- Overview
- Data/Specifications
- Literature/Support
- How It Works
- Related Products
Overview
Human HtrA1 protease was initially identified in human fibroblasts and belongs to the high temperature requirement factor A (HtrA) family of serine proteases that can be distinguished from other serine proteases by sequence homology, by the presence of a trypsin-type protease domain and one or two carboxyterminal PDZ domains and by their oligomeric architecture.
The loss of mammalian HtrA activity is correlated with severe diseases, including arthritis, cancer, familial ischemic cerebral small vessel disease and age related macular degeneration, as well as Parkinson’s disease and Alzheimer’s disease.
Epigenetic silencing occurs in various cancers, and the loss of HtrA1 correlates with decreased sensitivity to anticancer drugs and increased cell migration. In addition, overexpression of HtrA1 inhibited proliferation in vitro and tumor growth in vivo. These data suggest that HtrA1 might function as a tumor suppressor.
In the extracellular matrix, HtrA1 cleaves numerous secreted proteins, such as fibronectin, decorin, fibro- modulin, aggrecan, type II collagen, biglycan, clusterin, a disintegrin and metalloproteinase domain-contain- ing 9 (ADAM9), vitronectin, α-2-macroglobulin and the amyloid precursor protein fragment Aβ. The degra- dation of extracellular matrix components and the strong upregulation of HtrA1 in samples from patients implicate HtrA1 in arthritic diseases, in which it might affect the degradation of cartilage as well as inflam- mation.
It is unknown, how the cellular distribution of HtrA1 is regulated. In addition, little is known about the inter- action partners of mammalian HtrAs. It will be important to identify proteins that function as determinants of the cellular localization, substrate specificity and regulation of HtrA proteases.
Data/Specifications
Species: human
Sample Type: serum, tissue, cell culture supernates
Sample Size: 500 uL
Standard Curve Range: 0.391 - 25 ng/mL
Sensitivity: 391 pg/ml
Assay Length:4 hours
Literature/Support
Product Insert:
human HtrA1 ELISA Insert (PDF)
Articles/Troublshooting:
ELISA Troubleshooting Guide
ELISA Data Reduction Guide
How It Works
This quantitative assay is based on a two site sandwich format. A highly specific monoclonal antibody against HtrA1 is immobilised on the plate. HtrA1 will be bound to the wells, other components of the sample are removed by discarding/drying by taping and washing of the plate. The analyte is detected in two steps using a secondary biotin-labeled monoclonal antibody and a highly polymerised streptavidin-peroxidase conjugate. Any excess is removed by discarding/drying by taping and washing after each detection step. The amount of peroxidase bound to each well is determined by the addition of TMB Substrate. The reaction is stopped by adding the Stop Solution and the resultant color is read in a microplate reader at 450 nm. The concentration of HtrA1 in a sample is determined by interpolation from the standard curve.
Related Products
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MPI是一种基于消息传递的并行编程技术，在不同节点计算机之间并行多进程执行程序（这种情况下，不同享内存），只是通过消息传递来进行通信，从而适用于分布式体系
谢谢！！！
测器和微处理器控制系统等组成.
光源灯发出的光线经过滤光片或单色器后,成为一束单色光.该单色光束经过酶标板中的待测标本,被标本吸收掉一部分后,到达光电检测器.光电检测器将投照到
上面的光信号的强弱转变成电信号的大小.此电信号经前置放大、对数放大、模数转换等处理后,送人微处理器进行数据处理和计算,最后通过显示器和打印机输出
测试结果.
3.1透过率、吸收度测量
a.暗电流调零：开启电源后，若试样室盖未打开，将显示提示符P.1（请做第一步骤操作），打开试样室盖直到接触检测开关，提示符P.1消失，显示暗电流值。如果它的绝对值小于5.0，则两秒内自动归零，否则可调节面板上之0%T旋钮。
b.调整参比讯号（100T/OA）：微机所显示的提示符为P.2（请做第二步操作）。将T—A—C旋钮开关置T/A，参比溶液放进1号试样槽并移入光路，关闭试样室盖，显示参比讯号强度，调整狭缝和光楔（试样槽拉手左面之100%T旋钮），使显示数在90.0—119.9%T或相应A值范围以内，2秒以内将自动调整为100%T/OA。
c.比色皿误差校正：为校正比色皿的系统误差，要求在同一次测量过程中，比色皿与槽位编号一一照应。将使用的比色皿盛参比溶液放进试样槽，移入光路，所显示的T（或A值）就反映了该与1号槽中比色皿之间的配对误差。按一下ADJ（Adjust）校正键，则误差数据存入微机并自动校正为100%T/OA。这项操作在每次开机后只需作一次，若所用的比色皿配对性很好，可免去这一步骤。开机时，校正系数初始值为1，即相当于不校正。
d.用同比色皿盛被测溶液放进同一试样槽，则显示校正后的透过率或吸光率。
3.2浓度测量
a.标准对照法
b.按3.1节（1）—（3）操作。
c.将标准浓度的样品放如光路。
d.旋钮开关至C，分别按三个浓度置数键，使显示数与已知浓度值一致，浓度值仅取其三为有效数字，小数点位置自定义。
e.将被测式样移入光路，则显示样品的浓度（比色皿误差已校正）。
3.2标准曲线法（标准曲线为直线）
a.按3.1节（1）—（3）操作。
b.旋钮至A， 调整光楔使A值变化至标准曲线高端某一点的坐标A0。
c.旋钮至C，按浓度置数键显示数与对应于A0的浓度值C0一致。
d.重新调整对比讯号后按（1）之（4）操作。
3.3测试完毕，依次关闭主机电源开关，电气箱开关，拔下电源插头，盖上护罩。
3.4认真填写使用操作记录。
最近想单位准备购买一台双荧光素酶报告基因检测仪，不知哪家公司有售，价格多少？谢谢！
