PROTEIN ASSAY OF ONE SAMPLE

University Of Georgia, Athens GA

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Abstract

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Ammonium Sulfate Precipitation The Enoxaparin sodium solution (150 mg/mL) was divided into three 500-mL portions. Two of the portions were spiked with BSA (1 and 5 uL, respectively of a 2 mg/mL solution). Two of three 500-uL DI water blanks were spiked in the same way. Each of the six solutions was treated with 280 mg ammonium sulfate (=80 % saturation), vortexed until homogeneous, and incubated at 25 [unreadable]C for 1 h. The samples were centrifuged for 15 min at 16,000 g, and the supernatant was removed. Bradford Dye-Binding Protein Assay Each pellet was dissolved in 800 uL DI water, and 200 uL Bio-Rad Bradford reagent concentrate was added. After vortexing, the samples were transferred to 10-mm microcuvettes, and absorbance at 595 nm was measured.