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Recombinant full length protein corresponding to hnRNP U. RNPs eluted from an oligo (dt) cellulose column
Our Abpromise guarantee covers the use of ab10297 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration. Detects a band of approximately 97 kDa (predicted molecular weight: 120 kDa).|
|IP||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
U-2 OS (human bone osteosarcoma epithelial cell line) cells stained for hnRNP U (green) using ab10297 (1/100 dilution) in ICC/IF. The cells were paraformaldehyde fixed and blocked with BSA prior to incubation with the antibody for 30 minutes. A FITC conjugated anti-mouse antibody was used as the secondary. The cells were counterstained with DAPI (left hand panel).
This image is courtesy of an Abreview submitted by Dr Stoil Dmitrov
ab10297 (4 µg/ml)) staining hnRNPU in human cerebellum using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining of Purkinje cells neurons.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with a protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab10297 (red line). The cells were fixed with 100% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab10297, 1 µg/1x106 cells) for 30 minutes at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 minutes)/permeabilized in 0.1% PBS-Tween used under the same conditions.
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