Cathepsin H Inhibitor Screening Kit utilizes the ability of an active Cathepsin H to cleave a synthetic AFC-based peptide substrate to release free AFC, which can be easily quantified using a fluorometer or fluorescence microplate reader. In the presence of a Cathepsin H-specific inhibitor, the cleavage of this substrate is reduced/abolished resulting in decrease or total loss of the AFC fluorescence. This simple and high-throughput adaptable assay kit can be used to screen/study/characterize potential inhibitors of Cathepsin H.
Cathepsin H (CTSH, EC 18.104.22.168), a lysosomal cysteine protease, is important in the overall degradation of lysosomal proteins. It is composed of a dimer of disulfide-linked heavy and light chains, both produced from a single protein precursor. The encoded protein, which belongs to the peptidase C1 protein family, can act both as an aminopeptidase and as an endopeptidase. Increased expression of this gene has been correlated with malignant progression of prostate tumors.
Screen/study/characterize potential inhibitors of Cathepsin H
Store kit at -20°C, protected from light. Briefly centrifuge small vials at low speed prior to opening. Read the entire protocol before performing the experiment.CTSH Assay Buffer: Bring to room temperature before use. Store at 4°C or -20°C.CTSH Reagent: Aliquot & store at -20°C. Avoid repeated freeze/thaw.Human Cathepsin H: Reconstitute in 110 µl of CTSH Assay Buffer. Store at -20°C. Avoid repeated freeze/thaw. Use within two months.
CTSH Assay Buffer: 25 mlCTSH Reagent: 0.1 mlHuman Cathepsin H: 1 vialCTSH Substrate: 0.2 mlCTSH Inhibitor (1 mM): 20 µl
Fluorescence (Ex/Em = 400/505 nm)
Features & Benefits:
• Convenient & Rapid• High-throughput adaptable assay to screen/study/characterize the potential inhibitors of Cathepsin H• Cathepsin H inhibitor is included in the kit