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Caspase / Apoptosis Detection

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Fluorescent Enzymatic Assays

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	Hydrogen Peroxide detection - Fluoro H₂0₂

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	Conjugate Diluents
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Cathepsin B,K,L Detection Kits

	Substrate based Assay to detect protease activity in hole living cells

Fluoro H₂0₂™
Fluorescent Hydrogen peroxide/Peroxidase Detection Kit

Key Benefits:

Quick 10 minute assay
Can monitor multiple time points to follow kinetics.
Dual mode, can detect H₂0₂ or peroxidase activity.
One-step, no wash assay.
Adaptable for High Throughput format
Non-destructive cell based assay allows monitoring of additional parameter
Applications - Fluorescent Plate Reader

Assay Principle

The Fluoro H₂0₂ detection kit utilizes a non-fluorescent detection reagent, to detect H₂0₂. H₂0₂ oxidizes the detection reagent in a 1:1 stoichiometry to produce a fluorescent product resorufin. This oxidation is catalyzed by Peroxidase in a homogeneous no wash assay system.

The detection reagent can be utilized to detect H₂0₂ release from cells or enzyme coupled reactions (1-7)

Reaction:

                                                          peroxidase
H₂0₂ + detection reagent (non-fluorescent) --------->   Resorufin
                                                                            (fluorescent)

Excitation: 530-571nm Emmission: 590-600nm

Citations

Deficiency of the Bax gene attenuates denervation-induced apoptosis - P. M. Siu and S. E. Alway - Apoptosis Journal Vol 11/No 6 June 2006

References

Anal Biochem 253, 162 (1997);
J. Immunol Methods 202, 133 (1997);
J Neurochem 79, 266 (2001);
Am J Physiol Lung Cell Mol Physiol 281, L993 (2001);
Mol Hum Reprod 7, 237 (2001);
Anal Biochem 287, 196 (2000);
J Invest Dermatol 112, 751 (1999).

Kit contents (for 500 assays)

5X Reaction Buffer: 20 ml buffer, pH 7.4
Detection reagent: One vial for 500 assays.
Hydrogen Peroxide: 200µL of a stabilized 3% solution.
Horseradish Peroxidase: 18.9 Units of enzyme

The following kits are available:
Catalog #	Size	Price (US$)
FLOH 100-3	500 Tests	$195

* Please call 888 7 ASSAYS (888-727-7297) or email info@celltechnology.com for volume pricing

Product Data Sheet

Protocol

MSDS

Figure 1. Hydrogen Peroxide titrated in 1X reaction Buffer and measured using the Fluoro H₂0₂ Kit.