Gelatin Zymography/Fluorescent Labeled

Gelatin Zymographygrapzy Electrophoresis Kit

Simplifies the detection and confirmation of MMPs (matrix metalloproteinases) as much as possible

This is a kit for easily performing gelatin Zymography electrophoresis for detection and confirmation of MMPs contained in blood, body fluids, secretions, tissues, cells, and cell culture solutions of humans and various animals.

Zymography Kit Q&A

Features

MMPs (matrix metalloproteinases) are one of the metalloprotease families, and more than 20 types have been identified as enzymes involved in the metabolism of extracellular matrix components in vivo, such as tumor invasion/metastasis, inflammatory tissue, and angiogenesis. increase. As one of the detection methods of MMPs, electrophoretic separation and enzymatic activity detection (Zymography) are widely used. This kit consists of precast gels for gelatin Zymography using gelatin as a substrate, various buffers, and MMP markers. Please use it to detect and confirm MMP contained in blood, body fluids, secretions, tissues, cells, and cell culture solutions of humans and various animals.

The Fluorescent Labeled Gelatin Zymography Electrophoresis Kit consists of a precast gel with fluorescently labeled gelatin as a substrate protein, various buffers, and a set of MMP markers to facilitate gelatin Zymography electrophoresis. Fluorescently labeled gels allow continuous confirmation of enzymatic reactions.

	MMP activity in tear fluid by gelatin Zymography (left) Gelatin Zymography electrophoresis kit (Cosmo Bio-type, product number: AK47 is used) ( 　　　1) Pro-MMP9 (2) Pro-MMP2 (3) MMP2 (right) Electrophoresis pattern after time (using Cosmo Bio type, product number: AK83) 　　　M1: Molecular weight marker M2: MMP marker Photographed with a gel imager (fluorescence wavelength is 535 nm). Or refer to the FITC wavelength (excitation wavelength 495 nm, fluorescence wavelength 520 nm).

Table 1. Gelatin Zymography: comparison of unlabeled and fluorescently labeled

	unlabeled	fluorescent label
electrophoresis	1 hour	1 hour
Gel washing (SDS removal)	1 hour	1 hour
enzyme reaction	20 hours to 40 hours	3 hours to 40 hours
Visualization method	Protein staining 1 hour	Capable of continuous observation even during enzymatic reaction
bleaching	Several days	unnecessary

Composition

Atto type

• Compatible with Atto's mini gel type.

sign	Product name	Composition		Catalog number
Unlabeled	Gelatin Zymography electrophoresis kit (ATTO type)	12 samples x 5 times Enzyme reaction buffer (10x concentrated) 25 mL 1 Gelatin Zymographyphoresis precast gel 12 well 5 sheets (sold separately, part number: AK46) Electrophoresis buffer (10x concentrated) 100 mL 2 Washing solution (10x concentrated) 100 mL 1 Sample preparation buffer 5 mL 1 Staining solution 100 mL 1 MMP marker (ProMMP-2, MMP-2, ProMMP-9) 0.2 mL 1 vial		PMC-AK45
	Gelatin Zymography electrophoresis Precast gel (Atto type)	Gelatin Zymographyphoresis precast gel	5 sheets	PMC-AK46
		120W x 100H x 1 mm 12 well
		(Product number: Same as included in AK45)	10 sheets	PMC-AK49
Fluorescent label	Fluorescent Labeled Gelatin Zymography Electrophoresis Kit (Atto Type)	For 12 samples x 5 times Buffer for electrophoresis (10-fold concentrated) 100 mL 2 Washing buffer (10-fold Fluorescent-labeled gelatin Zymography electrophoresis precast gel 12 wells 5 sheets (sold separately, product number: AK81) concentrated) 100 mL 1 For this enzyme reaction Buffer (10x concentrated) 25 mL 1 MMP markers Sample conditioning buffer 5 mL 1 (ProMMP-2, MMP-2, ProMMP-9) 0.2 mL 1		PMC-AK80
	Fluorescent-labeled gelatin Zymography electrophoresis Precast gel (Atto type)	Gelatin Zymographyphoresis precast gel  120W x 100H x 1 mm 12 well 5 sheets	5 sheets	PMC-AK81
		(Product number: Same as included in AK80)	10 sheets	PMC-AK82

Cosmo Bio type

• Compatible with Cosmo Bio, Daiichi Kagaku, Novex, Biocraft, Tefco, Nippon Aid, and Marisol electrophoresis equipment.

sign	Product name	Composition		Catalog number
Unlabeled	Gelatin Zymography Electrophoresis Kit (Cosmo Biotype)	12 samples x 5 times Gelatin Zymographyphoresis precast gel 12 well 5 sheets (sold separately, part number: AK48) Electrophoresis buffer (10x concentrated) 100 mL 2 Washing solution (10x concentrated) 100 mL 1 Enzyme reaction buffer (10x concentrated) 25 mL 1 Sample preparation buffer 5 mL 1 Staining solution 100 mL 1 MMP marker (ProMMP-2, MMP-2, ProMMP-9) 0.2 mL 1 vial		PMC-AK47
	Gelatin Zymography electrophoresis precast gel (Cosmo Bio type)	Gelatin Zymographyphoresis precast gel	5 sheets	PMC-AK48
		100W x 100H x 1 mm 12 well
		(Product number: Same as included in AK47)	10 sheets	PMC-AK50
Fluorescent label	Fluorescent Labeled Gelatin Zymography Electrophoresis Kit (Cosmo Biotype)	12 samples x 5 times Fluorescent-labeled gelatin Zymography electrophoresis precast gel 12 wells 5 sheets (sold separately, product number: AK84) Buffer for electrophoresis (10-fold concentrated) 100 mL 2 Washing buffer (10-fold concentrated) 100 mL 1 For this enzyme reaction Buffer (10x concentrated) 25 mL 1 Sample conditioning buffer 5 mL 1 MMP markers (ProMMP-2, MMP-2, ProMMP-9) 0.2 mL 1		PMC-AK83
	Fluorescent-labeled gelatin Zymography electrophoresis Precast gel (Cosmo Bio type)	Gelatin Zymographyphoresis precast gel	5 sheets	PMC-AK84
		100W x 100H x 1 mm 12 well 5 sheets
		(Product number: Same as included in AK83)	10 sheets	PMC-AK85

