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).
Use at an assay dependent concentration.
Involved in cytoskeletal rearrangements required for phagocytosis of apoptotic cells and cell motility. Acts in assocation with DOCK1 and CRK. Was initially proposed to be required in complex with DOCK1 to activate Rac Rho small GTPases. May enhance the guanine nucleotide exchange factor (GEF) activity of DOCK1.
Widely expressed, with a higher expression in the spleen and placenta.
Contains 1 ELMO domain. Contains 1 PH domain.
Phosphorylated by HCK.
Cytoplasm. Cell membrane. Translocation to plasma membrane seems to be mediated by DOCK1 and CRK.
IHC image of ab2239 staining in human normal lymphoid formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab2239, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - ELMO1 antibody (ab2239)
Ab2239 staining (1µg/ml) of Jurkat lysate (RIPA buffer, 35µg total protein per lane).Primary incubated for 1 hour.Detected by western blot using chemiluminescence. Ab2239 staining (1µg/ml) of Jurkat lysate (RIPA buffer, 35µg total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence.
Immunoprecipitation - ELMO1 antibody (ab2239)This image is courtesy of an anonymous Abreview
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.
Immunocytochemistry/ Immunofluorescence - ELMO1 antibody (ab2239)Image from Sai Jiqing et. al. J. Biol. Chem., Sep 2008; 283: 26538 - 26547 (Fig 4A).
ab2239 staining ELMO1 in human HL60 cells by Immunocytochemistry/ Immunofluoresecence. Cells were fixed in 4% paraformaldehyde for 10 minutes, permeabilizedin 0.2% Triton X-100 in PBS for 5 minutes, blocked in 10% normaldonkey 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 coverslipswere incubated with fluorescence-conjugated secondary antibodiesfor 1 hour.
Makino Y et al. Tyr724 phosphorylation of ELMO1 by Src is involved in cell spreading and migration via Rac1 activation. Cell Commun Signal13:35 (2015).
Read more (PubMed: 26205662) »
Capala ME et al. ELMO1 is upregulated in AML CD34+ stem/progenitor cells, mediates chemotaxis and predicts poor prognosis in normal karyotype AML. PLoS One9:e111568 (2014).
Read more (PubMed: 25360637) »