明胶酶谱试剂盒（COSMOBIO） Gelatin Zymography Kit(COSMOBIO)

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明胶酶谱试剂盒（COSMOBIO） Gelatin Zymography Kit(COSMOBIO)明胶酶谱试剂盒

Gelatin Zymography Kit



◆背景


基质金属蛋白酶（MMP）属于金属蛋白酶家族，由至少20个成员组成，MMP参与细胞外基质蛋白的代谢。MMP常用酶谱法检测。明胶酶谱试剂盒（Cat.No.AK47-COS和AK45-COS）提供了用于酶谱电泳的简单系统。该产品用于检测血液，体液，分泌物，细胞裂解液，细胞培养基和其他样品中的ProMMP-2，MMP-2和ProMMP-9。

明胶酶谱试剂盒（COSMOBIO） Gelatin Zymography Kit(COSMOBIO)

◆试剂盒组成


组成

包装

保存

Precast gel for 12 samples

5 pieces

4°C

电泳缓冲液 (×10)

100 mL × 2

4°C

洗涤缓冲液 (×10)

100 mL × 1

4°C

反应缓冲液 (×10)

25 mL × 1

4°C

样品制备缓冲液

5 mL × 1

4°C

染色液

100 mL × 1

室温

MMP markers (ProMMP-2, MMP-2, ProMMP-9)

0.2 mL

4°C


保质期：自生产日期起6个月。

凝胶规格：

COSMO BIO 型：玻璃板凝胶大小是100(W)×100(H)，厚度为1 mm.
　　　　　         1个试剂盒中包含60个样品量的试剂。
ATTO 型：玻璃板凝胶大小是120(W)×100(H)，厚度为1 mm.
　　　　　1个试剂盒中包含60个样品量的试剂。


该试剂盒中的预制胶与以下产品兼容：

　　Life technologies: Novex
　　BIO CRAFT: 电泳系统等

◆应用实例

明胶酶谱试剂盒（COSMOBIO） Gelatin Zymography Kit(COSMOBIO)

Fig.1 上样10 μL MMP marker在37℃下酶反应24小时。


 

相关资料


明胶酶谱试剂盒（COSMOBIO） Gelatin Zymography Kit(COSMOBIO)

欲了解详情请点击图片


◆操作步骤


1.  Load 100-150 mL of Electrophoresis Buffer to the lower chamber (anode). Refer to the instruction manual of your electrophoresis tank/chamber to know appropriate volume of the   buffer.
2.  Take out the comb on precast gel carefully and set the gel to the electrophoresis chamber. The side of the sample holes should be set on the upper side. If the sample holes are disturbed, fix them up with a needle etc.
3.  Load approximately 100 mL of Electrophoresis Buffer to the upper chamber (cathode).
4.  Mix samples with equivalent volume of sample preparation buffer. Incubate for 15 minutes at Room Temperature. (Do NOT heat the samples.)
5.  Load approximately 100 mL of Electrophoresis Buffer to the upper chamber (cathode).
6. 

Apply the samples and MMP markers to gel plate. (Fig.1: 10ml of MMP markers applied and enzymatic reacted for 24 hrs. at 37°C)  Run electrophoresis at 15 mA constant current. (If you use 2 gels, set the current at 30 mA)

*MMP markers can be used without the sample preparation buffer. 
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 the gel 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 the 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 30minutes several hours to de-stain.

◆细胞分析相关研究用试剂盒


产品编号 产品名称 简介

PMC-AK04F-COS

TRAP Staining Kit

抗酒石酸酸性磷酸酶染色试剂盒（TRAP）

TRAP染色试剂盒用于破骨细胞中酒石酸盐抗性酸性磷酸酶的染色试剂盒。

PMC-AK02-COS

Stains All Gel Staining Kit for Acidic Proteins

酸性蛋白凝胶染色试剂盒

该试剂盒专门用于染色调节骨钙化的强酸性蛋白质，如骨钙素，骨桥蛋白和BSP II。

PMC-AK19F-COS

Adipocyte Fluorescent Staining Kit

脂肪细胞荧光染色试剂盒

用Bodipy染色和H33258的细胞核荧光染色细胞中的脂肪球。

PMC-AK11F-COS

Pancreatic beta-cell Fluorescent Staining Kit

胰腺β细胞染色试剂盒

PMC-AK03-COS

Acidic Mucopolysaccaride Assay Kit

酸性 黏多糖检测试剂盒

PMC-AK06-COS

DNA Quantity Kit

DNA定量试剂盒

无需纯化可从培养细胞定量DNA。

PMC-AK37-COS

TypeI collagenase assay kit I

型胶原酶检测试剂盒

PMC-AK09F-COS

Lipid Assay Kit

血脂检测试剂盒

该试剂盒（油红O亲脂红色染料）染色细胞内油滴，并能够用有机溶剂萃取法定量。

PMC-AK01-COS

GPDH Assay Kit

甘油磷酸脱氢酶检测试剂盒

与常规方法相比，该试剂盒在测量酶活性方面具有数个优点，包括更高的稳定性和再现性。

PMC-AK20-COS

Alkaline Phosphatase Staining Kit

碱性磷酸酶染色试剂盒

用于成骨细胞中碱性磷酸酶的染色。

PMC-AK21-COS

Calcified nodule Staining kit

钙化结节染色试剂盒

PMC-AK12-COS

POLARIC

活细胞溶致变色荧光

PMC-AK47-COS

Gelatin zymography kit

明胶酶谱试剂盒（COSMOBIO）

PMC-AK38-COS

MMP Marker

MMP标记

参考文献


[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)
[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 CellMigration and Metastasis of Osteosarcoma. Tumour Biol.
[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)
[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)
[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)
仅用于研究。不得用于人类基因治疗或临床诊断。

产品编号 产品名称 产品规格 产品等级
PMC-AK47-COS 明胶酶谱试剂盒（COSMOBIO） 
Gelatin Zymography Kit (Cosmo Bio type)		 1 KIT －
PMC-AK48-COS 预制明胶酶谱凝胶（COSMOBIO） 
Precast Gelatin Zymogram Gel (Cosmo Bio type)		 5PLATE －
PMC-AK50-COS 预制明胶酶谱凝胶（COSMOBIO） 
Precast Gelatin Zymogram Gel (Cosmo Bio type)		 10PLATE －
PMC-AK38-COS MMP标记 
MMP Marker		 200 UL －
PMC-AK45-COS 明胶酶谱试剂盒（ATO） 
Gelatin zymography kit (ATTO type)		 1 KIT －
PMC-AK46-COS 预制明胶酶谱凝胶（ATO） 
Precast Gelatin Zymogram Gel (ATTO type)		 5PLATE －
PMC-AK49-COS 预制明胶酶谱凝胶（ATO） 
Precast Gelatin Zymogram Gel (ATTO type)		 10PLATE －