抗体的生物素化标记实验要点：

1. 蛋白8抗体费用 如在反应混合液中有叠氮钠或游离氨基存在,会抑制标记反应。因此,蛋白质在反应前要对 0.1mol/L碳酸氢钠缓冲液或0.5mol/L硼酸缓冲液充分透析；

2.所用的NHSB及待生物素化蛋白质之间的分子比按蛋白质表面的ε-氨基的密度会有所不同,选择不当则影响标记的效率,应先用几个不同的分子比来筛选最适条件；

3.用NHSB量过量也是不利的,抗原的结合位点可能因此被封闭,导致抗体失活；

4.由于抗体的氨基不易接近可能造成生物素化不足,此时可加入去污剂如 Triton x-100, Tween20等；

5.当游离ε-氨基(赖氨酸残基的氨基)存在于抗体的抗原结合位点时,或位于酶的催化位点时,生物素化会降低或损伤抗体蛋白的结合力或活性；

6.生物素还可能与不同的功能基团,如羰基、氨基、巯基、异咪唑基及苯酚基,也可与糖基共价结合；

7.交联反应后,应充分透析,否则,残余的生物素会对生物素化抗体与亲和素的结合产生竞争作用；

8.在细胞的荧光标记实验中,中和亲和素的本底低,但由于链霉亲和素含有少量正电荷,故对某些细胞可导致高本底。

产品订购信息：
英文名称  Anti-Twinkle/ATXN8 

中文名称   蛋白8抗体费用 

别    名  Ataxin 8; Ataxin8; ATXN 8; ATXN8; C10 orf2; C10orf 2; C10orf2; Chromosome 10 open reading frame 2; IOSCA; PEO 1; PEO; PEO1; PEOA3; Progressive external ophthalmoplegia 1 protein; SANDO; SCA 8; SCA8; T7 gp4 like protein with intramitochondrial nucleoid localization; T7 like mitochondrial DNA helicase; Twinkle protein, mitochondrial; TWINL.

浓    度   1mg/1ml

规 格  0.2ml/200μg

抗体来源   Rabbit

克隆类型   polyclonal

交叉反应   Human, Mouse, Rat, Chicken, Dog, Pig, Cow, Horse, Rabbit 

产品类型   一抗  

研究领域    细胞生物 神经生物学 线粒体 表观遗传学

蛋白分子量  predicted molecular weight: 77kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from hu Twinkle/ATXN8 

亚    型   IgG

纯化方法   affinity purified by Protein A

储 存 液   Preservative: 15mM Sodium Azide, Constituents: 1% BSA, 0.01M PBS, pH 7.4

蛋白8抗体费用 产品应用    WB=1:100-500 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 ICC=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.  

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 Twinkle, also known as PEO1 (Progressive external ophthalmoplegia 1 protein), PEOA3, SANDO or TWINL, is a mitochondrial protein that functions as a 5’-3’ nucleotide-dependent DNA helicase. Colocalized with mtDNA (mitochondrial DNA) in mitochondrial nucleoids, Twinkle is important in the metabolism and maintenance of mtDNA, playing a crucial role in the regulation of mtDNA copy numbers. Twinkle is expressed at high levels in testis, pancreas and skeletal muscle and exists as three isoforms due to alternative splicing events. Defects in the gene encoding Twinkle are the cause of two conditions: progressive external ophthalmoplegia with mitochondrial DNA deletions autosomal dominant 3 (PEOA3) and sensory ataxic neuropathy dysarthria and ophthalmoparesis (SANDO). PEOA3 is characterized by ptosis and weak muscles, while SANDO is characterized by ophthalmoparesis, dysarthria and sensory ataxic neuropathies.

Function : Twinkle, also known as PEO1, is involved in mitochondrial DNA (mtDNA) metabolism, possibly by functioning as an adenine nucleotide-dependent DNA helicase at the mitochondrial DNA replication fork. It may be a key regulator of mitochondrial DNA copy number in mammals. Mutations in Twinkle lead to infantile-onset spinocerebellar ataxia, a severe neurodegenerative disorder. Mutations in Twinkle are also associated with a number of diseases that manifest with symptoms such as sensory ataxia, neuropathy, ophthalmoplegia, and hearing loss.

Subunit : Forms multimers in vitro, including hexamers. Interacts with POLG in vitro.

