
Product Name | SensoLyte ® Plus 520 MMP - 1 Assay Kit*Fluorimetric and Enhanced Selectivity* |
Size | 1 kit |
Catalog # | AS-72012 |
US$ | $601 |
The SensoLyte® Plus 520 MMP-1 Assay Kit is designed for specifically detecting MMP-1 activity in biological samples which may contain multiple MMPs, such as culture medium, serum, plasma, synovial fluid, and tissue homogenate. A specific anti-MMP-1 monoclonal antibody is used in combination with an MMP fluorogenic substrate, 5-FAM/QXL®520 FRET peptide. The fluorescence signal is monitored at Ex/Em=490 nm/520 nm upon MMP-1-induced cleavage of the FRET substrate. Ample materials are provided to perform 96 assays in a 96-well format. | |
Detailed Information | ![]() ![]() ![]() |
References | Jian J, Pelle E, Yang Q, Pernodet N, Maes D, Huang X. Iron sensitizes keratinocytes and fibroblasts to UVA-mediated matrix metalloproteinase-1 through TNF-? and ERK activation. Exp Dermatol. 2011 Mar; 20(3):249-54. PMID:20701626 |
Product Citations | Carrizzo, A. et al. (2015). Pentraxin 3 induces vascular endothelial dysfunction through a P-selectin/Matrix Metalloproteinase-1 pathway. Circulation 131, 1495. doi: 10.1161/CIRCULATIONAHA.114.014822.Atay, S. et al. (2014). Oncogenic KIT-containing exosomes increase gastrointestinal stromal tumor cell invasion. PNAS 111, 711. doi: 10.1073/pnas.1310501111.Halvorsen, B. et al. (2014). Increased levels of CCR7 ligands in carotid atherosclerosis: different effects in macrophages and smooth muscle cells. Cardiovasc Res 102, 148. doi: 10.1093/cvr/cvu036.Mogami, H. et al. (2014). Effect of thrombin on human amnion mesenchymal cells, mouse fetal membranes, and preterm birth. J Biol Chem 289, 13295. doi: 10.1074/jbc.M114.550541.Ozeki, N. et al. (2014). IL-1β-induced matrix metalloproteinase-13 is activated by a disintegrin and metalloprotease-28-regulated proliferation of human osteoblast-like cells. Exp Cell Res 323, 165.Mogami, H. et al. (2013) Fetal fibronectin signaling induces Matrix Metalloproteases and Cyclooxygenase-2(COX-2) in amnion cells and preterm birth in mice JBC 2883,1953 doi:10.1074/jbc.M112.424366Bogatkevich, GS. et al. (2012). The PPARγ agonist Rosiglitazone is antifibrotic for scleroderma lung fibroblasts: mechanisms of action and differential racial effects. Pulmonary Med doi:10.1155/2012/545172Leong, D. et al. (2011). Physiological loading of joints prevents cartilage degradation through CITED2. FASEB J 25, 182.Park, C. et al. (2011). Epidermal growth factor-induced matrix metalloproteinase-1 expression is negatively regulated by p38 MAPK in human skin fibroblasts. J Dermatol Sc 64, 134.Handy, J. et al. (2010). Adiponectin activation of AMPK disrupts leptin-mediated hepatic fibrosis via suppressors of cytokine signaling (SOCS-3). J Cell Biochem 110, 1195.Cowan, RW. et al. (2009). Collagenase expression and activity in the stromal cells from giant cell tumour of bone. Bone 44, 865.Jian, J. et al. (2011). Iron sensitizes keratinocytes and fibroblasts to UVA-mediated matrix metalloproteinase-1 through TNF-α and ERK activation. Exp Dermatol 20, 249.Moor, A. et al. (2009). Proteolytic activity in wound fluids and tissues derived from chronic venous leg ulcers. Wound Repair Regen 17, 832.Shi, Z-D. et al. (2009). Interstitial flow promotes vascular fibroblast, myofibroblast, and smooth muscle cell motility in 3-D collagen I via upregulation of MMP-1. Am J Physiol Heart Circ Physiol 10.1152/ajpheart.00369.2009.Qazi, H. et al. (2011). Fluid shear stress regulates the invasive potential of glioma cells via modulation of migatory activity and matrix metalloproteinase expression. PLoS One 6, e20348. |
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谁用过亲水性改性的聚丙烯膜养过细胞,最好是whatman的,效果怎么样?十分感激!:)
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新帖要有题目,且要尽量展示帖子主要内容意思~bywzmc225
本版发帖须知及新手指南
主药:50mg
mcc:20mg
l-hpc:10mg
cms-na:10mg
5%pvp50%乙醇qs
外加
ms:0.5mg
cms-na:3mg
50度干燥
以上处方不粘冲,但因上处方的辅料与主药有相互作用,导致片子变黄
现换成以下处方
处方1
主药:50mg
预胶化淀粉:20mg
淀粉:20mg
8%淀粉浆:qs
外加
ms:0.5或1mg
滑石粉:3或5mg
60度干燥
以上处方粘冲,为何?
