抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. piwi樣1蛋白抗體費(fèi)用 如在反應(yīng)混合液中有疊氮鈉或游離氨基存在,會(huì)抑制標(biāo)記反應(yīng)。因此,蛋白質(zhì)在反應(yīng)前要對(duì) 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(zhì)之間的分子比按蛋白質(zhì)表面的ε-氨基的密度會(huì)有所不同,選擇不當(dāng)則影響標(biāo)記的效率,應(yīng)先用幾個(gè)不同的分子比來(lái)篩選最適條件；

3.用NHSB量過(guò)量也是不利的,抗原的結(jié)合位點(diǎn)可能因此被封閉,導(dǎo)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(shí)可加入去污劑如 Triton x-100, Tween20等；

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

6.生物素還可能與不同的功能基團(tuán),如羰基、氨基、巰基、異咪唑基及苯酚基,也可與糖基共價(jià)結(jié)合；

7.交聯(lián)反應(yīng)后,應(yīng)充分透析,否則,殘余的生物素會(huì)對(duì)生物素化抗體與親和素的結(jié)合產(chǎn)生競(jìng)爭(zhēng)作用；

8.在細(xì)胞的熒光標(biāo)記實(shí)驗(yàn)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(duì)某些細(xì)胞可導(dǎo)致高本底。

產(chǎn)品訂購(gòu)信息：
英文名稱  Anti-PIWIL1/Miwi 

中文名稱   piwi樣1蛋白抗體費(fèi)用 

別    名  HIWI; MIWI; P-element induced wimpy testis; Piwi; Piwi homolog; Piwi like 1; Piwi like protein 1; PIWIL 1; PIWIL1; HIWI; MIWI; Piwi (Drosophila) like 1; PIWI; Piwi homolog; Piwi like 1 (drosophila); Piwi like 1; Piwi-like protein 1; PIWIL 1; PIWIL1; PIWL1; piwi-like protein 1 isoform 1; PIWL1_HUMAN.

濃    度   1mg/1ml

規(guī) 格  0.1ml/100μg 0.2ml/200μg

抗體來(lái)源   Rabbit

克隆類型   polyclonal

交叉反應(yīng)   Human, Mouse, Rat, Chicken 

產(chǎn)品類型   一抗  

研究領(lǐng)域     染色質(zhì)和核信號(hào) 信號(hào)轉(zhuǎn)導(dǎo) 干細(xì)胞 轉(zhuǎn)錄調(diào)節(jié)因子

蛋白分子量  predicted molecular weight: 95kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human PIWIL1 C-terminus 

亞    型   IgG

純化方法   affinity purified by Protein A

儲(chǔ) 存 液   0.01M PBS, pH 7.4 with 10 mg/ml BSA and 0.1% Sodium azide

piwi樣1蛋白抗體費(fèi)用 產(chǎn)品應(yīng)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

（石蠟切片需做抗原修復(fù)） 

 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 This gene encodes a member of the PIWI subfamily of Argonaute proteins, evolutionarily conserved proteins containing both PAZ and Piwi motifs that play important roles in stem cell self-renewal, RNA silencing, and translational regulation in diverse organisms. The encoded protein may play a role as an intrinsic regulator of the self-renewal capacity of germline and hematopoietic stem cells. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jul 2010].

Function : Plays a central role during spermatogenesis by repressing transposable elements and prevent their mobilization, which is essential for the germline integrity. Acts via the piRNA metabolic process, which mediates the repression of transposable elements during meiosis by forming complexes composed of piRNAs and Piwi proteins and govern the methylation and subsequent repression of transposons. Directly binds methylated piRNAs, a class of 24 to 30 nucleotide RNAs that are generated by a Dicer-independent mechanism and are primarily derived from transposons and other repeated sequence elements. Besides their function in transposable elements repression, piRNAs are probably involved in other processes during meiosis such as translation regulation. Probable component of some RISC complex, which mediates RNA cleavage and translational silencing. Also plays a role in the formation of chromatoid bodies and is required for some miRNAs stability. Isoform 3 may be a negative developmental regulator.

