抗体制备过程:
1.材料与试剂
a.提取的动物 Ig
b.弗氏佐剂和弗氏不佐剂
d.实验动物 兔
e.其它材料及试剂
2、选择实验活体。
3、进行动物免疫实验。
4、试取血样进行测试,查看免疫效果。
5、如果免疫成功,杀死实验活体,采集全部血清。
6、纯化出抗体。
7、鉴定抗体。胎牛血清(无菌采制)
公司产品仅用于科研专业供应的抗体,是用于化学反应、分析化验、研究实验、教学实验、化学配方使用的纯净化学品,价格实惠,多种规格供应,售后完善。
英文名称: RNA polymerase II
中文名称: RNA聚合酶II抗体
规格:50ul、100ul、200ul
别 名:POLR2A; POLR2; DNA directed RNA polymerase II A; DNA-directed RNA polymerase II largest subunit; DNA-directed RNA polymerase II subunit A; RPB1_HUMAN.
研究领域:染色质和核信号 转录调节因子 表观遗传学
抗体来源:Rabbit
克隆类型:Polyclonal
交叉反应:Mouse, Rat, (predicted: Human, Dog, Pig, Cow, Horse, )
产品应用:WB=1:500-2000 ELISA=1:5000-10000 IHC-P=1:100-500 IHC-F=1:100-500 IF=1:50-200 (石蜡切片需做抗原修复)
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
理论分子量:217kDa
细胞定位:细胞核
性 状:Liquid
浓 度:1mg/ml
免 疫 原:KLH conjugated synthetic peptide derived from human Pol II/RNA polymerase II: 101-200/1970
亚 型:IgG
纯化方法:affinity purified by Protein A
缓 冲 液:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
注意事项:This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
产品介绍:RNA polymerase II (Pol II) is an enzyme that is composed of twelve subunits and is responsible for the transcription of protein-coding genes. Transcription initiation requires Pol II-mediated recruitment of transcription machinery to a target promoter, thereby allowing transcription to begin. The largest subunit of Pol II (referred to as RPB1 or RPB205) is a 1,840 amino acid protein that contains one C2H2-type zinc finger and a C-terminal domain comprised of several heptapeptide repeats. Although Pol II function requires the cooperation of all twelve subunits, the largest subunit conveys Pol II catalytic activity and, together with the second largest subunit, forms the active center of the Pol II enzyme. Additionally, the large subunit participates in forming the DNA-binding domain of Pol II, a groove that is necessary for transcription of the DNA template. Without proper function of the large subunit, mRNA synthesis and subsequent transcription elongation cannot occur.
DNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. Largest and catalytic component of RNA polymerase II which synthesizes mRNA precursors and many functional non-coding RNAs. Forms the polymerase active center together with the second largest subunit. Pol II is the central component of the basal RNA polymerase II transcription machinery. It is composed of elements that move relative to each other. RPB1 is part of the core element with the central large cleft, the clamp element that moves to open and close the cleft and the jaws that are thought to grab the incoming DNA template. At the start of transcription, a single stranded DNA template strand of the promoter is positioned within the central active site cleft of Pol II. A bridging helix emanates from RPB1 and crosses the cleft near the catalytic site and is thought to promote translocation of Pol II by acting as a ratchet that moves the RNA-DNA hybrid through the active site by switching from straight to bent conformations at each step of nucleotide addition. During transcription elongation, Pol II moves on the template as the transcript elongates. Elongation is influenced by the phosphorylation status of the C-terminal domain (CTD) of Pol II largest subunit (RPB1), which serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination and mRNA processing. Acts as a RNA-dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, acting both as a replicate and transcriptase for the viral RNA circular genome.
抗体基本结构:
免疫荧光技术的实验步骤:
一、准备好试剂与仪器:
磷酸盐缓冲盐水、荧光标记的抗体溶液、缓冲甘油、搪瓷桶三只、有盖搪瓷盒一只、荧光显微镜、玻片架、滤纸、37℃温箱等。
二、实验步骤
1.滴加0.01mol/L,pH7.4的PBS于待检标本片上,十分钟后弃去,使标本保持一定湿度。
2.滴加适当稀释的荧光标记的抗体溶液,使其覆盖标本,置于有盖搪瓷盒内,保温一定时间以三十分钟为参考。
3.取出玻片,置玻片架上,先用0.01mol/L,pH7.4的PBS冲洗后,再按顺序过0.01mol/L,pH7.4的PBS三缸浸泡,每缸三到五分钟,并不停地摇晃振荡。
4.取出玻片,用滤纸吸去多余水分,但不使标本干燥,加一滴缓冲甘油,以盖玻片覆盖。
5.立即用荧光显微镜观察。观察标本的特异性荧光强度。
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抗体的定义:
抗体(antibody),(免疫球蛋白不仅仅只是抗体)是一种由浆细胞(效应B细胞)分泌,被免疫系统用来鉴别与中和外来物质如细菌、病毒等的大型Y形蛋白质,仅被发现存在于脊椎动物的血液等体液中,及其B细胞的细胞膜表面。抗体能识别特定外来物的一个特征,该外来目标被称为抗原。
动物抗体功能分类:
①猪抗体:猪瘟抗体,猪蓝耳抗体,猪圆环病毒抗体,猪伪狂犬抗体,猪细小病毒抗体,猪口蹄疫抗体,猪流感抗体等。
②禽抗体:小鹅瘟抗体,鸭肝抗体抗体,鸭浆膜炎抗体,抗体,新城疫抗体等
③牛抗体:牛口蹄疫抗体,奶牛乳房炎抗体,牛流行热抗体,牛病毒性腹泻抗体,牛出血性败血症抗体等
④羊抗体:羊痘抗体,羊口蹄疫抗体,羊小反刍兽疫抗体,羊快疫抗体,羊肠毒血症抗体,羊猝疽抗体,羊黑疫抗体等。
⑤犬抗体:犬狂犬病抗体、犬瘟热抗体、犬副流感抗体、犬腺病毒抗体与犬细小病毒病抗体,狐貉水貂的伪狂犬抗体、细小病毒抗体、乙脑抗体等。
抗体的结构:
抗体是具有4条多肽链的对称结构,其中2条较长、相对分子量较大的相同的重链(H链);2条较短、相对分子量较小的相同的轻链(L链)。链间由二硫键和非共价键联结形成一个由4条多肽链构成的单体分子。轻链有κ和λ两种,重链有μ、δ、γ、ε和α五种。 整个抗体分子可分为恒定区和可变区两部分。在给定的物种中,不同抗体分子的恒定区都具有相同的或几乎相同的氨基酸序列。可变区位于"Y"的两臂末端。在可变区内有一小部分氨基酸残基变化特别强烈,这些氨基酸的残基组成和排列顺序更易发生变异区域称高变区。高变区位于分子表面,最多由17个氨基酸残基构成,少则只有2 ~ 3个。高变区氨基酸序列决定了该抗体结合抗原抗原的特异性。一个抗体分子上的两个抗原结合部位是相同的,位于两臂末端称抗原结合片段(antigen-binding fragment, Fab)。"Y"的柄部称结晶片段(crystalline fragment,FC),糖结合在FC 上。
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