FITC-labeled Gelatin Zymography Kit (Cosmo Bio type: narrow thin plates)

Cosmo Bio

Catalog No.: PMC-AK83

Product Description

Matrix metalloproteinases (MMPs) belong to the family of metalloproteinases, which consists of at least 20 members, and known to be involved in the metabolism of extracellular matrix proteins. MMPs are widely detected by zymography. The FITC-labeled Gelatin-Zymography Kit (Cat.No.PMC-AK83-COS) provides an easy system of the electrophoresis for zymography. This product is used for detecting ProMMP-2, MMP-2 and ProMMP-9 in blood, body fluid, secretion, cell lysate, cell culture medium, and other samples.

Citations for FITC-labeled Gelatin Zymography Kit (Cosmo Bio type: narrow thin plates) – 6 Found

Takagi, Yuya; Aoshima, Kota; Kuse, Yoshiki; Nakamura, Shinsuke; Okada, Yasunori; Shimazawa, Masamitsu. The Involvement of HYBID in the Regulation of Intraocular Pressure and Extracellular Matrix Accumulation in the Trabecular Meshwork. Biological & Pharmaceutical Bulletin. 48(10):1555-1565. 

Hoang, Thi Xoan; Dang, Nhat Minh; Bae, Kang Gyu; Kim, Jae Young. Anti-Inflammatory and Antioxidant Effects of Topical Formulations Containing Plant Extracts, Methylsulfonylmethane, and Peptiskin(®) in In Vitro Models of Arthritis. Pharmaceuticals (Basel, Switzerland). 2025;18(9) 

Ishida, Yuko; Kuninaka, Yumi; Komori, Tadasuke; Iwabuchi, Sadahiro; Nosaka, Mizuho; Kimura, Akihiko; Hashimoto, Shinichi; Morikawa, Yoshihiro; Mukaida, Naofumi; Kondo, Toshikazu. OSMRβ-mediated signals on resident fibroblasts restore healing in diabetic skin wounds through promoting angiogenesis and granulation tissue formation. Communications Biology. 2025;8(1):1505. 

Ogawa, Hisataka; Ekawa, Tomoya; Mizote, Yu; Akazawa, Takashi; Tahara, Hideaki. Relaxin-2-secreting CAR-T cells exhibit enhanced efficacy in stromal-rich xenograft tumors. Frontiers In Immunology. 16:1506204. 

Choudhury, Mohammed E; Ozaki, Saya; Miyaue, Noriyuki; Matsuura, Taisei; Mikami, Kanta; Islam, Afsana; Kubo, Madoka; Ando, Rina; Yano, Hajime; Kunieda, Takeharu; Nagai, Masahiro; Tanaka, Junya. Chloride Intracellular Channel Protein 2 Promotes Microglial Invasion: A Link to Microgliosis in the Parkinson's Disease Brain. Brain Sciences. 2022;13(1) 

Shimada, Yoshiaki; Shimura, Hideki; Tanaka, Ryota; Yamashiro, Kazuo; Koike, Masato; Uchiyama, Yasuo; Urabe, Takao; Hattori, Nobutaka. Phosphorylated recombinant HSP27 protects the brain and attenuates blood-brain barrier disruption following stroke in mice receiving intravenous tissue-plasminogen activator. Plos One. 13(5):e0198039. 

FITC-labeled Gelatin Zymography Kit (Cosmo Bio type: narrow thin plates) FAQs

What is zymography?
Zymography is a method for detecting enzymes based on their enzymatic activity after separation by electrophoresis. When a gel containing a substrate protein such as gelatin or casein is used, the entire gel is stained blue after protein staining. Regions with enzymatic activity appear as clear bands because the substrate protein has been degraded.

Why can proMMPs be detected as bands in zymography even though they are inactive enzymes? Why do proMMPs and MMPs have different molecular weights?
proMMPs are inactive because their active sites are masked by a propeptide sequence. During zymography, SDS treatment releases the propeptide sequence masking the active site, allowing even proMMPs to exhibit enzymatic activity. MMPs are activated when the propeptide region is cleaved from proMMPs. Therefore, precursor proMMPs have a higher molecular weight, whereas activated MMPs have a lower molecular weight.
For more information, please refer to the following publication:
Possible role of matrix metalloproteinases in reconstruction of peripheral pulmonary arteries induced by hypoxia. J. Novotná and J. Herget Physiol Res. 2002;51(4):323-334

What is the gel concentration of the precast gel included in Gelatin Zymography Kit (Cat. No. PMC-AK47) and FITC-labeled Gelatin Zymography Kit (Cat. No. PMC-AK83)?
The acrylamide concentration is 10%, and the gel contains 0.1% gelatin.