Subcellular Location : Mitochondrion matrix, mitochondrion nucleoid. Note=Colocalizes with mtDNA in mitochondrial nucleoids, a nucleoproteins complex consisting of a number of copies of proteins associated with mtDNA, probably involved in mtDNA maintenance and expression.

Tissue Specificity : High relative levels in skeletal muscle, testis and pancreas. Lower levels of expression in the heart, brain, placenta, lung, liver, kidney, spleen, thymus, prostate, ovary, small intestine, colon and leukocytes. Expression is coregulated with MRPL43.

DISEASE : Progressive external ophthalmoplegia with mitochondrial DNA deletions autosomal dominant 3 (PEOA3) [MIM:609286]: A disorder characterized by progressive weakness of ocular muscles and levator muscle of the upper eyelid. In a minority of cases, it is associated with skeletal myopathy, which predominantly involves axial or proximal muscles and which causes abnormal fatigability and even permanent muscle weakness. Ragged-red fibers and atrophy are found on muscle biopsy. A large proportion of chronic ophthalmoplegias are associated with other symptoms, leading to a multisystemic pattern of this disease. Additional symptoms are variable, and may include cataracts, hearing loss, sensory axonal neuropathy, ataxia, depression, hypogonadism, and parkinsonism. Note=The disease is caused by mutations affecting the gene represented in this entry. [DISEASE] Sensory ataxic neuropathy dysarthria and ophthalmoparesis (SANDO) [MIM:607459]: A systemic disorder resulting from mitochondrial dysfunction associated with mitochondrial depletion in skeletal muscle and peripheral nerve tissue. The clinical triad of symptoms consists of sensory ataxic neuropathy, dysarthria, and ophthalmoparesis. However, the phenotype varies widely, even within the same family, and can also include myopathy, seizures, and hearing loss. An atypical form of the disease is characterized by headaches and/or seizures manifesting in childhood or adolescence, followed by development of cerebellar and sensory ataxia, dysarthria, progressive external ophthalmoplegia, and myoclonus in early adulthood. Note=The disease is caused by mutations affecting the gene represented in this entry.

Mitochondrial DNA depletion syndrome 7 (MTDPS7) [MIM:271245]: A severe disease associated with mitochondrial dysfunction. Some patients are affected by progressive atrophy of the cerebellum, brain stem, the spinal cord, and sensory axonal neuropathy. Clinical features include hypotonia, athetosis, ataxia, ophthalmoplegia, sensorineural hearing deficit, sensory axonal neuropathy, epileptic encephalopathy and female hypogonadism. In some individuals liver dysfunction and multi-organ failure is present. Note=The disease is caused by mutations affecting the gene represented in this entry.

Similarity : Contains 1 SF4 helicase domain.

Database links : UniProtKB/Swiss-Prot: Q96RR1.1

抗体的鉴定：

1）蛋白8抗体费用 抗体的效价鉴定：不管是用于诊断还是用于,制备抗体的目的都是要求较高效价。不同的抗原制备的抗体,要求的效价不一。鉴定效价的方法很多,包括有试管凝集反应,琼脂扩散试验,酶联免疫吸附试验等。常用的抗原所制备的抗体一般都有约成的鉴定效价的方法,以资比较。如制备抗抗体的效价,一般就采用琼脂扩散试验来鉴定。

2）抗体的特异性鉴定：抗体的特异性是指与相应抗原或近似抗原物质的识别能力。抗体的特异性高,它的识别能力就强。衡量特异性通常以交叉反应率来表示。交叉反应率可用竞争抑制试验测定。以不同浓度抗原和近似抗原分别做竞争抑制曲线,计算各自的结合率,求出各自在IC50时的浓度,并按公式计算交叉反应率。 

如果所用抗原浓度IC50浓度为pg/管,而一些近似抗原物质的IC50浓度几乎是无穷大时,表示这一抗血清与其他抗原物质的交叉反应率近似为0,即该血清的特异性较好。

3）抗体亲和力：是指抗体和抗原结合的牢固程度。亲和力的高低是由抗原分子的大小,抗体分子的结合位点与抗原决定簇之间立体构型的合适度决定的。有助于维持抗原抗体复合物稳定的分子间力有氢键,疏水键,侧链相反电荷基因的库仑力,范德华力和空间斥力。亲和力常以亲和常数K表示,K的单位是L/mol。抗体亲和力的测定对抗体的筛选,确定抗体的用途,验证抗体的均一性等均有重要意义。

