Fibronectin HiLyte488™ (bovine plasma)

AT

1-2 Weeks

15

Fibronectin is purified from bovine plasma. Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 4-20% polyacrylamide gel. HiLyte Fluor 488 labeled fibronectin is >80% pure (Figure 1).
The protein is modified to contain covalently linked HiLyte Fluor 488 at random surface lysines (2). An activated ester of the fluorochrome is used to label the protein. Labeling stoichiometry is determined by spectroscopic measurement of protein and dye concentrations. Final labeling stoichiometry is 1-3 dyes per protein molecule (Figure 2). HiLyte Fluor 488 labeled fibronectin can be detected using a filter set of 350-450nm excitation and 500-550 nm emission.
Fibronectin runs as individual subunits on SDS-PAGE with an apparent molecular weight of 230 kDa. FNR02 is supplied as an orange lyophilized powder. Each vial of FNR02 contains 20 ug protein.

Figure 2: Absorption scan of HiLyte Fluor 488 labeled fibronectin in solution. FNR02 was diluted with Milli-Q water and its absorbance spectrum was scanned between 250 and 650 nm. HiLyte Fluor 488 labeling stoichiometry was calculated to be 1-3 dyes per fibronectin protein using the absorbancy maximum for at 527 nm and the Beer-Lambert law. Dye extinction coefficient when protein bound is 70, 000M-1cm-1