| Clone | Polyclonal |
| Units | 1 ml |
| Host | Goat |
BackgroundThe reactivity of the antiserum is restricted to human alpha-2 macroglobulin. In immunoelectrophoresis and radial immunodiffusion, using various antiserum concentrations against normal human plasma a single precipitin line is obtained which shows a reaction of identity with the precipitin line obtained with purified alpha-2 macroglobulin. No reaction is obtained with any other plasma protein components or serum. In precipitating techniques as immunoelectrophoresis and single or double radial immunodiffusion (Mancini, Ouchterlony) to identify the presence of alpha-2 macroglobulin in human plasma or other body fluids or to determine its concentration. This antiserum is not intended for use in non-precipitating antibody binding or other highly sensitive assays, but this does exclude such use if proper controls are included.
SourceAlpha-2 macroglobulin(clotting factor I) is the major protein in the alpha-2 region of human serum after separation by immunoelectrophoresis. The concentration in normal plasma is about 2.5 mg/ml. It is a large polymer and a strong antigen. It forms about one third in the total of the alpha-2 globulins. The protein is isolated by ammonium sulphate precipitation, ion exchange chromatography, gelfiltration and preparative starch block electrophoresis. In the purification the activation of kallikrein is avoided, since it forms an irreversible bond with alpha-2 macroglobulin, thus contaminating the final immunogen. Freund’s complete adjuvant is used in the first step of the immunization procedure.
ProductDelipidated, heat inactivated, lyophilized, stable whole antiserum No preservative added as it may interfere with the antibody activity. Total protein and IgG concentrations in the antiserum are comparable to those of pooled normal goat serum.
Formulation: Delipidated, heat inactivated, lyophilized, stable whole antiserum No preservative added as it may interfere with the antibody activity. Total protein and IgG concentrations in the antiserum are comparable to those of pooled normal goat serum.
SpecificityPrecipitating polyclonal Goat antiSerum to Human alpha-2 macroglobulin
Species Reactivity: The antiSerum does not cross-react with any other component of Human plasma. Inter-species cross-reactivity is a normal feature of antibodies to plasma proteins since they frequently share antigenic determinants. Cross-reactivity of this antiSerum has been tested in double radial immunodiffusion anti several ser of other species with the following results: Bovine + Cat + Monkey ++ Mouse - Dog ± Goat - Rabbit + Rat -
ApplicationsPrecipitation assays. The lyophilized antiserum is shipped at ambient temperature and may be stored at +4°C; prolonged storage at or below -20°C. Reconstitute the lyophilized antiserum by adding 1 ml sterile distilled water. Dilutions may be prepared by adding phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance of the antiserum. Diluted antiserum should be stored at +4°C, not refrozen, and preferably used the same day.
Storage
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 Exalpha Biologicals accepts no liability for any inaccuracies or omissions in this information.
Protein Reference(s)
Database Name: UniProt
Accession number: P01023
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没有marker,怎么知道你做的蛋白大小?
没有参照物,怎么知道你跑的快不快?
没有尺子,怎么知道你的size大小?
凭嘴说吗?
抗体的特异性鉴定 抗体的特异性是指与相应抗原或近似抗原物质的识别能力。抗体的特异性高,它的识别能力就强。衡量特异性通常以交叉反应率来表示。交叉反应率可用竞争抑制试验测定。以不同浓度抗原和近似抗原分别做竞争抑制曲线,计算各自的结合率,求出各自在 IC50时的浓度,并按下列公式计算交叉反应率。 如果所用抗原浓度IC50浓度为pg/管,而一些近似抗原物质的IC50浓度几乎是无穷大时,表示 这一抗血清与其他抗原物质的交叉反应率近似为 0,即该血清的特异性较好。
抗体的亲和力 是指抗体和抗原结合的牢固程度。亲和力的高低是由抗原分子的大小、抗体分子的结合位点与抗原决定簇之间立体构型的合适度决定的。有助于维持抗原抗体复合物稳定的分子间力有氢键、疏水键、侧链相反电荷基因的库仑力、范德华力和空间斥力。亲和力常以亲和常数K表示,K的单位是L/mol,通常K的范围在 108 ~1010 /mol,也有多达 1014 /mol。抗体亲和力的测定对抗体的筛选,确定抗体的用途,验证抗体的均一性等均有重要意义。向左转|向右转
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
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。
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.
1取组织
(1) 灌注固定:暴露心脏,将注射针头插入左心室,剪开右心耳,快速灌注PBS,待右心耳处流出澄清液体,换4%多聚甲醛灌注到大鼠四肢强直为止,快速取脑和肾。
(2) 固定:将组织置于4%多聚甲醛中固定15小时
(3) 脱水:将组织放入30% 蔗糖溶液中直至下沉
(4) 切片:脑组织冰冻切片20微米,37度烘干2h以上;
(5)将切片放置-80度保存,随用随取。
2免疫单标记
(1)做抗原修复(0.4g柠檬酸、3g柠檬酸钠、1000ml水配制成溶液,放到水浴箱中90~94℃保存15min),冷却后, 用1*PBS反复洗标本10min,吸去PBS,同时向标本加渗透液(配0.4%trition液,然后将BSA加入到配好的trition液中,配成1%浓度)100ul,常温下湿盒反应1h。
(2) 吸去渗透液,同时向标本加block液(5%封闭用正常山羊血清)100ul,常温下湿盒反应5h。
(3) 吸去block液,同时向标本加入1:50稀释的羊抗鼠一抗100ul,37度湿盒反应过夜。
(4)吸去一抗,并回收,用1*PBS洗液反复洗15min。
(5)快速从冰箱中取出驴抗羊的二抗,并立即用锡纸包裹,注入到一个新的以锡纸包裹的tube管中并用PBS以1:200稀释。
(6)进入暗室,吸去PBS,同时加入二抗100ul,37度湿盒反应2h,封片。
(7)放置到荧光显微镜下观察。

