Western blot analysis of extracts from various cell lines using Regucalcin/SMP30 Antibody (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Expression levels of regucalcin/SMP30 among cell lines are consistent with expectations based on publicly available bioinformatic databases.
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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 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
Regucalcin/SMP30 Antibody recognizes endogenous levels of total regucalcin/SMP30 protein.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human regucalcin/SMP30 protein. Antibodies are purified by peptide affinity chromatography.
Regucalcin is a highly conserved, multifunctional protein encoded by the RGN gene on the X chromosome, generally recognized as a calcium (Ca2+) binding protein involved in the maintenance of intracellular Ca2+ homeostasis (1). The protein was independently identified and termed Senescence Marker Protein-30 (SMP30), following the observation in a rat aging model that expression of the protein markedly declined with age in a hormone-independent manner (2). Regucalcin/SMP30 has since been implicated in the regulatory control of many fundamental biological processes (e.g., cell growth, apoptosis, cellular metabolism), largely through its role in regulating Ca2+ homeostasis and Ca2+-dependent signal transduction (3-5). Research studies also suggest that regucalcin/SMP30 may function as a tumor suppressor in some contexts (3,6), while serum levels of regulcalcin/SMP30 have also been proposed as potential biomarkers of liver and kidney dysfunction (7).
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