抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 細(xì)胞周期檢控Rad17蛋白抗體（復(fù)制因子C）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)不同的分子比來篩選最適條件；

3.用NHSB量過量也是不利的,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-Rad17/RFC 

中文名稱   細(xì)胞周期檢控Rad17蛋白抗體（復(fù)制因子C）費(fèi)用 

別    名  CCYC; Cell cycle checkpoint protein; Cell cycle checkpoint protein RAD17; HRAD 17; hRad17; R24L; Rad 17; Rad 24; RAD1 (S. pombe) homolog; RAD17; RAD17 homolog; Rad17 like protein; RAD17_HUMAN; RAD17Sp; Rad24; RF C activator 1 homolog; RF C/activator 1 homolog; RF-C/activator 1 homolog.

濃    度   1mg/1ml

規(guī) 格  0.1ml/100μg 0.2ml/200μg

抗體來源   Rabbit

克隆類型   polyclonal

交叉反應(yīng)   Human, Mouse, Rat, Dog, Pig, Cow, Horse, Rabbit 

產(chǎn)品類型   一抗  

研究領(lǐng)域    免疫學(xué) 染色質(zhì)和核信號(hào) 表觀遺傳學(xué)

蛋白分子量  predicted molecular weight: 77kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human Rad17 

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

細(xì)胞周期檢控Rad17蛋白抗體（復(fù)制因子C）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 ICC=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 The protein encoded by this gene is highly similar to the gene product of Schizosaccharomyces pombe rad17, a cell cycle checkpoint gene required for cell cycle arrest and DNA damage repair in response to DNA damage. This protein shares strong similarity with DNA replication factor C (RFC), and can form a complex with RFCs. This protein binds to chromatin prior to DNA damage and is phosphorylated by the checkpoint kinase ATR following damage. This protein recruits the RAD1-RAD9-HUS1 checkpoint protein complex onto chromatin after DNA damage, which may be required for its phosphorylation. The phosphorylation of this protein is required for the DNA-damage-induced cell cycle G2 arrest, and is thought to be a critical early event during checkpoint signaling in DNA-damaged cells. Multiple alternatively spliced transcript variants of this gene, which encode four distinct protein isoforms, have been reported. Two pseudogenes, located on chromosomes 7 and 13, have been identified. [provided by RefSeq, Jul 2013].

Function : Essential for sustained cell growth, maintenance of chromosomal stability, and ATR-dependent checkpoint activation upon DNA damage. Has a weak ATPase activity required for binding to chromatin. Participates in the recruitment of the RAD1-RAD9-HUS1 complex and C12orf32/RHINO onto chromatin, and in CHEK1 activation. May also serve as a sensor of DNA replication progression, and may be involved in homologous recombination.

Subunit : Part of a DNA-binding complex containing RFC2, RFC3, RFC4 and RFC5. Interacts with RAD1 and RAD9 within the RAD1-RAD9-HUS1 complex. Interacts with RAD9B, POLE, NHP2L1 and MCM7. DNA damage promotes interaction with ATR or ATM and disrupts interaction with the RAD1-RAD9-HUS1 complex.

Subcellular Location : Nucleus. Note=Phosphorylated form redistributes to discrete nuclear foci upon DNA damage.

Tissue Specificity : Overexpressed in various cancer cell lines and in colon carcinoma (at protein level). Isoform 2 and isoform 3 are the most abundant isoforms in non irradiated cells (at protein level). Ubiquitous at low levels. Highly expressed in testis, where it is expressed within the germinal epithelium of the seminiferous tubuli. Weakly expressed in seminomas (testicular tumors).

Post-translational modifications : Phosphorylated. Phosphorylation on Ser-646 and Ser-656 is cell cycle-regulated, enhanced by genotoxic stress, and required for activation of checkpoint signaling. Phosphorylation is mediated by ATR upon UV or replication arrest, whereas it may be mediated both by ATR and ATM upon ionizing radiation. Phosphorylation on both sites is required for interaction with RAD1 but dispensable for interaction with RFC3 or RFC4.

