||SensoLyte ® Rh110 Plasma Kallikrein Activity Assay kit *Fluorimetric* NEW
Human plasma kallikrein, a serine protease, is first synthesized as inactive plasma prekallikrein in liver and circulates in the plasma. Once activated by activated coagulation factor XII (FXIIa), plasma kallikrein releases bradykinin (an inflammatory mediator) from high molecular weight kininogen (HMWK).
The SensoLyte®Rh110 Plasma Kallikrein Activity Assay Kit employs a fluorescence peptide substrate for the detection of human plasma Kallikrein activity. This substrate contains rhodamine 110 fluorophore (Rh110). Plasma kallikrein cleaves this Rh110 substrate and results in the release of a bright green fluorescence and can be detected at excitation/emission=496 nm/520 nm. The longer wavelength spectra and higher extinction coefficient of Rh110 provide greater sensitivity and less interference from other reaction components. This assay can detect as low as 1 ng/mL active plasma Kallikrein. This kit could be used to detect enzyme activity in purified enzyme preparations, biological samples and it can also be applied for screening inhibitors.
Kit size: 100 assays (96-well plate)
|Features & Benefits|
|Detection of human plasma kallikrein|
|Compound screening for plasma kallikrein inhibitors|
|Fast & Convenient|
|The kit contains: |
| Plasma kallikrein substrate, Ex/Em=490/520nm upon cleavage|
|Human Plasma kallikrein |
|Plasma kallikrein inhibitor|
|Rh110 Fluorescence reference standard for calibration|
|Plasma prekallikrein activator|
|A detailed protocol|
Plasma Kallikerin detection
As an important vascular protease/biomarker, Plasma kallikrein represents the kallikrein-kinin system responsible for liberating bradykinin, a potent vasoactive pro-inflammatory peptide. It's detection in plasma lends an important indication of the cardiovascular state involving alternate and classical coagulation pathways.
Fig 1. Kallikrein activity present in plasma samples was detected by SensoLyte® Rh110 Plasma Kallikrein Activity Assay Kit. Plasma samples were diluted by 1X Assay buffer. The fluorescence signal was measured after 1 hour incubation of samples with substrate.
Fig 2. The specificity of SensoLyte® Rh110 Plasma Kallikrein Acitivty Assay Kit was tested with prekallikrein depleted human plasma. Lack of prekallikrein in the depleted plasma resulted in no conversion of prekallikrein to kallikrein.
Plasma Kallikrein Inhibitors Screening
Imbalance between plasma kallikrein and its naturally occurring plasma inhibitors is associated with disease states including hereditary angioedema (HAE), systemic lupus, inflammatory bowel disease and rheumatoid arthritis.
Our kit also provides an effective and convenient way to screen for plasma kallikrein inhibitors in your compound(s) of interest.
Fig 3. Inhibition of Kallikrein activity by PKSI-527 measured with SensoLyte® Rh110 Plasma Kallikrein Activity Assay Kit. Fluorescence signal was detected at Ex/Em=490/520nm (SpectraMax M5e).
More about plasma Kallikrein:
Plasma kallikrein is involved in several physiological and pathological processes such as blood coagulation, the classical complement cascade pathway and the activation of the alternative complement pathway1-5. Plasma kallikrein also plays a role in the induction of elastase release from neutrophils and conversion of pro-urokinase to urokinase in fibrinolysis6-9.
Material Safety Data Sheets (MSDS)
||Component C at -80 °C, Component A, B, D, E at -20 °C and Component A and B avoid light and moisture
||1. Bryant, J.W. et al, Cardiovasc Hematol Agents Med Chem 7, 234 (2009)
2. Toossi, Z. et al. Proc Natl Acad Sci U S A 89, 11969 (1992)
3. Ghebrehiwet, B. et al. J Exp Med 153, 665 (1981)
4. Qin, X.P. et al. Clin Exp Pharmacol Physiol 36, 319 (2009)
5. Phipps, J.A. et al. Hypertension 53, 175 (2009)
6. DiScipio, R.G. Immunology 45, 587 (1982)
7. Kaplan, A.P. et al. J Exp Med 135, 81 (1972)
8. Wachtfogel, Y.T. et al. J Clin Invest 72, 1672 (1983)
9. Ichinose, A. et al, J Biol Chem 261, 3486 (1986)