
Product Name | SensoLyte ® AFC Plasmin Activity Assay Kit *Fluorimetric* |
Size | 1 kit |
Catalog # | AS-72124 |
US$ | $380 |
Plasmin belongs to the family of serine proteases. It plays a key role in fibrinolysis by dissolving fibrin in blood clots. Besides fibrinolysis, plasmin is also involved in such physiological and pathological processes as wound healing, liver repair, and the maintenance of liver homeostasis. The SensoLyte® AFC Plasmin Activity Assay Kit provides a convenient assay for high throughput screening of plasmin inhibitors and inducers or for continuous assay of plasmin activity. The assay utilizes a fluorogenic peptide, which upon enzyme cleavage releases the AFC (7-amido-4-trifluoromethylcoumarin) fluorophore. The fluorescent signal can be measured at Ex/ Em =380 nm/500 nm. This assay can detect picogram level of plasmin activity. Assays are performed in a convenient 96-well microplate format. Kit size: 100 assays | |
Detailed Information | ![]() ![]() |
Product Citations | Ambadapadi, S. et al. (2015) Reactive Center Loop (RCL) peptides derived from serpins display independent coagulation and immune modulating activities J. Biol. Chem. 291 2874-2887doi: 10.1074/jbc.M115.704841Portelli, MA. et al. (2014). Genome-wide protein QTL mapping identifies human plasma kallikrein as a post-translational regulator of serum uPAR levels. FASEB J 28, 923.Park, HJ. et al. (2013). Identification of a specific haptoglobin C‐terminal fragment in arthritic synovial fluid and its effect on interleukin‐6 expression. Immunol 140, 133. doi: 10.1111/imm.12125.Sasaki, T. et al. (2012). Intravenous and oral administrations of DD2 [7-Amino-2-(sulfanylmethyl)heptanoic acid] produce thrombolysis through inhibition of plasma TAFIa in rats with tissue factor-induced microthrombosis. Thrombosis Res 130, e222. doi:10.1038/jid.2012.204Stewart, CE. et al. (2012). uPAR regulates bronchial epithelial repair in vitro and is elevated in asthmatic epithelium. Thorax 67, 477.Tudpor, K. et al. (2012). Urinary plasmin inhibits TRPV5 in nephrotic-range proteinuria. J Am Soc Nephrol 23, 1824. |
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PCR(聚合酶链式反应)是利用DNA在体外摄氏95°高温时变性会变成单链,低温(经常是60°C左右)时引物与单链按碱基互补配对的原则结合,再调温度至DNA聚合酶最适反应温度(72°C左右),DNA聚合酶沿着磷酸到五碳糖(5'-3')的方向合成互补链。
恒温PCR和实时荧光定量PCR的不同,是不同在实时荧光定量PCR的系统中加入了荧光染料(SYBR Green 或Taqman 探针等等)。以SYBR Green为例,这种染料可以结合在双链的DNA上,当PCR不断进行时,每一次退火生成的双链DNA也在增加,荧光染料结合也越多,荧光也越强。在机器中有一个探测荧光的探头,可以定量检测荧光的强度,转换成数值。这样就可以实时记录反映体系中DNA的反应情况。
荧光PCR更有优势,因为荧光PCR灵敏度高于恒温PCR,同样价格也高一些。
请教园友:
一般做mRNA表达的时候,需要注意提取的RNA中是否有DNA污染,或者通过设计跨内含子的引物来解决。那么,检测MmiRNA的时候,需要注意RNA中DNA污染的问题么?
荧光定量PCR较普通PCR不同的一点就是可以实时检测PCR扩增产物,从而可进行绝对定量或者相对定量。
荧光定量PCR较普通PCR不同的一点就是可以实时检测PCR扩增产物,从而可进行绝对定量或者相对定量。
一般都是相对量。则用delta delta CT方法来计算。举例如下:
对照组基因A的CT值为20, 内参(比如βactin)CT值15。实验组基因A CT值18,内参CT值14。
首先算加样量:delta CT=15-14=1。2的1次方是2。也就是说实验组的加样量是对照组的2倍。
基因A: delta CT=20-18=2。2的2次方是4。也就是说基因A的量在实验组是对照组的4倍。但是由于加样量是2倍,所以4处以2=2,最后的相对量是2倍。
几点注意:
1。必须确定扩增的特异性
2。 只有相同目标的CT值才能相减(扩增效率有可能不同)
3。 2的某次方只是理论值,实际扩增效率低于2。
4。 最好不用Syber Green
C代表的是cycle,循环数目,T代表的是threshold,阈值。
所以表达量越少的话,需要越多的循环才能扩增出来。也就是CT值越高

