Western blot analysis of extracts from various cell lines using ACF1 Antibody.
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
ACF1 Antibody recognizes endogenous levels of total ACF1 protein (isoforms 1 and 2).
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Met864 of human ACF1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
The mammalian imitation SWI (ISWI) complexes are characterized by two ATPase subunits: Snf2h and Snf2l (1). Snf2h interacts with ATP-utilizing chromatin assembly and remodeling factor 1 (ACF1) to comprise the ACF chromatin-remodeling complex (1). ACF1 (BAZ1A) has distinct roles in development (2), regulation of chromatin structure (3), and DNA damage response (4,5). Different developmental stages dictate the expression of ACF1 in Drosophila, and alterations in ACF1 expression during Drosophila development leads to deviation from normal chromatin organization (2). ACF1 functions in heterochromatin formation during development and is involved in the initial establishment of diversified chromatin structures. In vivo studies demonstrate that heterochromatin protein 1 (HP1) binding to methylated lysine 9 of histone H3 is enhanced by the interaction of ACF1 with chromatin (6). Chromatin-remodeling factors are required during DNA damage in order to allow signaling molecules and damaging enzymes to access the site (4). Depletion of hACF1 increases apoptosis and vulnerability to radiation and compromises G2/M arrest activated in response to X-ray and UV exposure (4). Depletion of ACF1 also sensitizes cells to DNA double-stranded breaks (DSBs) and impairs DNA repair (5). Specifically, accumulation of Ku at DSBs sites may depend on the presence of ACF1 (5).
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