Similarity : Belongs to the rad17/RAD24 family.

Database links : UniProtKB/Swiss-Prot: O75943

抗體的鑒定：

1）細(xì)胞周期檢控Rad17蛋白抗體（復(fù)制因子C）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?？贵w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。

CD122/IL-2RB(Imterleukm-2 Receptor beta) CD122/白介素2受體β鏈抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-beta-Amyloid(1-42) β淀粉樣肽(1-42)抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh phospho-Bid (Ser65) 磷酸化BH3結(jié)構(gòu)域凋亡誘導(dǎo)蛋白抗體 規(guī)格 0.1ml

ELISA試劑盒開發(fā)服務(wù)(10kit以內(nèi)) 次 西唐

VWF 英文名稱： 假性因子/性因子抗體 0.1ml

DPF2 英文名稱： 雙PHD-D4鋅指蛋白2抗體 0.2ml

Anti-beta-Amyloid(1-42) β淀粉樣肽(1-42)抗體Multi-class antibodies規(guī)格： 0.1ml

Anti-Phospho-PNK1 (Ser114/Thr118) /FITC 熒光素標(biāo)記磷酸化多聚合苷酸激酶3磷酸化酶抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rabbit anti-mouse IgG1, kappa chain 兔抗小鼠IgG1k鏈 (親和層析純化)Multi-class antibodies規(guī)格： 1mg

葡萄糖調(diào)節(jié)蛋白94抗體 Anti-GRP94 0.1ml

Dog IgM/Cy5.5 Cy5.5標(biāo)記的兔抗犬IgM抗體 0.1ml

FAM160B1 英文名稱： FAM160B1蛋白抗體 0.2ml

Rhesus antibody Rh Phospho-S6 Ribosomal Protein (Ser235+Ser236) 磷酸化S6核糖體蛋白抗體 規(guī)格 0.1ml

Rabbit anti-mouse IgG1, kappa chain 兔抗小鼠IgG1k鏈 (親和層析純化)Multi-class antibodies規(guī)格： 1mg

SYNPR/Synaptoporin 英文名稱： 突觸囊泡蛋白抗體 0.2ml

EDG1 英文名稱： 內(nèi)皮細(xì)胞分化鞘脂G蛋白偶聯(lián)受體1抗體 0.1ml

CREB結(jié)合蛋白抗體 Anti-CBP 0.1ml

Anti-ULBP1/N2DL1/FITC 熒光素標(biāo)記兔抗人、大、小鼠UL16結(jié)合蛋白1蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh Phospho-Histone H3 (Ser28) 磷酸化組蛋白H3抗體 規(guī)格 0.1ml

VEGF(Vascular endothelial growth factor) 內(nèi)皮生長(zhǎng)因子(多肽抗原)Multi-class antibodies規(guī)格： 0.5mg

RLF, 大鼠淋巴成纖維細(xì)胞

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FAP Others Human 人 FAP / Seprase 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

CL-0290MDA-MB-468(人癌細(xì)胞)5×106cells/瓶×2

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CL-0154MDCK(犬腎細(xì)胞)5×106cells/瓶×2

IFNB1 Others Human 人 Ierferon beta / IFN-beta / IFNB CHO細(xì)胞裂解液 (陽(yáng)性對(duì)照)

人肝竇內(nèi)皮細(xì)胞裂解物HHSECL

小香豬皮膚細(xì)胞;SSC-S2 組織細(xì)胞細(xì)胞，U-937細(xì)胞 NUTU-19細(xì)胞，大鼠細(xì)胞株

SV40轉(zhuǎn)染人成骨細(xì)胞;hFOB 1.19

HA Others H1N3 甲型 H1N3 (A/duck/NZL/160/1976) 血凝素HA1 (Hemagglutinin) 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

細(xì)胞周期檢控Rad17蛋白抗體（復(fù)制因子C）費(fèi)用 RLF, 大鼠淋巴成纖維細(xì)胞

HEPG2.2.15 細(xì)胞HBV病毒株

FAP Others Human 人 FAP / Seprase 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

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