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Our Abpromise guarantee covers the use of ab2239 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 at an assay dependent concentration. PubMed: 18662984|
|IHC-P||Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|WB||Use a concentration of 0.1 - 0.3 µg/ml. Can be blocked with Human ELMO1 peptide (ab22878).
Note the smaller isoform of this protein has a consensus glycosylation site, which may explain the higher than expected band size (50 kDa versus 30 kDa).
|IP||Use at an assay dependent concentration.|
Primary incubation was 1 hour. Detected by chemiluminescence.
Lane 1-3: whole cell lysate (differentiated HL60 cells)
Lane 4: Protein molculare standard (100, 75 and 50kDa)
Lane 5: Immunoprecipitation with normal IgG (mock)
Lane 6 and 7: Immunoprecipitation with ab2239 antibody
Immunoprecipiated with ab2239 1.5 ug/sample (about 400 ug total protein). Immunoblotted with polyclonal rabbit anti-Dock2 or ab2239 antibody.
ab2239 staining ELMO1 in human HL60 cells by Immunocytochemistry/ Immunofluoresecence. Cells were fixed in 4% paraformaldehyde for 10 minutes, permeabilized in 0.2% Triton X-100 in PBS for 5 minutes, blocked in 10% normal donkey serum for 30 minutes and incubated with primary antibodies for 2 hours at room temperature. After washing three times with 0.1% Tween 20 in PBS, the coverslips were incubated with fluorescence-conjugated secondary antibodies for 1 hour.