Western blot analysis of extracts from Neuro2A and PC-12 cells, using MBTPS2 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 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 June 2020
Protocol Id: 10
MBTPS2 Antibody detects endogenous levels of total MBTPS2 protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of mouse MBTPS2. Antibodies are purified by protein A and peptide affinity chromatography.
Membrane-bound transcription factor protease site 2 (MBTPS2), also known as site-2 protease (S2P), is a zinc metalloprotease in the Golgi membrane (1,2,3). It regulates cholesterol metabolism (1,2) and unfolded protein response (UPR) (3,4). When cells are deprived of cholesterol, sterol regulatory element-binding proteins (SREBPs) move from the endoplasmic reticulum (ER) to the Golgi apparatus and are cleaved by site-1 protease (S1P) (5,6) and site-2 protease (1,6) sequentially to release the active amino-terminal domains. These amino-terminal domains of SREBPs then translocate into the nucleus to induce expression of genes for cholesterol biosynthesis. During UPR, activating transcription factor 6 (ATF6) transports from ER to Golgi apparatus and is cleaved by S1P and S2P to release a cytosolic fragment. This cytosolic fragment relocates to the nucleus and activates the UPR gene expression (7).
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