处方2
主药:50mg
预胶化淀粉:20mg
淀粉:10mg
甘露醇:10mg
8%淀粉浆:qs
外加
ms:0.5或1mg
滑石粉:3或5mg
60度干燥
以上处方粘冲,为何
protocol如下:
灌注取脑后,4%多聚甲醛4℃固定24-48h。修块3-4mm。
脱水(常温):45%、55%、65%、75%、85%、95%、100%酒精各30min;
透明(常温):(100%酒精+二甲苯)/210min,二甲苯①20min,二甲苯②至完全透明(约20min)
浸蜡(60℃):(二甲苯+石蜡①)/210min,石蜡①30min,石蜡②40min;石蜡①为低熔点,石蜡②为高熔点
包埋:浸蜡后用石蜡②手动包埋;
切片厚度:6um、(8um也切的有)
展片:40℃水中展片;
烤片:60摄氏度烤片机侧烤约5min后吸去水分,转移至60℃温箱烤约4h;
脱蜡:烤片后趁蜡未凝固,进行脱蜡。二甲苯5min×2,100%酒精3min,95%酒精2min,
85%酒精2min,75%酒精2min,流水冲洗2min(放在缸子里用水泡的,换了几次水);
染色:
HE染色:苏木精5-8min(新配为5min,时间长可为8min,用前过滤),流水冲洗3min
1%盐酸酒精分化5s,流水冲洗1min,0.1%伊红1-2min,
脱水:85%酒精2min,95%酒精2min,100%酒精2min,
100%无水乙醇:二甲苯(1:1)2min
透明:二甲苯(I)3min,二甲苯(II)3min;
CV染色:浸入CV染液中10-20min,
脱水:75%乙醇2min,85%乙醇2min,95%乙醇2min,100%无水乙醇2min,
100%无水乙醇:二甲苯(1:1)2min
透明:二甲苯(I)3min,二甲苯(II)3min;
封片:中性树胶封片(整个染色过程避免干片)。
部分HE染色、CV染色片子结果如图所示,基本上没有完整的片子(染色时,片子随着在液体里的时间脱片、烂片的程度加深),另CV染色着色浅。
请教战友是什么原因?哪里出了问题,该怎么解决?
红色的是胰岛素,绿色的是胰高血糖素。都是石蜡切片~
请问下我这染出来的应该是胰岛细胞吧,背景颜色适合吗?
为什么胰高血糖素然不出来,这两种我用的方法是一样的。
方法:
1.脱蜡,水化:脱蜡前应将组织芯片在60摄氏恒温箱中烘烤1h
A:组织芯片置于二甲苯中浸泡15min,更换二甲苯后再浸泡10min(脱蜡)
B:无水乙醇中浸泡5min(水化)
C:95%乙醇中浸泡5min
D:75%乙醇中浸泡5min
2.PBS洗2次各5min
3.PBS洗2次每次5min,洗去枸橼酸钠(Ⅲ,Ⅱ)
4.3%H2O2(80%甲醇稀释)室温静置10min(去除内源性酶)
5.PBS洗2次各5min(Ⅰ、Ⅱ)
6.加入1%BSA稀释(0.01MPBS稀释)封闭液,37度恒温烘箱1h,顷去多余液体,不洗
7.滴加一抗用1%BSA稀释(1:50)或(1:100),先室温放置1h,然后4度过夜(放入湿盒)并用PBS代替一抗做阴性对照
8.PBS洗3次每次5min
9.滴加二抗(1:100)或(1:200),用1%BSA稀释37度恒温箱(放入湿盒)1h,避光
10.PBS洗3次各2min,避光,用摇床洗涤
11.加入20ul样品Hoechst液体(10ug/ml)5min用摇床避光
12.PBS洗3次各2min,避光,用摇床洗涤
13.滴加抗荧光猝灭剂,封片,镜检,绿色荧光
尤其是前面两个,后面的还可以从字面理解着做,前面这两个就纯把我搞晕菜了.
全量法:测水浸出物的
酒石酸铁比色法;
紫外分光光度法;
恒重法:测水含量的
谢谢谢谢

