Western blot analysis of extracts from various cell types using MYH Antibody.
Western blot analysis of extracts from COS-7 cells, mock transfected (-) or MYH-transfected (+), using MYH 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 datasheet or 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 November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised October 2016
Protocol Id: 10
MYH Antibody detects endogenous levels of total MYH protein. Based on the amino acid sequence, the antibody is expected to react with all isoforms of human MYH protein.Species Reactivity:
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to carboxy terminal residues of human MYH protein. Antibodies are purified using protein A and peptide affinity chromatography.
Base excision repair (BER) proteins catalyze the removal of incorrect or damaged bases, including oxidized bases, from DNA. N-glycosylases specific to a given lesion remove the incorrect base as the first step in BER. MYH is the mammalian ortholog of E. coli MutY, a DNA glycosylase that catalyzes the removal of 8-oxoG:A mismatches (1). Several MYH isoforms have been detected in human cells localizing to either the nucleus or the mitochondria (2). MYH interacts with DNA repair proteins and localizes to DNA damage foci after oxidative damage (3). Research studies have shown that mutations in the corresponding MYH gene are associated with human gastric (4) and colorectal (5-7) cancers.
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