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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Ikaros.
(Peptide available as ab35526.)
Our Abpromise guarantee covers the use of ab26083 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 63 kDa (predicted molecular weight: 56 kDa).|
|ICC/IF||Use a concentration of 5 µg/ml.|
The lower band size observed in spleen corresponds to Isoform 6 of the protein.
It is believed that the higher bands observed in Hela and Jurkat lysates may be due to aberrant expression in cancer cell lines.
Sun L et al. Expression of aberrantly spliced oncogenic ikaros isoforms in childhood acute lymphoblastic leukemia. J Clin Oncol 17:3753-66 (1999). PMID:10577847.
ab26083 stained in MOLT4 cells. Cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab26083 at 5 µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.
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