抗體的生物素化標記實驗要點：

1. 蛋白8抗體費用 如在反應混合液中有疊氮鈉或游離氨基存在,會抑制標記反應。因此,蛋白質在反應前要對 0.1mol/L碳酸氫鈉緩沖液或0.5mol/L硼酸緩沖液充分透析；

2.所用的NHSB及待生物素化蛋白質之間的分子比按蛋白質表面的ε-氨基的密度會有所不同,選擇不當則影響標記的效率,應先用幾個不同的分子比來篩選最適條件；

3.用NHSB量過量也是不利的,抗原的結合位點可能因此被封閉,導致抗體失活；

4.由于抗體的氨基不易接近可能造成生物素化不足,此時可加入去污劑如 Triton x-100, Tween20等；

5.當游離ε-氨基(賴氨酸殘基的氨基)存在于抗體的抗原結合位點時,或位于酶的催化位點時,生物素化會降低或損傷抗體蛋白的結合力或活性；

6.生物素還可能與不同的功能基團,如羰基、氨基、巰基、異咪唑基及苯酚基,也可與糖基共價結合；

7.交聯(lián)反應后,應充分透析,否則,殘余的生物素會對生物素化抗體與親和素的結合產(chǎn)生競爭作用；

8.在細胞的熒光標記實驗中,中和親和素的本底低,但由于鏈霉親和素含有少量正電荷,故對某些細胞可導致高本底。

產(chǎn)品訂購信息：
英文名稱  Anti-Twinkle/ATXN8 

中文名稱   蛋白8抗體費用 

別    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.

濃    度   1mg/1ml

規(guī) 格  0.2ml/200μg

抗體來源   Rabbit

克隆類型   polyclonal

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

產(chǎn)品類型   一抗  

研究領域    細胞生物 神經(jīng)生物學 線粒體 表觀遺傳學

蛋白分子量  predicted molecular weight: 77kDa

性    狀   Lyophilized or Liquid

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

亞    型   IgG

純化方法   affinity purified by Protein A

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

蛋白8抗體費用 產(chǎn)品應用    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

（石蠟切片需做抗原修復） 

 not yet tested in other applications.

 optimal dilutions/concentrations should be determined by the end user.  

保存條件  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. 

產(chǎn)品介紹 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

抗體的鑒定：

1）蛋白8抗體費用 抗體的效價鑒定：不管是用于診斷還是用于,制備抗體的目的都是要求較高效價。不同的抗原制備的抗體,要求的效價不一。鑒定效價的方法很多,包括有試管凝集反應,瓊脂擴散試驗,酶聯(lián)免疫吸附試驗等。常用的抗原所制備的抗體一般都有約成的鑒定效價的方法,以資比較。如制備抗抗體的效價,一般就采用瓊脂擴散試驗來鑒定。

2）抗體的特異性鑒定：抗體的特異性是指與相應抗原或近似抗原物質的識別能力?？贵w的特異性高,它的識別能力就強。衡量特異性通常以交叉反應率來表示。交叉反應率可用競爭抑制試驗測定。以不同濃度抗原和近似抗原分別做競爭抑制曲線,計算各自的結合率,求出各自在IC50時的濃度,并按公式計算交叉反應率。 

如果所用抗原濃度IC50濃度為pg/管,而一些近似抗原物質的IC50濃度幾乎是無窮大時,表示這一抗血清與其他抗原物質的交叉反應率近似為0,即該血清的特異性較好。

3）抗體親和力：是指抗體和抗原結合的牢固程度。親和力的高低是由抗原分子的大小,抗體分子的結合位點與抗原決定簇之間立體構型的合適度決定的。有助于維持抗原抗體復合物穩(wěn)定的分子間力有氫鍵,疏水鍵,側鏈相反電荷基因的庫侖力,范德華力和空間斥力。親和力常以親和常數(shù)K表示,K的單位是L/mol?？贵w親和力的測定對抗體的篩選,確定抗體的用途,驗證抗體的均一性等均有重要意義。

