Product Type | Immunohistochemistry Kit |
Units | 60 Tests |
Application | Immunohistochemistry |
BackgroundCell death occurs by two major mechanisms, necrosis and apoptosis. Apoptosis is also known as programmed cell death or ankoikis (a form of apoptosis which is induced by anchorage-dependent cells detaching from the surrounding extracellular matrix). Apoptosis leads to the elimination of cells without releasing harmful substances into the surrounding area. Too little or too much apoptosis plays a role in a great many diseases. When apoptosis functions inappropriately, cells that should be eliminated survive and potentially become immortal, as in cancer or leukemia. When apoptosis works overly well, too many cells may ‘die’ and the result may be grave tissue damage. This is the case in stroke and neurodegenerative disorders such as Alzheimer, Huntington and Parkinson diseases. The term ‘apoptosis’ refers only to the structural changes a cell goes through during the process of programmed cell death and not to the process itself. Classical necrotic cell death occurs due to noxious injury or trauma to the cell while apoptosis is an energy dependent mechanism that takes place during normal cell development. While necrotic cell death results in cell lysis, cellular apoptosis is characterized morphologically by cell shrinkage, nuclear pyknosis, chromatin condensation, and blebbing of the plasma membrane. Apoptosis is the result of a cascade of molecular and biochemical events involving endogenous endonucleases that cleave DNA into the prototypical ‘ladder of DNA fragments’ that may be visualized in agarose gels. Observation of oligonucleosomal DNA fragments by DNA laddering has long been the most acceptable and only available assay for the detection of apoptosis. Exalpha’s DNA Fragmentation Detection Kit exploits the fact that apoptotic endonucleases not only affect cellular DNA by producing the classical DNA ladder but also generate free 3’-OH groups at the ends of these DNA fragments. These free 3’-OH groups are end-labeled by the DNA Fragmentation Detection Kit allowing for the detection of apoptotic cells using a molecular biology-based, end labeling technique.
ApplicationsOptimal concentration should be evaluated by serial dilutions.
StorageExalpha’s DNA Fragmentation Detection Kit components are shipped on cold pack. Upon receipt, store kit at -20°C in a non-frost-free freezer. For long term storage, it is recommended that you aliquot and freeze the TdT Enzyme (Component 4), TdT Labeling Reaction Mix (Component 3), and 25x Streptavidin-HRP Conjugate (Component 7) at -20 °C. Thirty (30) minutes prior to use of each component, thaw component and keep on cold block or on ice. Return the components to -20°C for long term storage or 4-8ºC for short term storage (up to 2 weeks) immediately after use. Special care should be taken to keep TdT Enzyme (Component 4), TdT Labeling Reaction Mix (Component 3) and 25x Streptavidin-HRP Conjugate (Component 7) cold by pulling out the number of aliquots needed for the test, keeping them on ice, and leaving the remaining aliquots at -20°C.
CautionThis product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but Nordic-MUbio accepts no liability for any inaccuracies or omissions in this information.
Kit ManualClick here to download the Kit Manual Safety Datasheet(s) for this product:NM_X2044K SDS_V1/wp-content/uploads/SDS/X2044K SDS_V1.pdf
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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.
抗原有两个基本特性,即抗原性和免疫原性。有抗原性的物质不一定有免疫原性,所以由此引出半抗原和完全全抗原,半抗原必须经过经过一定的改造(偶联蛋白载体BSA,OVA或者HSA等大分子物质)方能成为完全。一般而言完全抗原分子量越大(大于10KDa),结构越复杂引起免疫反应的能力也就越强。
抗体就是能与特异性抗原结合的免疫球蛋白,抗体一般分为多克隆抗体和单克隆抗体,多克隆抗体能与抗原的多个表位结合。本篇主要讲述兔来源的多克隆抗体的生产步骤
多抗一般制备流程:完全抗原的准备→兔子的免疫→ 效价检测和终放→抗体亲和纯化→抗体的浓缩和保存。
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.
一般来说多克隆的阳性率高一些,但出现假阳性的比例也高一些。
其次,察看次目的蛋白的存在形式,有没有多聚体形式及变构形式;
最后,查看多家抗体公司的DATA,看看别人的WB做出来的条带的位置。
根据你说的,特异识别多个组织中的同样大小的条带,我觉得很可能就是你的目的蛋白。
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