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Synthetic peptide conjugated to KLH derived from within residues 800 to the C-terminus of Human hnRNP U.
(Peptide available as ab21993.)
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.
|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|
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.
Image courtesy of Human Protein Atlas
ab20666 staining hnRNP U/p120 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/p120 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.
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