ER- Alpha(phospho-Tyr537)peptide 磷酸化雌激素受體α抗原Multi-class antibodies規(guī)格： 0.5mg

極光激酶B抗體 Anti-Aurora B 0.1ml

Rhesus antibody Rh phospho-EGFR (Thr693) 磷酸化表皮生長因子受體抗體 規(guī)格 0.1ml

D-MEM/F-12培養(yǎng)基(原裝) 10×1L Gibco

ZNF532 英文名稱： 鋅指蛋白532抗體 0.2ml

DR3 英文名稱： 死亡受體3抗體 0.1ml

極光激酶B抗體 Anti-Aurora B 0.1ml

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

自分泌運動因子抗體 Anei-ATX/Autotaxin/E-NPP2 0.1ml

Rhesus antibody Rh phospho-EGFR (Tyr1016) 磷酸化表皮生長因子受體抗體 規(guī)格 0.1ml

Neurobasal Medium 500ml Gibco

ZNF516 英文名稱： 鋅指蛋白516抗體 0.2ml

DR4/ 英文名稱： 死亡受體4抗體 0.1ml

自分泌運動因子抗體 Anei-ATX/Autotaxin/E-NPP2 0.1ml

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

Anti-HCV-NS3 病毒-NS3抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh MIG6/ERRFI1 有絲分裂原誘導基因6抗體 規(guī)格 0.2ml

FAK(Human focal adhesion kinase) ELISA Kit 人黏著斑激酶 96T

ABCA4 英文名稱： 神經(jīng)元突觸膜胞外分泌調節(jié)蛋白1抗體 0.2ml

CG028/C7orf28A 英文名稱： 7號染色體開放閱讀框28A抗體 0.1ml

Anti-HCV-NS3 病毒-NS3抗體Multi-class antibodies規(guī)格： 0.1ml

人補體片段3c(C3c)ELISA 試劑盒 96T/48T 試劑盒 組裝/原裝

豚鼠單核細胞趨化蛋白1(MCP-1/CCL2/MCAF)免疫試劑盒 Guinea pig monocyte chemotactic protein 1,MCP1 ELISA Kit

羧化基質谷酸蛋白(ucMGP)ELISA試劑盒 ，英文名： ucMGP ELISA Kit

MouseUbiquitin,UbELISA試劑盒小鼠泛素蛋白(Ub)ELISA試劑盒規(guī)格：96T/48T

Elisa豬瘟抗原2板檢測試劑盒2*96孔2*96孔

HumanAngiotensinⅠConveingEnzyme,ACEⅠELISAKit人緊張素Ⅰ轉化酶(ACEⅠ)ELISA試劑盒規(guī)格：96T/48T

大鼠透明質酸結合蛋白1(HABP1)ELISA試劑盒 ，英文名： HABP1 ELISA Kit

Bovine lymphocyte factor ELISA Kit 牛淋巴細胞因子ELISA試劑盒

Ratleoopichormone,LHELISAKit 大鼠促黃體激素(LH)ELISA試劑盒 96T/48T 進口分裝

CLIAKitforHp-IgGELISAKit大鼠幽門螺旋菌IgG規(guī)格：48T/96T

細胞PHKG2激酶活性定量檢測試劑盒(A/B/C)20次

ELISAKitTLR-9/CD289大鼠Toll樣受體9規(guī)格：48T/96T

蛋白8抗體費用 大鼠纖維膠凝蛋白1(FCN1)ELISA試劑盒 ，英文名： FCN1 ELISA Kit

Mouse heat shock factor 1 (HSF1) ELISA Kit 小鼠熱休克因子1(HSF1)ELISA試劑盒

MousesolubleE-selectin,sE-selectinELISAKit 小鼠可溶性E選擇素(sE-selectin)ELISA試劑盒 96T/48T 進口分裝

CLIAKitforHumanadvancedoxidationproteinproducts,AOPPELISAKit人晚期氧化蛋白產(chǎn)物規(guī)格：48T/96T

細胞HHV6-B(HUMANHERPESVIRUS6-B)病毒定性檢測試劑盒20次

ELISAKitC5a大鼠補體片斷5a規(guī)格：48T/96T