Western blot analysis of extracts from various cell lines using ZIP8/SLC39A8 (E2L8E) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
|MW (kDa)||65, 130|
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 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
ZIP8/SLC39A8 (E2L8E) Rabbit mAb recognizes endogenous levels of total ZIP8/SLC39A8 protein.Species Reactivity:
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human ZIP8/SLC39A8 protein.
Zinc transporter zinc-and iron-related protein 8 (ZIP8) is encoded by the SLC39A8 gene and is a member of the solute carrier gene (SLC) superfamily (1). As a membrane protein, ZIP8 functions to transport manganese, cadmium, and zinc across the plasma membrane (2). Metal ions are essential elements that are required cofactors for proteins, including enzymes and transcription factors. Maintenance of proper metal ion homeostasis is required for normal mammalian tissue development and function. Mice lacking SLC39A8 reveal a critical role of ZIP8 in organogenesis in multiple tissues, including the brain and kidney, during development (3,4). Mutations in SLC39A8 are linked to autosomal-recessive forms of intellectual disability, suggesting that altered ZIP8-dependent metal ion regulation may contribute to human neuronal developmental diseases (5).
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