Western blot analysis of extracts from HL-60 cells, untreated or treated with retinoic acid for the indicated times (RA; 1 μM), using APR3 Antibody.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with an expression construct encoding tagged APR3 (+), using APR3 Antibody. The construct was kindly provided by Drs. Timothy Peterson and Erin O'Shea, Harvard University, Boston MA.
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
APR3 Antibody recognizes endogenous levels of total APR3 protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro29 of human APR3 protein (isoform a). Antibodies are purified by protein A and peptide affinity chromatography.
Apoptosis related protein 3 (APR3), also known as C2orf28, is a membrane protein identified in HL-60 cells treated with all-trans retinoic acid (ATRA) and was later found to be induced by ATRA in other sensitive cell lines (1,2). APR3 is also up-regulated by NFAT and NF-κB activities (3). Regulation of APR3 by ATRA suggests a role in cell differentiation, but the mechanism of action is still unclear. Overexpression of APR3 can inhibit proliferation by inducing G1/S cell cycle arrest and decreasing expression of cyclin D1 (2). APR3 has also been shown to interact with NELL-1 to regulate osteoblast differentiation (4).
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