Description | alpha-tubulin (b-alpha-1) mRNA is expressed only in brain with a molecular weight of about 55,000. The 3-prime UTR of b-alpha-1 is more than 80% homologous to the UTR of the rat brain alpha-tubulin gene, IL-alpha-T1. B-alpha-1 encodes a predicted 451-amino acid protein that is 100% identical to the rat homolog and differs by only 2 and 3 amino acids from the pig and chicken homologs, respectively. |
Catalog Number | IW-MA1107 |
Quantity | 9 ml |
Host | Mouse |
Clone | Tub-1 |
Isotype | Mouse IgG1 |
Immunogen | Microtubules from chicken embryo brain. |
Purity | Purified by the goat anti-mouse IgG affinity chromatography. |
Conjugate | Unconjugated |
Species Reactivity | Human, rat. Not tested in other species. |
Positive Control | Brain |
Cellular Localization | Cytoplasmic |
Form | Ready to use solution in PBS with stabilizer and 0.01% sodium azide. No further dilution needed. Serum blocking step should be omitted. |
Storage | Store at 2-8 °C. Do not freeze. |
Applications | IHC-P: Heat induced epitope retrieval is required on formalin fixed paraffin sections. IHC-Fr: Not tested. ICC: Not tested. |
Limitations | This product is intended for Research Use Only. Interpretation of the test results is solely the responsibility of the user. |
Precautions | Users should follow general laboratory precautions when handling this product. Wear personal protective equipment to avoid contact with skin and eyes. |
References | 1. Cowan, N. J.; Dobner, P. R.; Fuchs, E. V.; Cleveland, D. W. : Expression of human alpha-tubulin genes: interspecies conservation of 3-prime untranslated regions. Molec. Cell. Biol. 3: 1738-1745, 1983. 2. Hall, J. L.; Cowan, N. J. : Structural features and restricted expression of a human alpha-tubulin gene. Nucleic Acids Res. 13: 207-223, 1985. |
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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.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.
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。