Western blot analysis of extracts from various cell lines using PRDX6 (D9J9H) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (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 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
PRDX6 (D9J9H) Rabbit mAb recognizes endogenous levels of total PRDX6 protein.Species Reactivity:
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human PRDX6 protein.
Peroxiredoxin 6 (PRDX6) belongs to an antioxidant enzyme family of non-seleno peroxidases. PRDX6 is a unique member of the PRDX family, exhibiting both glutathione peroxidase and phospholipase A2 activities (1,2). PRDX6 regulates phospholipid turnover in addition to protecting cells against oxidative injury. PRDX6 is expressed in all major organs, with a particularly high level in lung, where it regulates lung surfactant phospholipid synthesis and turnover (3-5). Research studies have shown that PRDX6 is aberrantly expressed in various cancers, and can promote cancer cell metastasis and invasion (6,7). Elevated expression of PRDX6 and related PRDX family members has also been shown to contribute to drug resistance in cancer cells (8,9). PRDX6 is also expressed in neutrophils, where it has been shown to activate NADPH oxidase (NOX2) (10-12).
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