Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Cell Lysate, Tissue
Product nameGelatin Degradation Assay Kit (Alternative to Zymography)
Sample typeTissue, Cell Lysate
Species reactivityReacts with: Mammals, Other species
Gelatin Degradation Assay Kit (Alternative to Zymography) (ab234057) utilizes a hybrid approach for the detection of gelatinase activity by employing a highly quenched gelatin substrate which upon cleavage by a suitable gelatinase releases a fluorophore, which can be easily quantified using a conventional microplate reader. This method is substrate-specific, simple, fast, high-throughput adaptable and amenable to the sensitive detection of gelatinase activity (as low as 0.06 mCDU for bacterial collagenase) in biological samples.
This product is manufactured by BioVision, an Abcam company and was previously called K444 Gelatinase (Gelatin Degradation/Zymography) Assay Kit (Fluorometric). K444-100 is the same size as the 100 test size of ab234057.
Gelatinases are a type of matrix zinc-dependent metalloproteases (MMPs) that degrade gelatins and a variety of other extracellular matrix proteins. These enzymes are synthesized as latent zymogens that are activated by proteolysis and inhibited by tissue inhibitors of metalloproteases (TIMPs). Two mammalian gelatinases, Gelatinase A (MMP-2) and Gelatinase B (MMP-9), are critical for basement membrane degradation and are highly upregulated in variety of tumor cells. Gelatinase activity is usually detected by small peptide-based activity assays which may suffer from lack of substrate specificity. Other methods for gelatinase activity include gelatin Zymography where samples are electrophoresed on a gelatin-containing SDS-PAGE, and further renatured in a suitable buffer for 12-16 h. The zymogram is subsequently stained, and areas of digestion appear as clear bands against a darkly stained background where the substrate has been degraded by the enzyme. Such methods are laborious, time-consuming and may lead to the loss of enzymatic activity as renaturation may not be completely reversible.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 100 tests Cell Lysis Buffer 1 x 25ml Enzyme Positive Control 1 x 100µl FITC Standard (5 mM) 1 x 10µl Gelatinase Assay Buffer 1 x 25ml Gelatinase Substrate (FITC) 1 vial
RelevanceGelatin is denatured collagen composed of heterogeneous mixture of water-soluble proteins of high average molecular weights. Below 35-40°C gelatin swells and absorbs 5-10 times its weight of water to form a gel. Gel strength and viscosity gradually weaken upon prolonged heating in solution above 40°C.
ab234057 has been referenced in 1 publication.
- Khan A et al. Recombinant porcine NK-lysin inhibits the invasion of hepatocellular carcinoma cells in vitro. Int J Biol Macromol 140:1249-1259 (2019). PubMed: 31465800