Description
PAI-1 – Plasminogen Activator Inhibitor type 1 Polyclonal Antibody – HRP Conjugated
Affinity’s PAI-1 – Plasminogen Activator Inhibitor type 1 Polyclonal Antibody – HRP Conjugated is the base level of our horseradish peroxidase conjugated PAI-1 – Plasminogen Activator Inhibitor type 1 antibodies. The purity of IgG is typically 90% and is provided in a solution of HEPES buffered saline containing 50% glycerol (v/v). The titre is essentially the same as the starting antiserum and each vial typically contains the amount of IgG recovered from one milliliter of antiserum however this IgG has been conjugated with Horseradish Peroxidase as an enzyme reporter. This PAI-1 – Plasminogen Activator Inhibitor type 1 Polyclonal Antibody – HRP Conjugated is generally intended for use as labeled primary antibodies in applications such as immunoassay and immunoblotting.
Product Code: SAPAI-HRP
Retail Product Size: 0.2mg vial
Host Animal: Sheep Anti-Human PAI-1 – Plasminogen Activator Inhibitor type 1 Polyclonal Antibody – HRP Conjugated
Species Cross Reactivity: View Chart
Product Datasheet: PAI Plasminogen Activator Inhibitor type-1 Polyclonal Antibody, hrp conjugated anti-human sheep IgG
Description of PAI-1 (Type-1 Plasminogen Activator Inhibitor)
Type I plasminogen activator inhibitor, PAI-1, is a 50 kDa single-chain glycoprotein which belongs to the serine protease inhibitor (SERPIN) family of proteins. The concentration of PAI-1 in normal human plasma is very low, with an average concentration of approximately 20 ng/ml. The plasma concentrations of PAI-1 can be affected by a number of factors including diurnal variations, age, sex, pregnancy, obesity and exercise status. PAI-1 is synthesized by various cell types including endothelial cells, hepatocytes, vascular smooth muscle cells, fibroblasts, mesothelial cells, granulosa cells and malignant cell lines. It is also found in the α-granules of platelets as well as plasma. PAI-1 exists in at least three different conformations, including an active form with a half-life of approximately 1 – 3 hours, a latent form and a proteolytically or oxidatively inactivated form. The plasma glycoprotein vitronectin has been shown to bind active PAI-1 and stabilize it in its active conformation, extending its functional half-life to greater than 24 hours. Little specific binding of the latent or inactive forms of PAI-1 to vitronectin occurs. PAI-1 is considered to be the primary regulator of plasminogen activation in vivo. It is the major physiologic inhibitor of both the single chain and two-chain forms of tPA, being able to inhibit the single-chain form at least 10000 times greater than other PAIs.1-3
References and Reviews
- Kruithof, E.K.O. Plasminogen activator inhibitor type 1: biochemical, biological and clinical aspects. Fibrinolysis, 2:59-70, 1988.
- Edelberg, J.M, Sane, D.C., and Pizzo, S.V. Vascular regulation of plasminogen activator inhibitor-1 activity. Semin. Thromb. Hemost., 20(4):319-323, 1994.
- Loskutoff, D.J., Sawdey, M., and Mimuro, J. Type 1 plasminogen activator inhibitor, Prog. Thromb. Haemost., 9:87-115, 1988.
