Western blot analysis of extracts from various cell lines using MAOA Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). THP-1 cells contain a homozygous null mutation at the MAOA locus, confirming the specificity of the antibody for MAOA.
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
MAOA Antibody recognizes endogenous levels of total MAOA protein.Species Reactivity:
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro243 of human MAOA protein. Antibodies are purified by peptide affinity chromatography.
Monoamine oxidase A (MAOA), a mitochondrial enzyme, catalyzes the degradation of monoamines. Studies show that increased expression of MAOA promotes prostate cancer metastasis by activating the Shh-IL6-RANKL signaling network (1). In addition, MAOA expression in adipose tissue macrophages is upregulated in aging, leading to the reduction of noradrenaline availability in adipose tissue. Decreased levels of noradrenaline contribute to the aging-related decline of catecholamine-induced lipolysis in adipocytes (2). Furthermore, sympathetic neuron–associated macrophages import and degrade norepinephrine via the transporter SLC6A2 and MAOA, respectively, and thus contribute to obesity (3).
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