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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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多抗,稀释度更大,特异性相对较差,容易出现多条带。
兔的单克隆抗体和鼠的单克隆抗体在使用上不会有什么区别。
用来很多抗体,许多时候觉得单抗多抗也未必是理论上那样的。单抗做不好的也有,多抗条带唯一且清晰的也有。
而且很多蛋白的抗体未必有那么多的选择。
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。
单克隆抗体结构相似且均一,有高度特异性
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.
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。