 similaritiesContains 1 B30.2/SPRY domain. Contains 1 SAP domain.
Post-translational modificationsExtensively phosphorylated. Arg-733 and Arg-739 are dimethylated, probably to asymmetric dimethylarginine.
Cellular localizationNucleus. Cytoplasm. Cell surface. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Component of ribonucleosomes. Also found associated with the cell surface.
Heterogeneous nuclear ribonucleoprotein U antibody
hnRNP U antibody
hnRNP U protein antibody
hnRNPU protein antibody
p120 nuclear protein antibody
SAF A antibody
Scaffold attachment factor A antibody
Anti-hnRNP U antibody - ChIP Grade images
Immunocytochemistry/ Immunofluorescence - hnRNP U antibody (ab20666)
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.
Western blot - hnRNP U antibody (ab20666)
Anti-hnRNP U antibody - ChIP Grade (ab20666) at 1 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 20 µg
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.
Immunocytochemistry/ Immunofluorescence - Anti-hnRNP U antibody (ab20666)
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.
Immunocytochemistry/ Immunofluorescence - Anti-hnRNP U antibody - ChIP Grade (ab20666)Image from Vizlin-Hodzic D et al., PLoS One. 2011;6(12):e28049. Epub 2011 Dec 6. Fig 1.; doi:10.1371/journal.pone.0028049; December 6, 2011, PLoS ONE 6(12): e28049.
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 for Anti-hnRNP U antibody - ChIP Grade (ab20666)
This product has been referenced in:
Shao R et al. Coordinate regulation of heterogeneous nuclear ribonucleoprotein dynamics by steroid hormones in the human fallopian tube and endometrium in vivo and in vitro. Am J Physiol Endocrinol Metab302:E1269-82 (2012).
Read more (PubMed: 22436695) »
Hope NR & Murray GI The expression profile of RNA-binding proteins in primary and metastatic colorectal cancer: relationship of heterogeneous nuclear ribonucleoproteins with prognosis. Hum Pathol42:393-402 (2011).
Read more (PubMed: 21194727) »