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

Cosmo Bio LTD

Catalog No: PMC-AK47

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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 Gelatin-Zymography Kit (Cat.No.AK47-COS and AK45-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.

Our Zymography precast gels are available in two types (sizes). Please choose the type that fits your gel box. 
     ATTO type: Glass plate gel is 120(W)×100(H) and the thickness is 1mm
     Cosmo Bio type: Glass plate gel is 100(W)×100(H) and the thickness is 1mm.  

Zymography Kit 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. [Cautions: This product has a short expiration date (less than 3 months). Please use it after arrival as soon as possible.

Protocol:

1. Load 100-150 mL of Electrophoresis Buffer to the lower chamber (anode). Refer to the instruction manual ofyour electrophoresis tank/chamber to know appropriate volume of the buffer.
2. Take out the comb on precast gel carefully and set the gel to electrophoresis chamber. The side of sample holes should be set on the upper side. If the sample holes aredisturbed, fix them up with needle etc.
3. Load around 100 mL of Electrophoresis Buffer to the upper chamber (cathode).
4. Mixsamples with equivalent volume of sample preparation buffer. Incubate for 15 minutes at Room Temperature. (Do NOT heat thesamples.)
*MMP markers can be used without sample preparation buffer.
5. Apply the samples and MMP markers to gel plate.
6.Run electrophoresis at 15 mA constant current. (If you use 2 gels, set the current at 30 mA)
7. After the run is completed, turn off the electrophoresis chamber, and take out the gel plate from the chamber.
8. Remove the upper glass plate of thegel plate and peel off the gel carefully with a spatula.
9. Put the gel in the tray with 200 ml of Washing Buffer. Incubate with shaking at Room Temperature.
10. Put the gel in the container with 50 ml of Reaction Buffer and seal up the container. Incubate the gel in the container in incubator at 37°C for 20-40 hrs (Lower enzyme concentration needs longer reaction time.).
11. After enzymatic reaction, put the gel in the container with Staining solution. Incubate for 30 minutes at Room Temperature to stain protein.
12. Put the gel in the container with De-staining solution. And incubate for 30 minutes to several hours to de-stain.

Gelatin Zymography Kit 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. [Cautions: This product has a short expiration date (less than 3 months). Please use it after arrival as soon as possible.

 

 

Citations

• Jong Youl Kim, Nuri Kim, Zhen Zheng, Jong Eun Lee, and Midori A. Yenari
  The 70kD heat shock protein protects against experimental traumatic brain injury
  Neurobiol Dis. 2013 October ; 58: 289–295. PMID: 23816752
• Yang Zheng, Seiji Yamamoto, Yoko Ishii, Yang Sang, Takeru Hamashima, Nguyen Van De, Hirofumi Nishizono, Ran Inoue, Hisashi Mori, and Masakiyo Sasahara
  Glioma-Derived Platelet-Derived Growth Factor-BB Recruits Oligodendrocyte Progenitor Cells via Platelet-Derived Growth Factor Receptor-a and Remodels Cancer Stroma
  The American Journal of Pathology, Vol. 186, No. 5, May 2016 PMID: 26945107

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References

• 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
• 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. 2011 Jun;32(3):611-22.
  PMID: 21360024
• 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
• 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
• 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

Citations for Gelatin Zymography Kit (Cosmo Bio type: narrow thin plates) – 5 Found

Galectin-4 expression is down-regulated in response to autophagy during differentiation of rat trophoblast cells.
Sci Rep. 2016 Aug 30;6:32248. doi: 10.1038/srep32248. 

Toll-like receptor 9 prevents cardiac rupture after myocardial infarction in mice independently of inflammation.
Am J Physiol Heart Circ Physiol. 2016 Dec 1;311(6):H1485-H1497. doi: 10.1152/ajpheart.00481.2016. Epub 2016 Oct 21. 

White Matter Injury After Subarachnoid Hemorrhage: Role of Blood-Brain Barrier Disruption and Matrix Metalloproteinase-9.
Stroke. 2015 Oct;46(10):2909-15. doi: 10.1161/STROKEAHA.115.010351. Epub 2015 Sep 15. 

Cardiac-specific ablation of the STAT3 gene in the subacute phase of myocardial infarction exacerbated cardiac remodeling.
Am J Physiol Heart Circ Physiol. 2015 Aug 1;309(3):H471-80. doi: 10.1152/ajpheart.00730.2014. Epub 2015 Jun 8. 

The Role of Oxidative Stress in Microvascular Disturbances after Experimental Subarachnoid Hemorrhage.
Transl Stroke Res. 2019 Dec;10(6):684-694. doi: 10.1007/s12975-018-0685-0. Epub 2019 Jan 9.