Description
Alpha-1-Antitrypsin A1AT Paired Antibody Set
Affinity’s Alpha-1-Antitrypsin A1AT Paired Antibody Set consists of matched capture and detecting antibodies that have been titrated and optimized for use in sandwich style ELISA assays. The product as provided contains sufficient capture and detecting antibodies for five full 96-well microplates and contains a detailed protocol sheet containing directions for use, recipes for solutions and sources for additional materials required. This Alpha-1-Antitrypsin A1AT Paired Antibody Set is intended to facilitate the end user in establishing an “in-house” immunoassay for research purposes only and must not be used for diagnostic applications. Assay validation is the responsibility of the end user.
Product Code: A1AT-EIA
Supplied Materials:
- Capture Antibody (A1AT-EIA-C): One yellow-capped vial containing 0.5 ml of polyclonal affinity purified anti-A1AT antibody for coating plates.
- Detecting Antibody (A1AT-EIA-D): One red-capped tube containing 0.5 ml of peroxidase conjugated affinity-purified polyclonal anti-A1AT antibody for detection of captured Alpha-1-Antitrypsin.
Related Products: VisuLize Buffer Pak
Species Cross Reactivity: View Chart
Product Datasheet: Alpha-1-Antitrypsin A1AT Matched Pair Antibody Set for ELISA
Description of Alpha 1-Antitrypsin A1AT
Alpha-1-Antitrypsin A1AT, also known as Alpha-1-Proteinase inhibitor (A1PI), is the most abundant protease inhibitor in blood and a member of the SERPIN family of proteinase inhibitors. Serum levels are typically 1.3 mg/ml (25 μM) but A1AT is an acute phase protein and concentrations can rise four-fold during inflammatory episodes or tissue injury. Low levels in circulation have been associated with pulmonary disease such as emphysema. A1AT is a single chain molecule with a mass of 52,000 daltons that is produced primarily in the liver and to a lesser extent by blood monocytes and intestinal epithelium. Based on association rates, the primary target enzyme for A1AT is believed to be neutrophil elastase1,2, but A1AT is a broad-spectrum inhibitor for many serine proteinases and the main role of A1AT in vivo is likely that of a “backup” inhibitor and proteinase scavenger in fluids and tissues. Although the association rates of A1AT with other enzymes are lower, the high concentration in plasma makes it an important inhibitor of activated Protein C, activated FXI, thrombin and plasmin1-4. Enzyme inhibition by A1AT occurs through proteolytic cleavage between Met358 and Ser359, which induces a conformational change in A1AT locking the enzyme into a stable, inactive 1:1 enzyme-inhibitor complex.
References and Reviews
- Johnson D, Travis J; Oxidative Inactivation of Human α-1-Proteinase Inhibitor; JBC 254, pp4022-4026, 1979.
- Travis J, Johnson D; Human α-1-Proteinase Inhibitor; Methods in Enzymology, 80, pp 754-765, 1981.
- Heeb MJ, Griffin JH; Physiologic Inhibition of Human Activated Protein C by α-1-Trypsin Inhibitor; JBC 263, pp11613-11616, 1988.
- Scott CF, Schapira M, James HL, Cohen AB, Colman RW; The Inactivation of Factor XIa by plasma protease inhibitors: Predominant role of α1protease inhibitor and protective effect of high molecular weight kininogen. J Clin Invest 69, pp 844, 1982.
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又由于自然存在的抗原大都存在多个抗原表位,会刺激机体产生多种针对同一抗原的不同抗原表位相应的不同抗体.
单克隆抗体这项新技术从根本上解决了在抗体制备中长期存在的特异性和可重复性问题,可用于探讨: ①蛋白质的精细结构;②淋巴细胞亚群的表面新抗原;③组织相容性抗原;④激素和药物的放射免疫(或酶免疫)分析;⑤肿瘤的定位和分类;⑥纯化微生物和寄生虫抗原;⑦免疫治疗和与药物结合的免疫-化学疗法 (“导弹”疗法,利用单克隆抗体与靶细胞特异性结合,将药物带至病灶部位.。
因此,单克隆抗体可直接用于人类疾病的诊断、预防、治疗以及免疫机制的研究,为人类恶性肿瘤的免疫诊断与免疫治疗开辟了广阔前景。
1.单克隆抗体的优点:
(1)杂交瘤可以在体外“永久”地存活并传代,只要不发生细胞株的基因突变,就可以不断的生产高特异性、高均一性的抗体.
(2)可以用相对不纯的抗原,获得大量高度特异的、均一的抗体.
(3)由于可能得到“无限量”的均一性抗体,所以适用于以标记抗体为特点的免疫学分析方法,如IRMA和ELISA等.
(4)由于单克隆抗体的高特异性和单一生物学功能,可用于体内的放射免疫显像和免疫导向治疗.
2.单克隆抗体的局限性:
(1)单克隆抗体固定的亲和性和局限的生物活性限制了它的应用范围.由于单克隆抗体不能进行沉淀和凝集反应,所以很多检测方法不能用单克隆抗体完成.
(2)单克隆抗体的反应强度不如多克隆抗体.
(3)制备技术复杂,而且费时费工,所以单克隆抗体的价格也较高.
说得简单点就是,细胞核是原封不动的,没有重组过,而细胞核外的东西是很多别的细胞中提取出来的并组合起来的。这样的混合体淋巴细胞产生的抗体具有很强的嵌合能力,针对性比一般的抗体强,但是它的来源没变,所以称之为混合单克隆抗体
不过多克隆抗体因为有多个结合位点,事实上的特异性并不如单克隆抗体,所以一般Western blot都尽量要求使用单克隆抗体。以保证实验结果准确可靠