Source: The AGE modified BSA was produced by reacting BSA with glucose under sterile conditions for 2 months followed by dialysis and purification according to the method described in Valentia et al. (2004). The AGE-BSA was biotinylated and then subjected to dialysis.

Formulation:

Frozen liquid in phosphate-buffered saline (PBS), 1 mg/mL

Endotoxin:

< 0.1 EU/μg AGE-BSA as determined by the LAL method

Preservative:

None

Sterility:

0.2 um membrane-filtered and packaged aseptically.

QC tests:

SDS-PAGE, MALDI-TOF, Fluorescence spectrometry, Absorbance at 320 nm and RAGE binding ELISA

Storage and stability:

Upon receiving, store the product at -20°C. After thawing, store the working aliquots at 2-8°C for up to 1 month. For extended storage, aliquot the solution and freeze at -70°C or -20°C. Avoid repeated freezing and thawing.

Activity:

KD 50-200 nM, as measured by its binding to the immobilized RAGE in an ELISA binding assay

Extent of AGE modification:

25-30 adducts per Biotin-AGE-BSA, as judged by free amine content analysis and MALDI-TOF analysis.

Extent of biotinylation:

6-7 biotins per AGE-BSA, as judged by HABA assay and MALDI-TOF analysis

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About Bioin-AGE-BSA:

Advanced glycation end products (AGEs) accumulate with age and at an accelerated rate in diabetes. AGEs bind cell-surface receptors including the receptor for AGE (RAGE). The fluorescence of Biotin-AGE-BSA was determined by fluorescence spectrometry with Ex./Em.=360/465 nm. The Biotin-AGE-BSA shows a 100-fold increase in fluorescence when compared to the control BSA. Browning of Biotin-AGE-BSA was determined by measuring the absorbance at 320 nm. The Bioin-AGE-BSA shows a 40 fold increase in absorbance when compared to the control BSA.

Reference:

Valencia et al. (2004) Anal. Biochem. 324:68-78