Pepsin/Pepsinogen Assay Kit (Fluorometric) (ab239722)
Key features and details
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Other biological fluids, Plasma, Serum
Overview
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Product name
Pepsin/Pepsinogen Assay Kit (Fluorometric) -
Detection method
Fluorescent -
Sample type
Serum, Plasma, Other biological fluids -
Product overview
Pepsin/Pepsinogen Assay Kit (Fluorometric) (ab238722) is a homogenous assay that allows for quantification of pepsin activity in gastric tissues (stomach, duodenum etc.) and various biological fluids (serum/plasma, gastric juice, vomit). The assay utilizes a synthetic peptide substrate bearing both a fluorophore and a fluorescence quencher. Upon cleavage by pepsin, the fluorophore-bearing peptide fragment is unquenched to produce a bright fluorescent signal (Ex/Em = 328/418 nm). Lysosomal aspartic proteases in the peptidase A1 family (Cathepsin D and E) do not interfere with the assay. The assay is rapid, simple to perform and is vastly more sensitive than the classical hemoglobin degradation assay, with a detection limit of 500 μU pepsin activity per well.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components Identifier 100 tests Mca Standard (1mM) Yellow 1 x 25µl Pepsin Assay Buffer 1 x 25ml Pepsin Inhibitor (Pepstatin A) 1 x 20µl Pepsin Positive Control 1 vial Pepsin Substrate 1 x 200µl -
Research areas
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Relevance
Pepsin is a digestive enzyme with particularly broad specificity. It is stored as pepsinogen, so only released when needed. In the stomach, chief cells release pepsinogen. The hormone gastrin and the vagus nerve trigger the release of both pepsinogen and HCl from the stomach lining when food is ingested. HCl creates an acidic environment which allows pepsinogen to unfold and cleave itself in an autocatalytic fashion, thereby generating pepsin (the active form). Pepsin may then cleave the dietary proteins into smaller peptides. -
Cellular localization
Secreted -
Alternative names
- Pepsin
- Pepsin A
- Pepsin A 3
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Images
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MCA standard curve
One mole of MCA corresponds to the cleavage of one mole of fluorogenic Pepsin Substrate
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Kinetics of Pepsin Substrate metabolism by porcine gastric mucosal pepsin (3.33 ng purified enzyme) and specificity of substrate metabolism by pepsin versus other aspartic proteases.The acid-activated proteases Cathepsin D and E exhibit minimal assay interference, even when present at ≥300-fold excess by mass.
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Estimation of pepsinogen activity in pooled normal human serum and single-donor serum from a gastric ulcer patient with confirmed H. Pylori infection (each 10 µl of undiluted serum).Data are mean ± SEM of 3 replicates, assayed according to the kit protocol.