What are the dimensions of the precast gel included in Gelatin Zymography Kit (Cat. No. PMC-AK47) and FITC-labeled Gelatin Zymography Kit (Cat. No. PMC-AK83)?
Gel size: 100 × 100 mm
Gel thickness: 1 mm
Overall thickness including glass plates: 3.6 mm

How long does electrophoresis take at 15 mA when using Gelatin Zymography Kit (Cat. No. PMC-AK47) and FITC-labeled Gelatin Zymography Kit (Cat. No. PMC-AK83)?
Depending on the electrophoresis apparatus used, electrophoresis typically takes approximately 3 hours.

How can I obtain clear images of zymography gels?
For optimal visualization of bands, it is recommended to illuminate the gel from below using white light.

How much protein should be loaded per lane when using Gelatin Zymography Kit (Cat. No. PMC-AK47) and FITC-labeled Gelatin Zymography Kit (Cat. No. PMC-AK83)?
Since MMP activity varies depending on the tissue or cell type, it is difficult to define a universally optimal amount of protein to load. As an initial guideline, load approximately 10 µg of protein and determine the optimal amount based on the results obtained. When culture supernatants are used as samples, the volume of the sample is often used as the loading criterion.

How can MMPs be extracted from tissues?
Since MMPs are tightly associated with extracellular matrix components in tissues, extraction efficiency is generally low. One method is to homogenize tissue using a Polytron homogenizer in a buffer containing 10 mM CaCl₂ and 0.25% Triton X-100, followed by centrifugation. The resulting supernatant can then be used as the sample. Other extraction methods include urea solubilization and heat treatment at 60°C. Additional information is available in Woessner Jr., Methods in Enzymology, Vol. 248, pp. 510-529.

What should be avoided during sample preparation for Gelatin Zymography Kit (Cat. No. PMC-AK47) and FITC-labeled Gelatin Zymography Kit (Cat. No. PMC-AK83)?
The use of reducing agents such as 2-mercaptoethanol and dithiothreitol (DTT), as well as heat treatment, should be avoided because they may denature proteins and result in loss of enzymatic activity.

Is there any information regarding the effects of blood collection anticoagulants on samples used with Gelatin Zymography Kit (Cat. No. PMC-AK47) and FITC-labeled Gelatin Zymography Kit (Cat. No. PMC-AK83)?
Gerlach et al. (Analytical Biochemistry, Vol. 344, pp. 147-149) reported a comparison of serum and plasma prepared using three anticoagulants: citrate, heparin, and EDTA. Please refer to the publication for details.

What is the band detected between proMMP2 and MMP2 when using Gelatin Zymography Kit (Cat. No. PMC-AK47) and FITC-labeled Gelatin Zymography Kit (Cat. No. PMC-AK83)?
The band corresponds to intermediate MMP2. MMP2 is activated by cleavage of the propeptide region from proMMP2, and intermediate MMP2 represents a processing intermediate that appears during this activation process.

Is the mobility of proMMP9 different between mice and humans when using Gelatin Zymography Kit (Cat. No. PMC-AK47) and FITC-labeled Gelatin Zymography Kit (Cat. No. PMC-AK83)?
Yes. MMP9 contains both N-linked and O-linked glycans, and the size of these glycans differs among animal species. Therefore, mouse-derived proMMP9 is detected at a higher molecular weight than human-derived proMMP9.

What could the bands above proMMP9 represent when using Gelatin Zymography Kit (Cat. No. PMC-AK47) and FITC-labeled Gelatin Zymography Kit (Cat. No. PMC-AK83)?
The bands may represent MMP9 dimers or complexes formed between MMP9 and other proteins, such as TIMPs, lipocalin, or α2-macroglobulin.