Subunit : Interacts (via Piwi domain) with DICER1, suggesting that it forms ribonucleoprotein RISC complexes. This interaction is regulated by HSP90AB1 activity. Interacts with MAEL, KIF17, PABPC1, PRMT5 and WDR77. Interacts (when methylated on arginine residues) with TDRD1, TDRKH/TDRD2, RNF17/TDRD4, TDRD6, TDRD7 and TDRD9.

Subcellular Location : Cytoplasm. Note=Component of the meiotic nuage, also named P granule, a germ-cell-specific organelle required to repress transposon during meiosis. Also present in chromatoid body.

Post-translational modifications : Arginine methylation by PRMT5 is required for the interaction with Tudor domain-containing protein (TDRD1, TDRKH/TDRD2, RNF17/TDRD4, TDRD6, TDRD7 and TDRD9) and subsequent localization to the meiotic nuage, also named P granule.

Similarity : Belongs to the argonaute family. Piwi subfamily.

Contains 1 PAZ domain.

Contains 1 Piwi domain.

Database links :

NCBI Reference Sequence: NP_004755

UniProtKB/Swiss-Prot: Q96J94

Entrez Gene: 9271 Human

Entrez Gene: 57749 Mouse

Entrez Gene: 363912 Rat

Omim: 605571 Human

SwissProt: Q96J94 Human

SwissProt: Q9JMB7 Mouse

Unigene: 405659 Human

Unigene: 272720 Mouse

抗體的鑒定：

1）piwi樣1蛋白抗體費(fèi)用 抗體的效價(jià)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(jià)。不同的抗原制備的抗體,要求的效價(jià)不一。鑒定效價(jià)的方法很多,包括有試管凝集反應(yīng),瓊脂擴(kuò)散試驗(yàn),酶聯(lián)免疫吸附試驗(yàn)等。常用的抗原所制備的抗體一般都有約成的鑒定效價(jià)的方法,以資比較。如制備抗抗體的效價(jià),一般就采用瓊脂擴(kuò)散試驗(yàn)來(lái)鑒定。

2）抗體的特異性鑒定：抗體的特異性是指與相應(yīng)抗原或近似抗原物質(zhì)的識(shí)別能力?？贵w的特異性高,它的識(shí)別能力就強(qiáng)。衡量特異性通常以交叉反應(yīng)率來(lái)表示。交叉反應(yīng)率可用競(jìng)爭(zhēng)抑制試驗(yàn)測(cè)定。以不同濃度抗原和近似抗原分別做競(jìng)爭(zhēng)抑制曲線,計(jì)算各自的結(jié)合率,求出各自在IC50時(shí)的濃度,并按公式計(jì)算交叉反應(yīng)率。 

如果所用抗原濃度IC50濃度為pg/管,而一些近似抗原物質(zhì)的IC50濃度幾乎是無(wú)窮大時(shí),表示這一抗血清與其他抗原物質(zhì)的交叉反應(yīng)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(jié)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(jié)合位點(diǎn)與抗原決定簇之間立體構(gòu)型的合適度決定的。有助于維持抗原抗體復(fù)合物穩(wěn)定的分子間力有氫鍵,疏水鍵,側(cè)鏈相反電荷基因的庫(kù)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。抗體親和力的測(cè)定對(duì)抗體的篩選,確定抗體的用途,驗(yàn)證抗體的均一性等均有重要意義。

Anti-B7-1/CD80 /FITC 熒光素標(biāo)記抗刺激分子B7-1蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-IGF-1 R/FITC 熒光素標(biāo)記樣生長(zhǎng)因子-I受體抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh CD45/B220 白細(xì)胞共同抗原抗體 規(guī)格 0.1ml

NR1/NMDAR1 谷酸受體1 0.5mg

human Serum IgA 英文名稱： 人血清型免疫球蛋白A抗體 0.2ml

Rhesus antibody Rh Syndecan 2/heparin sulphate protoglycans 1 肝素糖蛋白1抗體抗體 規(guī)格 0.2ml

