Amplite® Universal Fluorimetric Protease Activity Assay Kit *Green Fluorescence*

Amplite® Universal Fluorimetric Protease Activity Assay Kit

Green Fluorescence

Monitoring of various protease activities has become a routine task for many biological laboratories. Our Amplite® Universal Fluorimetric Protease Activity Assay Kits are an ideal choice for performing routine assays necessary during the isolation of proteases, or for identifying the presence of contaminating proteases in protein samples. The kits use fluorescent casein conjugates that are proven to be a generic substrate for a broad spectrum of proteases. In the intact substrate, casein is heavily labeled with a fluorescent dye, resulting in significant fluorescence quenching. Protease-catalyzed hydrolysis relieves its quenching effect, yielding brightly fluorescent dye-labeled short peptides. The increase in fluorescence intensity is directly proportional to protease activity. The kits provide all the essential components with an optimized "mix & read" protocol that can be easily automated to HTS instruments.

Trypsin protease activity was analyzed by Amplite® Universal Fluorimetric Protease Activity Assay Kit. Protease substrate was incubated with 1 unit trypsin in the kit assay buffer. The control wells had protease substrate only (without trypsin). The fluorescence signal was measured starting from time 0 when trypsin was added. Samples were done in triplicates.

Protocol

SDS

COA

Catalog	Size	Price	Quantity
13500	500 Tests	Price

Citations

View all 12 citations: Citation Explorer

Genetically programmed engineered living materials as high-performance bioplastics

Authors:

Altin-Yavuzarslan, Gokce and Drake, Kinsey and Yuan, Shuo-Fu and Brooks, Sierra M and Kwa, Eng and Alper, Hal S and Nelson, Alshakim

Journal:

Matter (2024)

Autophagy and UPS pathway contribute to nicotine-induced protection effect in Parkinson’s disease

Authors:

Ullah, Inam and Uddin, Shahab and Zhao, Longhe and Wang, Xin and Li, Hongyu

Journal:

Experimental Brain Research (2024): 1--16

Comparative assessment of commercially available wound gels in ex vivo human skin reveals major differences in immune response-modulatory effects

Authors:

Seiser, S and Cerbu, D and Gallhofer, A and Matiasek, J and Elbe-B{\"u}rger, A

Journal:

Scientific reports (2022): 1--9

Optimization of Isolation Method for Extracellular Vesicles from Pancreatic Juice and Impact of Protease Activity

Authors:

Tsutsumi, Koichiro and Ueta, Eijiro and Kato, Hironari and Matsumoto, Kazuyuki and Horiguchi, Shigeru and Okada, Hiroyuki

Journal:

Digestive diseases and sciences (2022): 1--8

A novel polyethylene glycol (PEG)-drug conjugate of Venetoclax, a Bcl-2 inhibitor, for treatment of acute myeloid leukemia (AML)

Authors:

Ando, Hidenori and Murakami, Yuta and Eshima, Kiyoshi and Ishida, Tatsuhiro

Journal:

Cancer Reports (2021): e1485

References

View all 30 references: Citation Explorer

Transient kinetic experiments demonstrate the existence of a unique catalytic enzyme form in the peptide-stimulated ATPase mechanism of Escherichia coli Lon protease

Authors:

Vineyard D, Zhang X, Lee I.

Journal:

Biochemistry (2006): 11432

Highly stable glycosylated serine protease from the medicinal plant Euphorbia milii

Authors:

Yadav SC, P and e M, Jagannadham MV.

Journal:

Phytochemistry (2006): 1414

Effects of Pseudomonas fluorescens M3/6 bacterial protease on plasmin system and plasminogen activation

Authors:

Frohbieter KA, Ismail B, Nielsen SS, Hayes KD.

Journal:

J Dairy Sci (2005): 3392

Fibrillar amyloid beta-protein inhibits the activity of high molecular weight brain protease and trypsin

Authors:

Chauhan V, Sheikh AM, Chauhan A, Spivack WD, Fenko MD, Malik MN.

Journal:

J Alzheimers Dis (2005): 37

Characterization of a novel and specific inhibitor for the pro-apoptotic protease Omi/HtrA2

Authors:

Cilenti L, Lee Y, Hess S, Srinivasula S, Park KM, Junqueira D, Davis H, Bonventre JV, Alnemri ES, Zervos AS.

Journal:

J Biol Chem (2003): 11489

Spectral properties

Correction factor (280 nm)	0.35
Extinction coefficient (cm ^-1 M ^-1)	73000
Excitation (nm)	491
Emission (nm)	516
Quantum yield	0.92

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
UNSPSC	12352200

Instrument settings

Fluorescence microplate reader
Excitation	490 nm
Emission	525 nm
Cutoff	515 nm
Recommended plate	Solid black

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Page updated on October 8, 2025

Component A: Protease Substrate	1 vial (300 µL)
Component B: Trypsin	1 vial (100 µL, 5 U/µL)
Component C: 2X Assay Buffer	1 bottle (30 mL)