Purified from low density lipoproteins (LDL) isolated by density gradient ultracentrifugation (1.03 - 1.05 g/mL density range) and by gel filtration chromatography in 10 mM sodium deoxycholate, 50 mM NaCl, 50 mM Na2CO3, pH 10.1.
Density gradient ultracentrifugation and gel filtration chromatography
>= 95% by SDS-PAGE
Lyophilized from 1 ml of 10 mM sodium deoxycholate, 50 mM NaCl, 5 0mM Na2CO3, pH 10
Store at -20°C or lower on dry atmosphere. After reconstitution, store at 2-8°C no longer than 4 weeks. Reconstitution: Dissolve lyophilized Apo B in 100 mL to 1 mL of distilled or deionized water. If dissolving in solvent we recommend using 10 mM sodium deoxycholate, 50 mM NaCl, 50 mM Na2CO3, pH 10 to keep the sodium deoxycholate at an acceptable level instead of water.
Universal Precautions. Safe laboratory practices should be used when handling this material.
The Apo B prepared by this procedure is aggregated. The aggregates are not retained by a 0.45 um filter, but migrate only a short distance into a 5% SDS-PAGE gel. In the isolation buffer, Apo B gives a stable solution that is slightly hazy.
This gene product is the main apolipoprotein of chylomicrons and low density lipoproteins. It occurs in plasma as two main isoforms, apoB-48 and apoB-100: the former is synthesized exclusively in the gut and the latter in the liver. The intestinal and the hepatic forms of apoB are encoded by a single gene from a single, very long mRNA. The two isoforms share a common N-terminal sequence. The shorter apoB-48 protein is produced after RNA editing of the apoB-100 transcript at residue 2180 (CAA->UAA), resulting in the creation of a stop codon, and early translation termination. Mutations in this gene or its regulatory region cause hypobetalipoproteinemia, normotriglyceridemic hypobetalipoproteinemia, and hypercholesterolemia due to ligand-defective apoB, diseases affecting plasma cholesterol and apoB levels. [provided by RefSeq