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Amplite® Fluorimetric Fluorescamine Protein Quantitation Kit *Blue Fluorescence*

BSA dose response was measured on a solid black 96-well plate with Amplite® Fluoremetric Fluorescamine Protein Quantitation Assay Kit.
BSA dose response was measured on a solid black 96-well plate with Amplite® Fluoremetric Fluorescamine Protein Quantitation Assay Kit.
BSA dose response was measured on a solid black 96-well plate with Amplite® Fluoremetric Fluorescamine Protein Quantitation Assay Kit.
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Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22


Fluorescamine is intrinsically non-fluorescent but reacts rapidly with primary aliphatic amines, including those in peptides and proteins, to yield a blue-green-fluorescent derivative. The Amplite® fluorescamine protein assay kit provides a simple method for quantifying protein concentration in solutions. This Amplite® fluorescamine protein assay kit can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation with no separation steps required. The assay can be completed within 30 minutes. With the Amplite® fluorescamine protein assay kit, as little as 3 ug/mL of BSA can be detected.


Fluorescence microplate reader

Recommended plateSolid black


Example protocol


Protocol summary

  1. Prepare  fluorescamine working solution (25 µL)
  2. Add BSA standards or test samples (75 µL)
  3. Incubate at room temperature for 5 - 30 minutes
  4. Read fluorescence intensity at Ex/Em = 380/470 nm

Important notes
Thaw all the kit components at room temperature before starting the experiment.


BSA standard

For convenience, use the Serial Dilution Planner: https://www.aatbio.com/tools/serial-dilution/11100

Dilute the appropriate amount of BSA Standard 1 mg/mL (Component C) into PBS by performing 1:2 serial dilutions to get serial dilutions of BSA standard (BS7 - BS1).


Add the whole content of DMSO (Component B) into the bottle of Fluorescamine (Component A), and mix well. Note: 2.5 mL of fluorescamine working solution is enough for 1 plate.


Table 1. Layout of BSA standards and test samples in a solid black 96-well microplate. BS= BSA Standards (BS1 - BS7, 1.563 to 100 µg/mL), BL=Blank Control, TS=Test Samples.


Table 2. Reagent composition for each well.

BS1 - BS775 uLSerial Dilution (1.563 to 100 ug/mL)
TS75 uLTest Sample
  1. Prepare BSA standards (BS), blank controls (BL), and test samples (TS) according to the layout provided in Tables 1 and 2. For a 384-well plate, use 30 µL of reagent per well instead of 75 µL.

  2. Add 25 µL of  fluorescamine working solution to each well of BSA standard, blank control, and test samples to make the total assay volume of 100 µL/well. For a 384-well plate, add 10 µL of fluorescamine working solution into each well instead, for a total volume of 40 µL/well.

  3. Incubate the reaction at room temperature for 5 to 30 minutes, protected from light.

  4. Monitor the fluorescence increase with a fluorescence plate reader at Ex/Em = 380/470 nm.



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Covalent vaccination with Trypanosoma cruzi Tc24 induces catalytic antibody production
Authors: Gunter, Sarah M and Versteeg, Leroy and Jones, Kathryn M and Brian, Keegan P and Strych, Ulrich and Bottazzi, Maria Elena and Hotez, Peter J and Brown, Eric L
Journal: Parasite immunology (2018): e12585


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Journal: J Fluoresc (2005): 679
Determination of 4-amino-m-cresol and 5-amino-o-cresol by high performance liquid chromatography and fluorescence derivatization using fluorescamine
Authors: Eggenreich K, Zach E, Beck H, Wintersteiger R.
Journal: J Biochem Biophys Methods (2004): 35
Chiral separation of fluorescamine-labeled amino acids using microfabricated capillary electrophoresis devices for extraterrestrial exploration
Authors: Skelley AM, Mathies RA.
Journal: J Chromatogr A (2003): 191
LC determination of aminoglutethimide enantiomers as dansyl and fluorescamine derivatives in tablet formulations
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Journal: J Pharm Biomed Anal (2002): 487
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Journal: J Pharm Biomed Anal (2002): 253
A comparison of fluorescamine and naphthalene-2,3-dicarboxaldehyde fluorogenic reagents for microplate-based detection of amino acids
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Journal: Anal Biochem (2001): 128
Detection, quantitation, and identification of residual aminopenicillins by high-performance liquid chromatography after fluorescamine derivation
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Journal: J Food Prot (2000): 1421
Determination of taurine in plasma by capillary zone electrophoresis following derivatisation with fluorescamine
Authors: Kelly MT, Fabre H, Perrett D.
Journal: Electrophoresis (2000): 699
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