抗体来源 Rabbit
克隆类型 polyclonal
交叉反应 Human, Mouse, Rat, Dog, Cow, Horse, Rabbit
产品类型 一抗
研究领域 肿瘤 细胞生物 免疫学 细胞周期蛋白
蛋白分子量 predicted molecular weight: 122kDa
性 状 Lyophilized or Liquid
免 疫 原 KLH conjugated synthetic peptide derived from human Bub1
亚 型 IgG
纯化方法 affinity purified by Protein A
储 存 液 0.01M PBS, pH 7.4 with 10 mg/ml BSA and 0.1% Sodium azide
产品应用 WB=1:100-500 ELISA=1:500-1000 IP=1:20-100 IHC-P=1:100-500 IHC-F=1:100-500 IF=1:100-500
（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.
纺锤体检查点蛋白抗体保存条件 Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.
Important Note This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
产品介绍 Bub1 was initially identified as the mammalian homolog to the yeast protein through a cDNA screen. Bub1 encodes a kinase involved in the formation of the spindle checkpoint, an important mechanism that ensures high fidelity mitotic chromosome segregation. It is thought to be required for assembly of a functional inner centromere, sister chromatid cohesion via targeting of the Shugoshim protein and metaphase congression. Bub1 functions by phosphorylating cdc20, a member of the mitotic checkpoint complex and activating the spindle checkpoint. A related protein kinase Bub3 interacts with Bub1 and targets it to kinetochores prior to chromosome alignment. Mutations in bub1 have been associated with aneuploidy and several forms of cancer.
Function : Serine/threonine-protein kinase that performs 2 crucial functions during mitosis: it is essential for spindle-assembly checkpoint signaling and for correct chromosome alignment. Has a key role in the assembly of checkpoint proteins at the kinetochore, being required for the subsequent localization of CENPF, BUB1B, CENPE and MAD2L1. Required for the kinetochore localization of PLK1. Plays an important role in defining SGOL1 localization and thereby affects sister chromatid cohesion. Acts as a substrate for anaphase-promoting complex or cyclosome (APC/C) in complex with its activator CDH1 (APC/C-Cdh1). Necessary for ensuring proper chromosome segregation and binding to BUB3 is essential for this function. Can regulate chromosome segregation in a kinetochore-independent manner. Can phosphorylate BUB3. The BUB1-BUB3 complex plays a role in the inhibition of APC/C when spindle-assembly checkpoint is activated and inhibits the ubiquitin ligase activity of APC/C by phosphorylating its activator CDC20. This complex can also phosphorylate MAD1L1. Kinase activity is essential for inhibition of APC/CCDC20 and for chromosome alignment but does not play a major role in the spindle-assembly checkpoint activity. Mediates cell death in response to chromosome missegregation and acts to suppress spontaneous tumorigenesis.
Subunit : Interacts with BUB3 and CASC5. Interacts (when phosphorylated) with PLK1. The BUB1-BUB3 complex interacts with MAD1L1. Interacts with SV40 Large T antigen; this interaction induces activation of a DNA damage response and promotes p53/TP53 stabilization and phosphorylation (Probable).
Subcellular Location : Nucleus. Chromosome, centromere, kinetochore.
Tissue Specificity : Highly expressed in thymus followed by spleen. Preferentially expressed in tissues with a high mitotic index.
Post-translational modifications : Phosphorylated upon DNA damage, probably by ATM or ATR. Upon spindle-assembly checkpoint activation it is hyperphosphorylated and its kinase activity toward CDC20 is stimulated. Phosphorylation at Thr-609 is required for interaction with PLK1, phosphorylation at this site probably creates a binding site for the POLO-box domain of PLK1, thus enhancing the PLK1-BUB1 interaction.
Ubiquitinated and degraded during mitotic exit by APC/C-Cdh1.
Similarity : Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. BUB1 subfamily.
Contains 1 BUB1 N-terminal domain.
Contains 1 protein kinase domain.
Database links : UniProtKB/Swiss-Prot: O43683.1
纺锤体检查点（spindle checkpoint)是细胞周期中的一个重要检查点，由Mad2、Bub1等染色体动粒(kinetochore)蛋白构成，像“传感器”一样能感知微管与动粒结合情况和张力。纺锤体检查点阻滞细胞进入分裂后期直到所有染色体着丝点正确与微管联接,确保染色体均等分配,维持基因组稳定性。
纯度：在实验的任何阶段，确定抗体溶液纯度的最简单方法是取一部分样本进行SDS-PAGE电泳。凝胶可用考马斯亮蓝染色(灵敏度为0.1—0.5ug/带)或银染(灵敏度1～l0ug/带)。
定量：如果抗体还不纯，有一个快捷的定量方法，即通过SDS-PAGE电泳分离出轻、重链，然后和已知的标准染色带比较。如果需要分析许多样本，用免疫测定法对抗体定量较容易。如果抗体是经过纯化的，可通过测蛋白总量代替上述两种方法，有一简单的方法，即紫外吸收法。纺锤体检查点蛋白抗体的量可通过测280nm处的吸收值来测(10D大致相当于0.75mg/m1的纯化抗体)。
抗原结合活性：一般说来，纯化方法不会引起抗原结合活性的改变。用蛋白G或蛋白A树脂很少导致抗体活性丧失。然而，如果最终抗体产物的作用不如原来所预料的好，检测抗体纯化过程所丢失的活性就极为重要。用一系列滴定法比较纯化的抗体和其原材料的活性，以标定每一步中的总抗体量，这将有助于较好的估计通过纯化所丢失的活性。
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