Western blot analysis of extracts from various cell lines using HYOU1 Antibody.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human HYOU1 (hHYOU1-Myc/DDK; +), using HYOU1 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 nonfat dry milk, 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.
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: 263
HYOU1 Antibody recognizes endogenous levels of total HYOU1 protein. This antibody reacts weakly with murine and rat HYOU1.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human HYOU1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Hypoxia up-regulated protein 1 (HYOU1, ORP150) is an endoplasmic reticulum (ER)-resident stress protein whose expression is upregulated in response to a variety of environmental conditions that promote ER stress (1,2). The amino-terminal region of HYOU1 displays extensive similarity to the ATPase domain of HSP70 family proteins, suggesting that HYOU1 contributes to maintaining proper protein folding and secretion under conditions that promote ER stress (2). Research studies have demonstrated that HYOU1 is cytoprotective and is involved in the inhibition of hypoxia-induced cell death (3).
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