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Antigen Incl. This antibody is shipped with its antigen FREE of charge!
- Peptide (C)GRVRTYQFDSFLESTR, corresponding to amino acid residues 97-112 of mouse BAI1 (Accession Q3UHD1). Extracellular, N-terminus.

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Western blot analysis of rat (lanes 1 and 3) and mouse (lanes 2 and 4) brain lysates:1,2. Anti-BAI1 (extracellular) Antibody (#ABR-021), (1:200).
3,4. Anti-BAI1 (extracellular) Antibody, preincubated with the negative control antigen.
Western blot analysis of human HL-60 promyelocytic leukemia cell lysates:1. Anti-BAI1 (extracellular) Antibody (#ABR-021), (1:200).
2. Anti-BAI1 (extracellular) Antibody, preincubated with the negative control antigen.
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Expression of BAI1 in mouse olfactory bulbImmunohistochemical staining of mouse perfusion-fixed olfactory bulb frozen sections using Anti-BAI1 (extracellular) Antibody (#ABR-021), (1:200). A. BAI1 (green) is expressed in astrocyte-like cells (arrows). B. Double-staining of BAI1 (green) and glial fibrillary acidic protein (red) reveals expression of BAI1 in a subset of astrocytes. Nuclear staining of cells using the DNA dye DAPI (blue).
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Cell surface detection of BAI1 in live intact human HL-60 promyelocytic leukemia cell line:___ Unstained cells + goat-anti-rabbit-AlexaFluor-488 secondary antibody.
___ Cells + Anti-BAI1 (extracellular) Antibody (#ABR-021), (1:20) + goat-anti-rabbit-AlexaFluor-488 secondary antibody.The negative control antigen is not suitable for this application.
- 1. Park, D. et al. (2007) Nature 450, 430.
- 2. Cork, S.M. et al. (2011) J. Mol. Med. 89, 743.
- 3. de Fraipont, F. et al. (2001) Trends Mol. Med. 7, 401.
- 4. Oda, K. et al. (1999) Cytogenet. Cell. Genet. 84, 75.
- 5. Cork, S.M. et al. (2012) Oncogene 31, 5144.
- 6. Shiratsuchi, T. et al. (1997) Cytogenet. Cell. Genet. 79, 103.
- 7. Hatanaka, H. et al. (2000) Int. J. Mol. Med. 5, 181.
- 8. Fukushima, Y. et al. (1998) Int. J. Oncol. 13, 967.
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The three members of the brain angiogenesis inhibitor (BaI1-3) are receptors belonging to the adhesion subfamily of G-protein coupled receptor superfamily. Like all members of GPCRs, all three BaIs have seven transmembrane domains, an intracellular C-terminal tail and extracellular N-terminus. Like other adhesion members, the N-terminus is quite large1,2. Many domains are localized to the N-terminus; various glycosylations sites are present, there is a GPCR proteolysis site, a putative hormone binding domain and thrombospondin type 1 repeats which regulate the anti-angiogenic activity of thrombospondin-12,3. The C-terminal tail interacts with PDZ-domain proteins. Unique to BaI1 is a proline-rich domain required for interacting with Src homology domains and WW domain proteins2,4.
Like most adhesion GPCRs, BaI also undergo proteolysis at the N-terminus at a highly rich cystein domain2. Following autocleavage, the N-terminal fragment remains associated to the receptor. In BaI1, proteolysis yields a partly secreted 120 kDa. fragment (vasculostatin-120) or a 40 kDa. fragment both having antiangiogenic effects2,5.
At the mRNA level, all BaIs are expressed in fetal and adult human brain2,6. BaI2 is detected in the human heart and skeletal muscle. BaI3 is expressed in the human heart, testis and small intestine. In mouse, both BaI2 and BaI3 are restricted to the brain2.
These receptors are implicated in various diseases and disorders such as primary glioma, pulmonary adenocarcinomas, gastric and colorectal cancers2,6,7.
Anti-BAI1 (extracellular) Antibody (#ABR-021) is a highly specific antibody directed against an epitope of the mouse protein. The antibody can be used in western blot, immunohistochemistry, and indirect live cell flow cytometry applications. It has been designed to recognize BAI1 from rat, mouse, and human samples.
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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、制备抗原。
2、选择实验动物。
3、动物免疫。
4、试取血进行测试,看看是否成功免疫。
5、如果成功免疫,杀死实验动物,采集全部血清。
6、纯化出抗体。
7、鉴定抗体。包括纯度以及特异性。
二、抗体分类
抗原通常是由多个抗原决定簇组成的,由一种抗原决定簇刺激机体,由一个B淋巴细胞接受该抗原所产生的抗体称之为单克隆抗体(Monoclone antibody)。由多种抗原决定簇刺激机体,相应地就产生各种各样的单克隆抗体,这些单克隆抗体混杂在一起就是多克隆抗体,机体内所产生的抗体就是多克隆抗体;除了抗原决定簇的多样性以外,同样一类抗原决定簇,也可刺激机体产生IgG、IgM、IgA、IgE和IgD等五类抗体。多克隆抗体是由异源抗原(大分子抗原、半抗原偶联物)刺激机体产生免疫反应,有机体浆细胞分泌的一组免疫球蛋白。多克隆抗体由于其可识别多个抗原表位、可引起沉淀反应,制备时间短,成本低的原因广泛应用于研究和诊断方面。
三、免疫方法
可以采用以下各种方法之一进行免疫。
(1)淋巴结注射法:①在兔的两后足跖部皮下(或皮内)注射活卡介苗 50mg(每侧约0.30ml) 。7~10 天后,兔跖及腘肌淋巴结肿大;②于肿大的两侧淋巴结内各注射加有完全佐剂的IgG 乳化抗原 0.50ml(含 IgG 5mg/ml、青霉素 1 000U/ml、链霉素 1 000μg/ml);③必要时,14 天后,重复步骤②一次;④再过 7 天后,于两侧淋巴结内各注射加有完全佐剂的 IgG 乳化抗原0.50ml(含IgG5mg/ml、青霉素 1 000U/ml、链霉素 1 000μg/ml) ;⑤5~7天后,耳静脉采血。测定血清效价。
(2)皮下多点注射法:①家兔两侧掌(跖内各注射含有完全佐剂抗原 0.10ml(IgG 含量5mg/ml) ;②7~10 天后,脊柱两侧多点(颈、胸、腰椎各两点、共 6 点)皮下注射含不完全佐剂5的抗原,每点 0.50ml;③7~10 天后,脊柱两侧重复注射一次;④7~10 天后试血。不合格者重复步骤③。
(3)多途径联合注射法:①两侧掌(跖)内侧皮下注射含完全佐剂抗原 0.50ml(IgG 量为 5mg/ml) ;②14 天后,多点皮下注射含有不完全佐剂抗原;③7 天后,耳静脉注射不含佐剂的抗原 2ml;④测定血清抗体效价,不合格者重复步骤③,并适当递增 IgG 量。
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.
单克隆抗体结构相似且均一,有高度特异性
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。
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。