Anti-IGF-1 R/FITC 熒光素標(biāo)記樣生長(zhǎng)因子-I受體抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-HisX6/6His/His tag(CT)/HRP 辣根過(guò)氧化物酶標(biāo)記聚組氨酸抗體/抗His tag標(biāo)簽抗體(C端)IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-phospho-STAT1(pThr701)/FITC 熒光素標(biāo)記抗磷酸化信號(hào)轉(zhuǎn)導(dǎo)與轉(zhuǎn)錄激活因子1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh ATG3 自噬相關(guān)蛋白3抗體 規(guī)格 0.1ml

Beta-Amyloid (31-35) β淀粉樣肽31-35(多肽蛋白) 0.1mg

phospho-GIT1(Tyr554) 英文名稱： 磷酸化G蛋白偶聯(lián)受體激酶相互作用蛋白1 0.1ml

Rhesus antibody Rh Rabbit Anti-mouse IgM 兔抗小鼠IgM 規(guī)格 1mg

Anti-phospho-STAT1(pThr701)/FITC 熒光素標(biāo)記抗磷酸化信號(hào)轉(zhuǎn)導(dǎo)與轉(zhuǎn)錄激活因子1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

CDK5(Human Cyclin Dependent Kinase 5) ELISA Kit 人周期素依賴性激酶5Multi-class antibodies規(guī)格： 48T

Anti-Nck1/Nck2 NCK銜接因子蛋白2抗體Multi-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh IL-6 白介素6抗體 規(guī)格 0.1ml

Beta2-GP1 IgA/G/M(Human Beta2-glycoprotein 1 antibody IgA/G/M) ELISA Kit 人β2糖蛋白1抗體IgA/G/M 96T

phospho-PAK2 (Ser141) 英文名稱： 磷酸化p21激活激酶2抗體 0.1ml

CD44v10 英文名稱： CD44V10抗體 0.1ml

Anti-Nck1/Nck2 NCK銜接因子蛋白2抗體Multi-class antibodies規(guī)格： 0.2ml

CL-0234TM3(小鼠間質(zhì)細(xì)胞)5×106cells/瓶×2

KIT Others Human 人 KIT / c-KIT / CD117 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

人肺動(dòng)脈內(nèi)皮細(xì)胞cDNAHPAEC cDNA

小鼠髖動(dòng)脈內(nèi)皮細(xì)胞;PIEC 細(xì)胞，RBL-2H3細(xì)胞 ME細(xì)胞，C57小鼠瘤株

Sars結(jié)構(gòu)蛋白表達(dá)株;293 001B

HA Others H1N1 甲型 H1N1 (A/Puerto Rico/8/1934) 血凝素 (Hemagglutinin / HA) 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

CL-0232TF-1(人血液細(xì)胞)5×106cells/瓶×2

PTK6 Others Human 人 PTK6 / Brk 桿狀病毒-昆蟲細(xì)胞裂解液 (陽(yáng)性對(duì)照)

人平滑肌細(xì)胞cDNAHCSMC cDNA

NCL-H460細(xì)胞，非小細(xì)胞 人肺動(dòng)脈內(nèi)皮細(xì)胞,HPAEC細(xì)胞 肝動(dòng)脈平滑肌細(xì)胞Many types of cells包裝：5 × 105次方(1ml)

犬腎細(xì)胞;MDCK(NBL-2)

TNFRSF10A Others Human 人 TNFRSF10A / CD261 / APO2 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

piwi樣1蛋白抗體費(fèi)用 HA Others H1N1 甲型 H1N1 (A/Beijing/22808/2009) 血凝素 (Hemagglutinin / HA) 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

CM-H044人肝動(dòng)脈內(nèi)皮細(xì)胞完全培養(yǎng)基100mL

小鼠牙細(xì)胞完全培養(yǎng)基 100mL

NCI-H929-CMV-EGFP(穩(wěn)定株)人細(xì)胞 NCI-H929-CMV-EGFP (stable sain) human myeloma cell 1640+20% FBS(熱滅活)

IL15 Protein Human 重組人 IL-15 / IL15 / Ierleukin 15 蛋白 (His 標(biāo)簽)

人小神經(jīng)膠質(zhì)細(xì)胞(HM) ( 5×105 ) 人海馬神經(jīng)元 Human