ER- Alpha(phospho-Tyr537)peptide 磷酸化雌激素受体α抗原Multi-class antibodies规格： 0.5mg

极光激酶B抗体 Anti-Aurora B 0.1ml

Rhesus antibody Rh phospho-EGFR (Thr693) 磷酸化表皮生长因子受体抗体 规格 0.1ml

D-MEM/F-12培养基(原装) 10×1L Gibco

ZNF532 英文名称： 锌指蛋白532抗体 0.2ml

DR3 英文名称： 死亡受体3抗体 0.1ml

极光激酶B抗体 Anti-Aurora B 0.1ml

ER- Alpha(phospho-Ser167)peptide 人磷酸化雌激素受体α多肽抗原Multi-class antibodies规格： 0.5mg

自分泌运动因子抗体 Anei-ATX/Autotaxin/E-NPP2 0.1ml

Rhesus antibody Rh phospho-EGFR (Tyr1016) 磷酸化表皮生长因子受体抗体 规格 0.1ml

Neurobasal Medium 500ml Gibco

ZNF516 英文名称： 锌指蛋白516抗体 0.2ml

DR4/ 英文名称： 死亡受体4抗体 0.1ml

自分泌运动因子抗体 Anei-ATX/Autotaxin/E-NPP2 0.1ml

DKK1(Human Dickkopf 1) ELISA Kit 人Dickkopf 1Multi-class antibodies规格： 48T

Anti-HCV-NS3 病毒-NS3抗体Multi-class antibodies规格： 0.1ml

Rhesus antibody Rh MIG6/ERRFI1 有丝分裂原诱导基因6抗体 规格 0.2ml

FAK(Human focal adhesion kinase) ELISA Kit 人黏着斑激酶 96T

ABCA4 英文名称： 神经元突触膜胞外分泌调节蛋白1抗体 0.2ml

CG028/C7orf28A 英文名称： 7号染色体开放阅读框28A抗体 0.1ml

Anti-HCV-NS3 病毒-NS3抗体Multi-class antibodies规格： 0.1ml

人补体片段3c(C3c)ELISA 试剂盒 96T/48T 试剂盒 组装/原装

豚鼠单核细胞趋化蛋白1(MCP-1/CCL2/MCAF)免疫试剂盒 Guinea pig monocyte chemotactic protein 1,MCP1 ELISA Kit

羧化基质谷酸蛋白(ucMGP)ELISA试剂盒 ，英文名： ucMGP ELISA Kit

MouseUbiquitin,UbELISA试剂盒小鼠泛素蛋白(Ub)ELISA试剂盒规格：96T/48T

Elisa猪瘟抗原2板检测试剂盒2*96孔2*96孔

HumanAngiotensinⅠConveingEnzyme,ACEⅠELISAKit人紧张素Ⅰ转化酶(ACEⅠ)ELISA试剂盒规格：96T/48T

大鼠透明质酸结合蛋白1(HABP1)ELISA试剂盒 ，英文名： HABP1 ELISA Kit

Bovine lymphocyte factor ELISA Kit 牛淋巴细胞因子ELISA试剂盒

Ratleoopichormone,LHELISAKit 大鼠促黄体激素(LH)ELISA试剂盒 96T/48T 进口分装

CLIAKitforHp-IgGELISAKit大鼠幽门螺旋菌IgG规格：48T/96T

细胞PHKG2激酶活性定量检测试剂盒(A/B/C)20次

ELISAKitTLR-9/CD289大鼠Toll样受体9规格：48T/96T

蛋白8抗体费用 大鼠纤维胶凝蛋白1(FCN1)ELISA试剂盒 ，英文名： FCN1 ELISA Kit

Mouse heat shock factor 1 (HSF1) ELISA Kit 小鼠热休克因子1(HSF1)ELISA试剂盒

MousesolubleE-selectin,sE-selectinELISAKit 小鼠可溶性E选择素(sE-selectin)ELISA试剂盒 96T/48T 进口分装

CLIAKitforHumanadvancedoxidationproteinproducts,AOPPELISAKit人晚期氧化蛋白产物规格：48T/96T

细胞HHV6-B(HUMANHERPESVIRUS6-B)病毒定性检测试剂盒20次

ELISAKitC5a大鼠补体片断5a规格：48T/96T