Western blot analysis of extracts from various cell lines using Mis18-α (D3N2X) Rabbit mAb.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected (+) with full length human Mis18-α/Myc-DDK, using Mis18-α (D3N2X) Rabbit mAb (upper) or Myc-Tag (71D10) Rabbit mAb #2278 (lower).
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. 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
Mis18-α (D3N2X) Rabbit mAb recognizes endogenous levels of total Mis18-α protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg41 of human Mis18-α protein.
CENP-A, also known as the chromatin-associated protein CSE4 (capping-enzyme suppressor 4-p), is an essential histone H3 variant that replaces canonical histone H3 in centromeric heterochromatin. The inherited localization of the centromere is specified by CENP-A (1). CENP-A deposition to the correct chromosomal location in early G1 phase is regulated by the Mis18 complex, which consists of Mis18-alpha, Mis18-beta, Mis18BP1, RbAp48 and RbAp46 (2).
Mis18-alpha deficiency in mice results in inappropriate localization of CENP-A, as well as DNA methylation defects (3). Localization of the Mis18 complex to centromeres is regulated by the mitotic kinase Plk1 (polo-like kinase 1) (4).
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