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- Peptide (C)GRVRTYQFDSFLESTR, corresponding to amino acid residues 97-112 of mouse BAI1 (Accession Q3UHD1). Extracellular, N-terminus.
- 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.
- 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).
- 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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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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