Scale illustration of a precast gel plate

References

1. Miyazaki, K., Ohta, Y., Nagai, M., Morimoto, N., Kurata, T., Takehisa, Y., Ikeda, Y., Matsuura, T., Abe, K.
   Disruption of Neurovascular Unit Prior to Motor Neuron Degeneration in Amyotrophic Lateral Sclerosis.
   J. Neurosci. Res. 89, 718-728 (2011), (PMID:21337372)
2. Fujiwara, M., Kashima, T. G., Kunita, A., Kii, I., Komura, D., Grigoriadis, A. E., Kudo, A., Aburatani, H., Fukayama, M.
   Stable Knockdown of S100A4 Suppresses Cell Migration and Metastasis of Osteosarcoma.
   Tumour Biol. 32, 611-622 (2011), (PMID:21360024)
3. Aoki, T., Kataoka, H., Ishibashi, R., Nozaki, K., Hashimoto, N.
   Simvastatin Suppresses the Progression of Experimentally Induced Cerebral Aneurysms in Rats.
   Stroke. 39, 1276-85 (2008), (PMID:18309148)
4. Kuramochi, D., Unoki, H., Bujo, H., Kubota, Y., Jiang, M., Rikihisa, N., Udagawa, A., Yoshimoto, S., Ichinose, M.,Saito, Y.
   Matrix Metalloproteinase 2 Improves The Transplanted Adipocyte Survival in Mice.
   Eur. J. Clin. Invest. 38, 752-759 (2008), (PMID:18837800)
5. Fujimoto, M., Takagi, Y., Aoki, T., Hayase, M., Marumo, T., Gomi, M., Nishimura, M., Kataoka, H., Hashimoto, N., Nozaki, K.
   Tissue Inhibitor of Metalloproteinases Protect Blood-brain Barrier Disruption in Focal Cerebral Ischemia.
   J. Cereb. Blood Flow Metab. 28, 1674-1685 (2008), (PMID:18560439)
6. Nakano, K., Hokamura, K., Taniguchi, N., Wada, K., Kudo, C., Nomura, R., Kojima, A., Naka, S., Muranaka, Y., Thura, M., Nakajima, A., Masuda, K., Nakagawa, I., Speziale, P., Shimada, N., Amano, A., Kamisaki, Y., Tanaka, T., Umemura, K., Ooshima, T.
   The collagen-binding protein of Streptococcus mutans is involved in haemorrhagic stroke.
   Nat. Commun. 2, 485 (2011), (PMID:21952219)
7. Tahara, S., Nojima, S., Ohshima, K., Hori, Y., Kurashige, M., Wada, N., Ikeda, J., Morii, E.
   S100A4 accelerates the proliferation and invasion of endometrioid carcinoma and is associated with the "MELF" pattern.
   Cancer Sci. 107, 1345-52 (2016), (PMID:27348205)
8. Lourbakos, A., Yau, N., de Bruijn, P., Hiller, M., Kozaczynska, K., Jean-Baptiste, R., Reza, M., Wolterbeek, R., Koeks, Z., Ayoglu, B., de Klerk, D., Campion, G., Zaharieva, I., Nadarajah, VD., Nilsson, P., Al-Khalili Szigyarto, C., Muntoni, F., Lochmuller, H., Verschuuren, JJ., Goemans, N., Tulinius, M., Niks, EH., de Kimpe, S., Aartsma-Rus, A., 't Hoen, PAC., Spitali, P.
   Evaluation of serum MMP-9 as predictive biomarker for antisense therapy in Duchenne.
   Sci. Rep. 7, 17888 (2017), (PMID:29263366)

Gelatin Zymography electrophoresis kit (ATTO type)

Catalog Number	Product Name	Size
PMC-AK45	Gelatin Zymography Kit (ATTO type: wide thick plates)	1 Kit
PMC-AK45-001	Gelatin Zymography Reagent Kit (Cosmo Bio type: narrow thin plates)	1 Kit

Gelatin Zymography Electrophoresis Kit (Cosmo Bio type)

Catalog Number	Product Name	Size
PMC-AK47	Gelatin Zymography Kit (Cosmo Bio type: narrow thin plates)	1 Kit
PMC-AK48	Gelatin Zymography Precast Gels (Cosmo Bio type: narrow thin plates)	5 Plates
PMC-AK48	Gelatin Zymography Precast Gels (Cosmo Bio type: narrow thin plates)	10 Plates
PMC-AK83	FITC-labeled Gelatin Zymography Kit (Cosmo Bio type: narrow thin plates)	1 Kit
PMC-AK84	FITC-labeled Gelatin Zymography Precast Gels (Cosmo Bio type: narrow thin plates)	5 Plates
PMC-AK84	FITC-labeled Gelatin Zymography Precast Gels (Cosmo Bio type: narrow thin plates)	10 Plates

MMP marker

Catalog Number	Product Name	Size
PMC-AK38	Human MMP Marker	200 ul