Western blot analysis of extracts from various cell lines using PEN2 Antibody.
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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
PEN2 Antibody detects endogenous levels of total PEN2 protein.Species Reactivity:
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys11 of human PEN2 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Presenilin Enhancer 2 (PEN2) is a small integral membrane glycoprotein that contains two recognized transmembrane domains. Both the N- and C-terminal domains are oriented into the lumen of the endoplasmic reticulum (1). PEN2, along with Presenilin 1, Presenilin 2, Nicastrin, and APH-1 form the protein complex γ-secretase (2). The proteinase BACE catalyses the initial step in APP processing by cleaving and releasing soluble APPβ (3). The remaining membrane bound APP is then cleaved by the γ-secretase complex, causing the release of amyloid β-peptide, the main constituent of amyloid plaques. These plaques are a hallmark of Alzheimer’s disease pathology (2). In addition to APP, the γ-secretase complex cleaves several other proteins and necessary presenilin-dependent signaling cascades, including the Notch pathway (4). It was found that PEN2 is an important part of the γ-secretase complex, and knocking it down results in reduced amounts of the complex, resulting in a loss of γ-secretase activity (5).
Explore pathways + proteins related to this product.
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