Western blot analysis of extracts from OVCAR-4 and HCT 116 cells using Asparagine Synthetase 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
Asparagine Synthetase Antibody recognizes endogenous levels of total asparagine synthetase protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human asparagine synthetase protein. Antibodies are purified by protein A and peptide affinity chromatography.
Asparagine synthetase (ASNS) catalyzes the synthesis of asparagine from aspartate and glutamine. Research studies have shown that intracellular asparagine can suppress apoptosis in a large number of human tumors. In addition, ASNS expression levels have been associated with the progression of gliomas and neuroblastomas in patients (1). Furthermore, acute lymphocytic leukemia cells frequently depend upon serum asparagine for their viability, as they lack ASNS (2). Deprivation of asparagine by L-asparaginase has therefore been developed as a therapeutic treatment for acute lymphocytic leukemia (2-3). In subsets of gastric and hepatic cancers, ASNS promoter hypermethylation correlates with low ASNS expression, sensitizing these cancers to the asparaginase treatment (4).
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