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1、直接法:这是最早的方法。其基本原理是用已知的抗体标记上荧光素后成为特异性荧光抗体,染色时将该抗体直接滴在载玻片上进行孵育,使之直接与载玻片上的抗原结合,在荧光显微镜下直接观察,作出判断。该方法评价:简单易行、特异性高、快速方便,常用于肾活检组织几种免疫球蛋白的检测和病原体的检测。但其不足是只能检测相应的一种物质,敏感性较差,效果有时不理想,目前较少作为更多方面的检测。
2、间接法:该法的基本原理是用特异性的抗体与切片中的抗原结合后,继用间接荧光抗体,与前面的抗原抗体复合物结合,形成抗原抗体荧光复合物。在荧光显微镜下,根据复合物的发光情况来确定所检测的抗原。该方法评价:由于结合在抗原抗体复合物上的荧光素抗体增多,发出的荧光亮度强,因而其敏感性强。目前本法应用较广泛,只需制备一种种属荧光抗体,即可适用于多种第一抗体的标记显示。
3、如果你有针对待查抗原一抗(荧光素标记),而且你的待检抗原表达也比较丰富的话,做直接法也未尝不可。不过如果你不具备上述两个条件的话,还是推荐你做间接法。
4、如果你做的是细胞骨架,那么就是用直接免疫荧光。如果是一般的蛋白表达,这个要看有没有试剂是直接标记的;如果没有,还是只能考虑间接荧光。直接荧光相对间接荧光可以避免非特异姓染色,当然步骤也少了些。我做过细胞骨架的直接染色,效果很好。
此法适用于小量抗体的荧光素标记,标记简便,非特异性染色较少。
(1)试剂和材料 试剂和材料同改良法。
(2)方法及步骤
①用0.025mol/L碳酸盐缓冲液pH9.0,将欲标记免疫球蛋白稀释成1%浓度,装入透析袋中。
②用同一缓冲液将FITC配成0.1mg/ml的溶液,按10mg/ml球蛋白溶液体积的10倍,将FITC稀释液盛于圆柱形容器内,并使透析袋浸没于FITC液中。
③容器顶端盖紧,底部放搅拌棒,在4~C电磁搅拌下透析标记24h。取出透析袋中标记液,即刻用SephadexG50凝胶过滤,去除游离荧光素,分装,贮存于4℃中。
2.放射免疫检测放射免疫检测技术是目前灵敏度最高的检测技术,利用放射性同素标记抗原(或抗体),与相应抗体(或抗原)结合后,通过测定抗原抗体结合物的放射性检测结果.放射性同位素具有pg级的灵敏度,且利用反复曝光的方法可对痕量物质进行定量检测.但放射性同位素对人体的损伤也限制了该方法的使用.
3.酶联免疫吸附试验(ELISA)酶联免疫检测是目前应用最广泛的免疫检测方法.该方法是将二抗标记上酶,抗原抗体反应的特异性与酶催化底物的作用结合起来,根据酶作用底物后的显色颜色变化来判断试验结果,其敏感度可达ng水平.常见用于标记的酶有辣根过氧化物酶(HRP)、碱性磷酸酶(AP)等.由于酶联免疫法无需特殊的仪器,检测简单,因此被广泛应用于疾病检测.常用的方法有间接法、夹心法以及BAS-ELISA.间接法是先将待测的蛋白抱被在孔板内,然后依次加入一抗、标记了酶的二抗和底物显色,通过仪器(例如酶标仪)定量检测抗原.这种方法操作简单但由于高背景而特异性较差.目前已逐渐被夹心法取代.夹心法利用二种一抗对目标抗原进行捕获和固定,在确保灵敏度的同时大大提高了反应的特异性.近年来,抗原的定量检测技术也不断推陈出新.近年来,在夹心法ELISA的基础上,开发了多抗原检测试剂盒,能同时检测微量液相样本中多个抗原含量.这项技术的应用大大缩短了诊断的时间,提高诊断的可靠性和及时性.
4.免疫金胶体技术胶体金技术经过30多年的发展到现在已日趋成熟,该方法是将二抗标记上胶体金颗粒,利用抗原抗体间的特异性反应,最终将胶体金标记的二抗吸附于渗滤膜上,此方法简单,快速,广泛应用于临床筛查.
SPSS软件统计结管归析其析都看SIGSIG=significance意显著性面值统计P值P值0.01<P<0.05,则差异显著P<0.01,则差异极显著
F值差检验量整模型整体检验看拟合程没意义
t值每自变量(logistic归)逐检验看beta值β即归系数没意义
T数值表示归参数显著性检验值绝值于等于ta/2(n-k)(值表示根据置信水平自由度数值)拒绝原假设即认其解释变量变情况解释变量X解释变量Y影响显著
F值归程显著性检验表示模型解释变量与所解释变量间线性关系总体否显著做推断若F>Fa(k-1,n-k),则拒绝原假设即认列入模型各解释变量联合起解释变量显著影响反则显著影响
二抗可以耦联有几种不同的标记,可以是酶,荧光素,或生物素。