Nanodrop Spectrophotometer: Labeled Protein analysis (FITC-BSA)

Explains how to calculate ratio of FITC to BSA when analyzing a fluorescently-labeled protein

[Bioskill]

What if our protein (which is a synthesized 20 aa peptide) does not have W or Y residue to absorb at 280? Is this method still applicable?
In this case, should we consider the concentration of FITC peptide equal to the concentration of the protein? (as I said, the peptide is synthesized by the company and each peptide has only one FITC tag).

[kparunscientist]

Hi, we use 2000c Nanodrop to estimate antibody-PE conjugate. We use UV vis method in ND2000c. We see different Abs values at 280nm and 565nm for 1.5ul, 2ul and 3ul. We have an excel which calculate the conc. Of Ab and dye through these Abs values. We see a lot of variations between replicates and between analysts. Suggest if we are doing it wrong way. 1. Which method to use for Ab-dye sample. 2. Why there is a difference in Abs values with 1.5ul, 2ul and 3ul. 3. Why Abs values are not repetitive from a single sample. Thanks

[sruthireddy]

could you please comment on the procedure on how to conjugate fitc with BSA